BACKGROUND The pyruvate dehydrogenase E1 subunitβ(PDHB)gene which regulates energy metabolism is located in mitochondria.However,few studies have elucidated the role and mechanism of PDHB in different cancers.AIM To ...BACKGROUND The pyruvate dehydrogenase E1 subunitβ(PDHB)gene which regulates energy metabolism is located in mitochondria.However,few studies have elucidated the role and mechanism of PDHB in different cancers.AIM To comprehensive pan-cancer analysis of PDHB was performed based on bioinformatics approaches to explore its tumor diagnostic and prognostic value and tumor immune relevance in cancer.In vitro experiments were performed to examine the biological regulation of PDHB in liver cancer.METHODS Pan-cancer data related to PDHB were obtained from the Cancer Genome Atlas(TCGA)database.Analysis of the gene expression profiles of PDHB was based on TCGA and Genotype Tissue Expression Dataset databases.Cox regression analysis and Kaplan-Meier methods were used to assess the correlation between PDHB expression and survival prognosis in cancer patients.The correlation between PDHB and receiver operating characteristic diagnostic curve,clinicopathological staging,somatic mutation,tumor mutation burden(TMB),microsatellite instability(MSI),DNA methylation,and drug susceptibility in pan-cancer was also analyzed.Various algorithms were used to analyze the correlation between PDHB and immune cell infiltration and tumor chemotaxis environment,as well as the co-expression analysis of PDHB and immune checkpoint(ICP)genes.The expression and functional phenotype of PDHB in single tumor cells were studied by single-cell sequencing,and the functional enrichment analysis of PDHB-related genes was performed.The study also validated the level of mRNA or protein expression of PDHB in several cancers.Finally,in vitro experiments verified the regulatory effect of PDHB on the proliferation,migration,and invasion of liver cancer.RESULTS PDHB was significantly and differently expressed in most cancers.PDHB was significantly associated with prognosis in patients with a wide range of cancers,including kidney renal clear cell carcinoma,kidney renal papillary cell carcinoma,breast invasive carcinoma,and brain lower grade glioma.In some cancers,PDHB expression was clearly associated with gene mutations,clinicopathological stages,and expression of TMB,MSI,and ICP genes.The expression of PDHB was closely related to the infiltration of multiple immune cells in the immune microenvironment and the regulation of tumor chemotaxis environment.In addition,single-cell sequencing results showed that PDHB correlated with different biological phenotypes of multiple cancer single cells.This study further demonstrated that down-regulation of PDHB expression inhibited the proliferation,migration,and invasion functions of hepatoma cells.CONCLUSION As a member of pan-cancer,PDHB may be a novel cancer marker with potential value in diagnosing cancer,predicting prognosis,and in targeted therapy.展开更多
Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate can...Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate cancer(PCa).In the present study,we aimed to explore the function of DLC1 in PCa cells.Methods:Silencing and overexpression of DLC1 were induced in an androgen-sensitive PCa cell line(LNCaP)using RNA interference and lentiviral vector transduction.The Cell Counting Kit-8 assay was performed to determine cell proliferation.The cell cycle was examined by performing a propidium iodide staining assay.Results:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of LNCaP cells.Moreover,DLC1 expression was negatively correlated with Rho-associated protein kinase(ROCK)expression in LNCaP cells.Importantly,this study showed that the ROCK inhibitor Y27632 restored the function of DLC1 in LNCaP cells and reduced the tumorigenicity of LNCaP cells in vivo.Conclusion:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of PCa cells and negatively correlated with ROCK expression in PCa cells and tissue.展开更多
目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值...目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值。方法选取2021年7月—2023年7月在玉林市第一人民医院就诊HBV病毒感染相关的50例肝癌患者,50例肝硬化患者,50例慢性乙型肝炎患者以及50名同期健康体检者作为研究对象。比较4组研究对象血清中CHI3L1、AFP、GGT、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)等水平的差异,用受试者工作特征(receiver operating characteristic curve,ROC)曲线评估各指标在肝癌中的诊断价值。结果肝炎组、肝硬化组、肝癌组的CHI3L1、AST、ALT水平均高于对照组,肝癌组AFP、GGT水平高于对照组,差异有统计学意义(P<0.05)。与肝炎组比较,肝硬化组及肝癌组的CHI3L1、AFP、GGT、AST、ALT水平均升高,差异有统计学意义(P<0.05)。与肝硬化组比较,肝癌组的CHI3L1、AFP、GGT、AST水平均升高,差异有统计学意义(P<0.05)。ROC曲线分析显示,CHI3L1、AFP、GGT联合时的曲线下面积(area under the curve,AUC)最大(AUC=0.936)。Spearman相关分析结果显示,CHI3L1与AST呈正相关(r=0.414,P=0.003),AFP与GGT呈正相关(r=0.437,P=0.002),AFP与AST呈正相关(r=0.504,P<0.001),GGT与AST呈正相关(r=0.759,P<0.001),GGT与ALT呈正相关(r=0.636,P<0.001)。结论CHI3L1、AFP及GGT联合检测可提高肝癌的诊断价值,对临床肝癌患者诊疗有重要作用。展开更多
Chronic alcohol consumption is a major risk factor worldwide affecting significantly both mortality and years of life lost （YLL） （1）. Ca. 5% of the western world show risky alcohol consumption and in some countrie...Chronic alcohol consumption is a major risk factor worldwide affecting significantly both mortality and years of life lost （YLL） （1）. Ca. 5% of the western world show risky alcohol consumption and in some countries such as China a regional yearly increase of alcohol consumption of over 400% has been observed recently （2,3）. The liver is the major target organ of alcohol. According to the recently published ＇Global Burden of Disease Study 2010, liver cirrhosis and liver cancer are ranked at position 12 and 16 in the global deaths statistics （1）. Thus, in 2010, ca. 1 million people died from liver cirrhosis with one third directly attributable to alcohol. This is a considerable number when comparing with coronary heart disease with 7 million deaths and the leading cause of mortality. In central Europe, liver cirrhosis even ranks at the fourth position in YLL. Hepatocellular cancer （HCC） is now the most common fatal complication of patients with alcoholic cirrhosis. Moreover, it shows the second fastest increase of all tumors worldwide after kidney tumors and alcohol-associated HCC ranks on third position after HCCs caused by viral hepatitis B and C.展开更多
Background:Hepatocellular carcinoma(HCC),the most common type of primary liver cancer,is the fourth leading cause of cancer-related deaths worldwide.Previous evidence shows that the expression of circulating RNA ZFR(c...Background:Hepatocellular carcinoma(HCC),the most common type of primary liver cancer,is the fourth leading cause of cancer-related deaths worldwide.Previous evidence shows that the expression of circulating RNA ZFR(circZFR)is upregulated in HCC tissues.However,the molecular mechanism of circZFR in HCC is unclear.Methods:Quantitative reverse transcriptase polymerase chain reaction(qRT-PCR)was employed to detect the expression of circZFR,microRNA-624-3p(miR-624-3p)and WEE1 in HCC tissues and cells.RNase R assay and actinomycin D treatment assay were used to analyze the characteristics of circZFR.For functional analysis,the capacities of colony formation,cell proliferation,cell apoptosis,migration and invasion were assessed by colony formation assay,5-ethynyl-2-deoxyuridine(EdU)assay,flow cytometry assay and transwell assay.Western blot was used to examine the protein levels of WEE1 and epithelial-mesenchymal transition(EMT)-related proteins.The interactions between miR-624-3p and circZFR or WEE1 were validated by dual-luciferase reporter assay and RNA immunoprecipitation(RIP)assay.Xenograft models were established to determine the role of circZFR in vivo.Results:circZFR and WEE1 were upregulated,while miR-624-3p expression was reduced in HCC tissues and cells.circZFR could sponge miR-624-3p,and WEE1 was a downstream gene of miR-624-3p.Knockdown of circZFR significantly reduced the malignant behaviors of HCC and that co-transfection with miR624-3p inhibitor restored this change.Overexpression of WEE1 abolished the inhibitory effect of miR624-3p mimic on HCC cells.Mechanistically,circZFR acted as a competitive endogenous RNA(ceRNA)to regulate WEE1 expression by targeting miR-624-3p.Furthermore,in vivo studies have illustrated that circZFR knockdown inhibited tumor growth.Conclusions:circZFR knockdown reduced HCC cell proliferation,migration and invasion and promoted apoptosis by regulating the miR-624-3p/WEE1 axis,suggesting that the circZFR/miR-624-3p/WEE1 axis might be a potential target for HCC treatment.展开更多
BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),t...BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),the mechanisms are largely unknown.AIM To explore the role of BMI-1,a polycomb group protein family member,which is highly expressed in LM,and the interaction between aHSCs and CRC cells in promoting CRC liver metastasis(CRLM).METHODS Immunohistochemistry was carried out to examine BMI-1 expression in LM and matched liver specimens of CRC.The expression levels of BMI-1 in mouse liver during CRLM(0,7,14,21,and 28 d)were detected by Western blotting(WB)and the quantitative polymerase chain reaction(qPCR)assay.We overexpressed BMI-1 in HSCs(LX2)by lentivirus infection and tested the molecular markers of aHSCs by WB,qPCR,and the immunofluorescence assay.CRC cells(HCT116 and DLD1)were cultured in HSC-conditioned medium(LX2 NC CM or LX2 BMI-1 CM).CM-induced CRC cell proliferation,migration,epithelial-mesenchymal transition(EMT)phenotype,and transforming growth factor beta(TGF-β)/SMAD pathway changes were investigated in vitro.A mouse subcutaneous xenotransplantation tumor model was established by co-implantation of HSCs(LX2 NC or LX2 BMI-1)and CRC cells to investigate the effects of HSCs on tumor growth and the EMT phenotype in vivo.RESULTS Positive of BMI-1 expression in the liver of CRLM patients was 77.8%.The expression level of BMI-1 continued to increase during CRLM in mouse liver cells.LX2 overexpressed BMI-1 was activated,accompanied by increased expression level of alpha smooth muscle actin,fibronectin,TGF-β1,matrix metalloproteinases,and interleukin 6.CRC cells cultured in BMI-1 CM exhibited enhanced proliferation and migration ability,EMT phenotype and activation of the TGF-β/SMAD pathway.In addition,the TGF-βR inhibitor SB-505124 diminished the effect of BMI-1 CM on SMAD2/3 phosphorylation in CRC cells.Furthermore,BMI-1 overexpressed LX2 HSCs promoted tumor growth and the EMT phenotype in vivo.CONCLUSION High expression of BMI-1 in liver cells is associated with CRLM progression.BMI-1 activates HSCs to secrete factors to form a prometastatic environment in the liver,and aHSCs promote proliferation,migration,and the EMT in CRC cells partially through the TGF-β/SMAD pathway.展开更多
AIM: To investigate the expression characteristics of peroxiredoxin 1(PRDX1) m RNA and protein in liver cancer cell lines and tissues.METHODS: The RNA sequencing data from 374 patients with liver cancer were obtained ...AIM: To investigate the expression characteristics of peroxiredoxin 1(PRDX1) m RNA and protein in liver cancer cell lines and tissues.METHODS: The RNA sequencing data from 374 patients with liver cancer were obtained from The Cancer Genome Atlas. The expression and clinical characteristics of PRDX1 m RNA were analyzed in this dataset. The Kaplan-Meier and Cox regression survival analysis was performed to determine the relationship between PRDX1 levels and patient survival. Subcellular fractionation and Western blotting were used to demonstrate the expression of PRDX1 protein in six liver cancer cell lines and 29 paired fresh tissue specimens. After bioinformatics prediction,a putative posttranslational modification form of PRDX1 was observed using immunofluorescence under confocal microscopy and immunoprecipitation analysis in liver cancer cells.RESULTS: The m RNA of PRDX1 gene was upregulated about 1.3-fold in tumor tissue compared with the adjacent non-tumor control(P = 0.005). Its abundance was significantly higher in men than women(P < 0.001). High levels of PRDX1 m RNA were associated with a shorter overall survival time(P =0.04) but not with recurrence-free survival. The Cox regression analysis demonstrated that patients with high PRDX1 m RNA showed about 1.9-fold increase of risk for death(P = 0.03). In liver cancer cells,PRDX1 protein was strongly expressed with multiple different bands. PRDX1 in the cytosol fraction existed near the theoretical molecular weight,whereas two higher molecular weight bands were present in the membrane/organelle and nuclear fractions. Importantly,the theoretical PRDX1 band was increased,whereas the high molecular weight form was decreased in tumor tissues. Subsequent experiments revealed that the high molecular weight bands of PRDX1 might result from the post-translational modification by small ubiquitin-like modifier-1(SUMO1).CONCLUSION: PRDX1 was overexpressed in the tumor tissues of liver cancer and served as an independent poor prognostic factor for overall survival. PRDX1 can be modified by SUMO to play specific roles in hepatocarcinogenesis.展开更多
BACKGROUND:Phospholipid scramblase 1(PLSCR1) not only participates in the transbilayer movement of phospholipids,but also plays a role in the pathogenesis and progression of cancers.The present study aimed to evaluate...BACKGROUND:Phospholipid scramblase 1(PLSCR1) not only participates in the transbilayer movement of phospholipids,but also plays a role in the pathogenesis and progression of cancers.The present study aimed to evaluate the effect of silencing PLSCR1 expression by RNA interference in colorectal cancer(CRC) and metastatic liver cancer.METHODS:The expression of PLSCR1 in CRC and metastatic liver cancer samples was assessed by immunohistochemistry.The cultured cells with the highest expression were selected for subsequent experiments.We designed three siRNA oligonucleotide segments targeted at PLSCR1.Successful transfection was confirmed.The biological behavior of the cells in proliferation,adhesion,migration and invasion was determined.RESULTS:PLSCR1 protein expression increased significantly in the majority of CRC and metastatic liver cancer samples compared with normal samples.Lovo cells had the highest expression of PLSCR1.The siRNA-390 oligonucleotide segment had the best silencing effect.After transfection,Lovo cell proliferation was significantly inhibited compared with the controls in the MTT assay.Laminin and fibronectin adhesion assays showed Lovo cell adhesion was also significantly inhibited.In the migration assay,the number of migrating cells in the PLSCR1 siRNA-390 group was 50±12,significantly lower than the number in the siRNA-N group(115±28) and in the control group(118±31).In an invasion test,the number of invading cells in the PLSCR1 siRNA-390 group was 60±18,significantly lower than that in the siRNA-N group(97±26) and the control group(103±24).CONCLUSIONS:PLSCR1 is overexpressed in CRC and metastatic liver cancer.Silencing of PLSCR1 by siRNA inhibits the proliferation,adhesion,migration and invasion of Lovo cells,which suggests that PLSCR1 contributes to the tumorigenesis and tumor progression of CRC.PLSCR1 may be a potential gene therapy target for CRC and associated metastatic liver cancer.展开更多
Summary： Poly （ADP-ribose） polymerase-1 （PARP-1） inhibitors and histone deacetylase （HDAC） in- hibitors have recently emerged as promising anticancer drugs. The aim of this study was to investigate the effect o...Summary： Poly （ADP-ribose） polymerase-1 （PARP-1） inhibitors and histone deacetylase （HDAC） in- hibitors have recently emerged as promising anticancer drugs. The aim of this study was to investigate the effect of combination treatment with the PARP inhibitor P J34 and HDAC inhibitor SAHA on the proliferation of liver cancer cells. Cell proliferation and apoptosis were assessed in three human liver cancer cell lines （HepG2, Hep3B and HCC-LM3） treated with PJ34 （8 μmol/L） and SAHA （1 panol/L）, alone or combined, by Cell Counting Kit-8 assay and flow cytometry, respectively. The nude mice bear- ing subcutaneous HepG2 tumors were administered different groups of drugs （10 mg/kg PJ34, 25 mg/kg SAHA, 10 mg/kg PJ34＋25 mg/kg SAHA）, and the inhibition rates of tumor growth were compared between groups. The results showed that combined use of P J34 and SAHA could synergistically inhibit the proliferation of liver cancer cell lines HepG2, Hep3B and HCC-LM3. The apoptosis rate of HepG2 cells treated with PJ34＋SAHA was significantly higher than that of HepG2 cells treated with PJ34 or SAHA alone （P〈0.05）. In vivo, the tumor inhibition rates were 53.5%, 61.4% and 82.6% in PJ34, SAHA and PJ34＋SAHA groups, respectively. The combined use of PJ34 and SAHA could significantly inhibit the xenograft tumor growth when compared with use of P J34 or SAHA alone （P〈0.05）. It was led to conclude that P J34 and SAHA can synergistically suppress the proliferation of liver cancer cells.展开更多
BACKGROUND Colorectal cancer(CRC)is one of the most malignant gastrointestinal cancers worldwide.The liver is the most important metastatic target organ,and liver metastasis is the leading cause of death in patients w...BACKGROUND Colorectal cancer(CRC)is one of the most malignant gastrointestinal cancers worldwide.The liver is the most important metastatic target organ,and liver metastasis is the leading cause of death in patients with CRC.Owing to the lack of sensitive biomarkers and unclear molecular mechanism,the occurrence of liver metastases cannot be predicted and the clinical outcomes are bad for liver metastases.Therefore,it is very important to identify the diagnostic or prognostic markers for liver metastases of CRC.AIM To investigate the highly differentially expressed genes(HDEGs)and prognostic marker for liver metastases of CRC.METHODS Data from three NCBI Gene Expression Omnibus(GEO)datasets were used to show HDEGs between liver metastases of CRC and tumour or normal samples.These significantly HDEGs of the three GEO datasets take the interactions.And these genes were screened through an online tool to explore the prognostic value.Then,TIMER and R package were utilized to investigate the immunity functions of the HDEGs and gene set enrichment analysis was used to explore their potential functions.RESULTS Based on the selection criteria,three CRC datasets for exploration(GSE14297,GSE41258,and GSE49355)were chosen.Venn diagrams were used to show HDEGs common to the six groups and 47 HDEGs were obtained.The HDEGs were shown by using STRING and Cytoscape software.Based on the TCGA database,APOC1 showed significantly different expression between N2 and N0,and N2 and N1.And there was also a significant difference in expression between T2 and T4,and between T2 and T3.In 20 paired CRC and normal tissues,quantitative real-time polymerase chain reaction illustrated that the APOC1 mRNA was strongly upregulated in CRC tissues(P=0.014).PrognoScan and GEPIA2 revealed the prognostic value of APOC1 for overall survival and diseasefree survival in CRC(P<0.05).TIMER showed that APOC1 has a close relationship with immune infiltration(P<0.05).CONCLUSION APOC1 is a biomarker that is associated with both the diagnosis and prognosis of liver metastases of CRC.展开更多
The potential efficacy and clinical feasibility of gene therapy for liver cancer were tested through therecombinant adenovirus-mediated (Ad-multigenes ) co-transfer of human wild-type p53, B7-l co-stimulation(CD8o) an...The potential efficacy and clinical feasibility of gene therapy for liver cancer were tested through therecombinant adenovirus-mediated (Ad-multigenes ) co-transfer of human wild-type p53, B7-l co-stimulation(CD8o) and granulocyte-macrophage colony-stimulating factor (GM-CSF) genes into human hepatocellular carcinoma cell lines. The treated cells underwent apoptosis with specific DNA fragmentation and became more sensitiveto cisplatin, a chemotherapeutic drug. Their growth was partly inhibited. Efficient proliferation and generation ofCTLs and cytokine production were induced in mixed lymphocytes through tumor cell reaction (MLTR) using peripheral blood T lymphocytes from donors as effector cells and Ad-multigenes or Ad-p53-transfected human hepatocellular carcinoma cells (HepG2 or BEL7402) as stimulator cells. Ad-multigenes-transfected rat carcinosarcomaWalker 256 cells were inoculated subcutaneously into normal rats. Fourteen days later, the activity of spleen cellsin rats inoculated with Ad-multigenes-transduced Walker 256 cells was higher than that in Ad-p53-transducedones. These findings suggest that adenovirus-mediated multigenes p53, B7-1 and GM-CSF can induce apoptosis ofliver cancer cells and initiate a potent antitumor immune response against them.展开更多
基金Supported by The 2021 Central-Guided Local Science and Technology Development FundLanzhou COVID-19 Prevention and Control Technology Research Project,No.2020-XG-1Gansu Province Outstanding Graduate Student"Innovation Star"Project,No.2022CXZX-748,No.2022CXZX-746.
文摘BACKGROUND The pyruvate dehydrogenase E1 subunitβ(PDHB)gene which regulates energy metabolism is located in mitochondria.However,few studies have elucidated the role and mechanism of PDHB in different cancers.AIM To comprehensive pan-cancer analysis of PDHB was performed based on bioinformatics approaches to explore its tumor diagnostic and prognostic value and tumor immune relevance in cancer.In vitro experiments were performed to examine the biological regulation of PDHB in liver cancer.METHODS Pan-cancer data related to PDHB were obtained from the Cancer Genome Atlas(TCGA)database.Analysis of the gene expression profiles of PDHB was based on TCGA and Genotype Tissue Expression Dataset databases.Cox regression analysis and Kaplan-Meier methods were used to assess the correlation between PDHB expression and survival prognosis in cancer patients.The correlation between PDHB and receiver operating characteristic diagnostic curve,clinicopathological staging,somatic mutation,tumor mutation burden(TMB),microsatellite instability(MSI),DNA methylation,and drug susceptibility in pan-cancer was also analyzed.Various algorithms were used to analyze the correlation between PDHB and immune cell infiltration and tumor chemotaxis environment,as well as the co-expression analysis of PDHB and immune checkpoint(ICP)genes.The expression and functional phenotype of PDHB in single tumor cells were studied by single-cell sequencing,and the functional enrichment analysis of PDHB-related genes was performed.The study also validated the level of mRNA or protein expression of PDHB in several cancers.Finally,in vitro experiments verified the regulatory effect of PDHB on the proliferation,migration,and invasion of liver cancer.RESULTS PDHB was significantly and differently expressed in most cancers.PDHB was significantly associated with prognosis in patients with a wide range of cancers,including kidney renal clear cell carcinoma,kidney renal papillary cell carcinoma,breast invasive carcinoma,and brain lower grade glioma.In some cancers,PDHB expression was clearly associated with gene mutations,clinicopathological stages,and expression of TMB,MSI,and ICP genes.The expression of PDHB was closely related to the infiltration of multiple immune cells in the immune microenvironment and the regulation of tumor chemotaxis environment.In addition,single-cell sequencing results showed that PDHB correlated with different biological phenotypes of multiple cancer single cells.This study further demonstrated that down-regulation of PDHB expression inhibited the proliferation,migration,and invasion functions of hepatoma cells.CONCLUSION As a member of pan-cancer,PDHB may be a novel cancer marker with potential value in diagnosing cancer,predicting prognosis,and in targeted therapy.
基金This study was supported by the Key Scientific Research Project of Shanghai Municipal Commission of Health and Family Planning(No.201640014)the project of Natural Science Foundation of Jiangxi(No.20171BAB205019)the Special Diseases Program of Pudong New Area Health System(No.PWZzb2017-06).
文摘Objective:Deleted in liver cancer 1(DLC1)is a GTPase-activating protein that is reported as a suppressor in certain human cancers.However,the detailed biological function of DLC1 is still unclear in human prostate cancer(PCa).In the present study,we aimed to explore the function of DLC1 in PCa cells.Methods:Silencing and overexpression of DLC1 were induced in an androgen-sensitive PCa cell line(LNCaP)using RNA interference and lentiviral vector transduction.The Cell Counting Kit-8 assay was performed to determine cell proliferation.The cell cycle was examined by performing a propidium iodide staining assay.Results:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of LNCaP cells.Moreover,DLC1 expression was negatively correlated with Rho-associated protein kinase(ROCK)expression in LNCaP cells.Importantly,this study showed that the ROCK inhibitor Y27632 restored the function of DLC1 in LNCaP cells and reduced the tumorigenicity of LNCaP cells in vivo.Conclusion:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of PCa cells and negatively correlated with ROCK expression in PCa cells and tissue.
文摘目的探讨血清壳多糖酶3样蛋白1(chitinase-3-like protein 1,CHI3L1)、甲胎蛋白(alpha fetoprotein,AFP)和γ-谷氨酰转移酶(γ-glutamyl transferase,GGT)检测在乙型肝炎病毒(hepatitis B virus,HBV)感染相关肝癌诊断中的临床应用价值。方法选取2021年7月—2023年7月在玉林市第一人民医院就诊HBV病毒感染相关的50例肝癌患者,50例肝硬化患者,50例慢性乙型肝炎患者以及50名同期健康体检者作为研究对象。比较4组研究对象血清中CHI3L1、AFP、GGT、丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天门冬氨酸氨基转移酶(aspartate aminotransferase,AST)等水平的差异,用受试者工作特征(receiver operating characteristic curve,ROC)曲线评估各指标在肝癌中的诊断价值。结果肝炎组、肝硬化组、肝癌组的CHI3L1、AST、ALT水平均高于对照组,肝癌组AFP、GGT水平高于对照组,差异有统计学意义(P<0.05)。与肝炎组比较,肝硬化组及肝癌组的CHI3L1、AFP、GGT、AST、ALT水平均升高,差异有统计学意义(P<0.05)。与肝硬化组比较,肝癌组的CHI3L1、AFP、GGT、AST水平均升高,差异有统计学意义(P<0.05)。ROC曲线分析显示,CHI3L1、AFP、GGT联合时的曲线下面积(area under the curve,AUC)最大(AUC=0.936)。Spearman相关分析结果显示,CHI3L1与AST呈正相关(r=0.414,P=0.003),AFP与GGT呈正相关(r=0.437,P=0.002),AFP与AST呈正相关(r=0.504,P<0.001),GGT与AST呈正相关(r=0.759,P<0.001),GGT与ALT呈正相关(r=0.636,P<0.001)。结论CHI3L1、AFP及GGT联合检测可提高肝癌的诊断价值,对临床肝癌患者诊疗有重要作用。
文摘Chronic alcohol consumption is a major risk factor worldwide affecting significantly both mortality and years of life lost （YLL） （1）. Ca. 5% of the western world show risky alcohol consumption and in some countries such as China a regional yearly increase of alcohol consumption of over 400% has been observed recently （2,3）. The liver is the major target organ of alcohol. According to the recently published ＇Global Burden of Disease Study 2010, liver cirrhosis and liver cancer are ranked at position 12 and 16 in the global deaths statistics （1）. Thus, in 2010, ca. 1 million people died from liver cirrhosis with one third directly attributable to alcohol. This is a considerable number when comparing with coronary heart disease with 7 million deaths and the leading cause of mortality. In central Europe, liver cirrhosis even ranks at the fourth position in YLL. Hepatocellular cancer （HCC） is now the most common fatal complication of patients with alcoholic cirrhosis. Moreover, it shows the second fastest increase of all tumors worldwide after kidney tumors and alcohol-associated HCC ranks on third position after HCCs caused by viral hepatitis B and C.
文摘Background:Hepatocellular carcinoma(HCC),the most common type of primary liver cancer,is the fourth leading cause of cancer-related deaths worldwide.Previous evidence shows that the expression of circulating RNA ZFR(circZFR)is upregulated in HCC tissues.However,the molecular mechanism of circZFR in HCC is unclear.Methods:Quantitative reverse transcriptase polymerase chain reaction(qRT-PCR)was employed to detect the expression of circZFR,microRNA-624-3p(miR-624-3p)and WEE1 in HCC tissues and cells.RNase R assay and actinomycin D treatment assay were used to analyze the characteristics of circZFR.For functional analysis,the capacities of colony formation,cell proliferation,cell apoptosis,migration and invasion were assessed by colony formation assay,5-ethynyl-2-deoxyuridine(EdU)assay,flow cytometry assay and transwell assay.Western blot was used to examine the protein levels of WEE1 and epithelial-mesenchymal transition(EMT)-related proteins.The interactions between miR-624-3p and circZFR or WEE1 were validated by dual-luciferase reporter assay and RNA immunoprecipitation(RIP)assay.Xenograft models were established to determine the role of circZFR in vivo.Results:circZFR and WEE1 were upregulated,while miR-624-3p expression was reduced in HCC tissues and cells.circZFR could sponge miR-624-3p,and WEE1 was a downstream gene of miR-624-3p.Knockdown of circZFR significantly reduced the malignant behaviors of HCC and that co-transfection with miR624-3p inhibitor restored this change.Overexpression of WEE1 abolished the inhibitory effect of miR624-3p mimic on HCC cells.Mechanistically,circZFR acted as a competitive endogenous RNA(ceRNA)to regulate WEE1 expression by targeting miR-624-3p.Furthermore,in vivo studies have illustrated that circZFR knockdown inhibited tumor growth.Conclusions:circZFR knockdown reduced HCC cell proliferation,migration and invasion and promoted apoptosis by regulating the miR-624-3p/WEE1 axis,suggesting that the circZFR/miR-624-3p/WEE1 axis might be a potential target for HCC treatment.
基金Supported by National Natural Science Foundation of China,No.81472213the Health Commission of Zhejiang Province,No.2019ZD010 and No.2019ZD029+1 种基金the Science Technology Department of Zhejiang Province,No.LGF20H220001the Zhejiang Provincial Administration of Traditional Chinese Medicine,No.2021ZA088.
文摘BACKGROUND Activated hepatic stellate cells(aHSCs)are the major source of cancer-associated fibroblasts in the liver.Although the crosstalk between aHSCs and colorectal cancer(CRC)cells supports liver metastasis(LM),the mechanisms are largely unknown.AIM To explore the role of BMI-1,a polycomb group protein family member,which is highly expressed in LM,and the interaction between aHSCs and CRC cells in promoting CRC liver metastasis(CRLM).METHODS Immunohistochemistry was carried out to examine BMI-1 expression in LM and matched liver specimens of CRC.The expression levels of BMI-1 in mouse liver during CRLM(0,7,14,21,and 28 d)were detected by Western blotting(WB)and the quantitative polymerase chain reaction(qPCR)assay.We overexpressed BMI-1 in HSCs(LX2)by lentivirus infection and tested the molecular markers of aHSCs by WB,qPCR,and the immunofluorescence assay.CRC cells(HCT116 and DLD1)were cultured in HSC-conditioned medium(LX2 NC CM or LX2 BMI-1 CM).CM-induced CRC cell proliferation,migration,epithelial-mesenchymal transition(EMT)phenotype,and transforming growth factor beta(TGF-β)/SMAD pathway changes were investigated in vitro.A mouse subcutaneous xenotransplantation tumor model was established by co-implantation of HSCs(LX2 NC or LX2 BMI-1)and CRC cells to investigate the effects of HSCs on tumor growth and the EMT phenotype in vivo.RESULTS Positive of BMI-1 expression in the liver of CRLM patients was 77.8%.The expression level of BMI-1 continued to increase during CRLM in mouse liver cells.LX2 overexpressed BMI-1 was activated,accompanied by increased expression level of alpha smooth muscle actin,fibronectin,TGF-β1,matrix metalloproteinases,and interleukin 6.CRC cells cultured in BMI-1 CM exhibited enhanced proliferation and migration ability,EMT phenotype and activation of the TGF-β/SMAD pathway.In addition,the TGF-βR inhibitor SB-505124 diminished the effect of BMI-1 CM on SMAD2/3 phosphorylation in CRC cells.Furthermore,BMI-1 overexpressed LX2 HSCs promoted tumor growth and the EMT phenotype in vivo.CONCLUSION High expression of BMI-1 in liver cells is associated with CRLM progression.BMI-1 activates HSCs to secrete factors to form a prometastatic environment in the liver,and aHSCs promote proliferation,migration,and the EMT in CRC cells partially through the TGF-β/SMAD pathway.
基金Supported by State Key Project for Infectious Diseases,No.2013ZX10002009 and No.2012ZX10002-017State Key Project for Basic Research,No.2014CBA02001 and No.2014CBA02002+1 种基金National High-tech R and D Program,No.2012AA020206Natural Science Foundation of China,No.81071789 and No.81321091
文摘AIM: To investigate the expression characteristics of peroxiredoxin 1(PRDX1) m RNA and protein in liver cancer cell lines and tissues.METHODS: The RNA sequencing data from 374 patients with liver cancer were obtained from The Cancer Genome Atlas. The expression and clinical characteristics of PRDX1 m RNA were analyzed in this dataset. The Kaplan-Meier and Cox regression survival analysis was performed to determine the relationship between PRDX1 levels and patient survival. Subcellular fractionation and Western blotting were used to demonstrate the expression of PRDX1 protein in six liver cancer cell lines and 29 paired fresh tissue specimens. After bioinformatics prediction,a putative posttranslational modification form of PRDX1 was observed using immunofluorescence under confocal microscopy and immunoprecipitation analysis in liver cancer cells.RESULTS: The m RNA of PRDX1 gene was upregulated about 1.3-fold in tumor tissue compared with the adjacent non-tumor control(P = 0.005). Its abundance was significantly higher in men than women(P < 0.001). High levels of PRDX1 m RNA were associated with a shorter overall survival time(P =0.04) but not with recurrence-free survival. The Cox regression analysis demonstrated that patients with high PRDX1 m RNA showed about 1.9-fold increase of risk for death(P = 0.03). In liver cancer cells,PRDX1 protein was strongly expressed with multiple different bands. PRDX1 in the cytosol fraction existed near the theoretical molecular weight,whereas two higher molecular weight bands were present in the membrane/organelle and nuclear fractions. Importantly,the theoretical PRDX1 band was increased,whereas the high molecular weight form was decreased in tumor tissues. Subsequent experiments revealed that the high molecular weight bands of PRDX1 might result from the post-translational modification by small ubiquitin-like modifier-1(SUMO1).CONCLUSION: PRDX1 was overexpressed in the tumor tissues of liver cancer and served as an independent poor prognostic factor for overall survival. PRDX1 can be modified by SUMO to play specific roles in hepatocarcinogenesis.
基金supported by grants from the National Natural Science Foundation of China (81041025 and 81000189)
文摘BACKGROUND:Phospholipid scramblase 1(PLSCR1) not only participates in the transbilayer movement of phospholipids,but also plays a role in the pathogenesis and progression of cancers.The present study aimed to evaluate the effect of silencing PLSCR1 expression by RNA interference in colorectal cancer(CRC) and metastatic liver cancer.METHODS:The expression of PLSCR1 in CRC and metastatic liver cancer samples was assessed by immunohistochemistry.The cultured cells with the highest expression were selected for subsequent experiments.We designed three siRNA oligonucleotide segments targeted at PLSCR1.Successful transfection was confirmed.The biological behavior of the cells in proliferation,adhesion,migration and invasion was determined.RESULTS:PLSCR1 protein expression increased significantly in the majority of CRC and metastatic liver cancer samples compared with normal samples.Lovo cells had the highest expression of PLSCR1.The siRNA-390 oligonucleotide segment had the best silencing effect.After transfection,Lovo cell proliferation was significantly inhibited compared with the controls in the MTT assay.Laminin and fibronectin adhesion assays showed Lovo cell adhesion was also significantly inhibited.In the migration assay,the number of migrating cells in the PLSCR1 siRNA-390 group was 50±12,significantly lower than the number in the siRNA-N group(115±28) and in the control group(118±31).In an invasion test,the number of invading cells in the PLSCR1 siRNA-390 group was 60±18,significantly lower than that in the siRNA-N group(97±26) and the control group(103±24).CONCLUSIONS:PLSCR1 is overexpressed in CRC and metastatic liver cancer.Silencing of PLSCR1 by siRNA inhibits the proliferation,adhesion,migration and invasion of Lovo cells,which suggests that PLSCR1 contributes to the tumorigenesis and tumor progression of CRC.PLSCR1 may be a potential gene therapy target for CRC and associated metastatic liver cancer.
基金supported by grants from the National Natural Science Foundation of China(No.81172293)the New Century Excellent Talent Foundation of Ministry of Education of China(No.NCET-04-0701)
文摘Summary： Poly （ADP-ribose） polymerase-1 （PARP-1） inhibitors and histone deacetylase （HDAC） in- hibitors have recently emerged as promising anticancer drugs. The aim of this study was to investigate the effect of combination treatment with the PARP inhibitor P J34 and HDAC inhibitor SAHA on the proliferation of liver cancer cells. Cell proliferation and apoptosis were assessed in three human liver cancer cell lines （HepG2, Hep3B and HCC-LM3） treated with PJ34 （8 μmol/L） and SAHA （1 panol/L）, alone or combined, by Cell Counting Kit-8 assay and flow cytometry, respectively. The nude mice bear- ing subcutaneous HepG2 tumors were administered different groups of drugs （10 mg/kg PJ34, 25 mg/kg SAHA, 10 mg/kg PJ34＋25 mg/kg SAHA）, and the inhibition rates of tumor growth were compared between groups. The results showed that combined use of P J34 and SAHA could synergistically inhibit the proliferation of liver cancer cell lines HepG2, Hep3B and HCC-LM3. The apoptosis rate of HepG2 cells treated with PJ34＋SAHA was significantly higher than that of HepG2 cells treated with PJ34 or SAHA alone （P〈0.05）. In vivo, the tumor inhibition rates were 53.5%, 61.4% and 82.6% in PJ34, SAHA and PJ34＋SAHA groups, respectively. The combined use of PJ34 and SAHA could significantly inhibit the xenograft tumor growth when compared with use of P J34 or SAHA alone （P〈0.05）. It was led to conclude that P J34 and SAHA can synergistically suppress the proliferation of liver cancer cells.
基金National Key Research and Development(R&D)Program Project,No.2019YFC1315705.
文摘BACKGROUND Colorectal cancer(CRC)is one of the most malignant gastrointestinal cancers worldwide.The liver is the most important metastatic target organ,and liver metastasis is the leading cause of death in patients with CRC.Owing to the lack of sensitive biomarkers and unclear molecular mechanism,the occurrence of liver metastases cannot be predicted and the clinical outcomes are bad for liver metastases.Therefore,it is very important to identify the diagnostic or prognostic markers for liver metastases of CRC.AIM To investigate the highly differentially expressed genes(HDEGs)and prognostic marker for liver metastases of CRC.METHODS Data from three NCBI Gene Expression Omnibus(GEO)datasets were used to show HDEGs between liver metastases of CRC and tumour or normal samples.These significantly HDEGs of the three GEO datasets take the interactions.And these genes were screened through an online tool to explore the prognostic value.Then,TIMER and R package were utilized to investigate the immunity functions of the HDEGs and gene set enrichment analysis was used to explore their potential functions.RESULTS Based on the selection criteria,three CRC datasets for exploration(GSE14297,GSE41258,and GSE49355)were chosen.Venn diagrams were used to show HDEGs common to the six groups and 47 HDEGs were obtained.The HDEGs were shown by using STRING and Cytoscape software.Based on the TCGA database,APOC1 showed significantly different expression between N2 and N0,and N2 and N1.And there was also a significant difference in expression between T2 and T4,and between T2 and T3.In 20 paired CRC and normal tissues,quantitative real-time polymerase chain reaction illustrated that the APOC1 mRNA was strongly upregulated in CRC tissues(P=0.014).PrognoScan and GEPIA2 revealed the prognostic value of APOC1 for overall survival and diseasefree survival in CRC(P<0.05).TIMER showed that APOC1 has a close relationship with immune infiltration(P<0.05).CONCLUSION APOC1 is a biomarker that is associated with both the diagnosis and prognosis of liver metastases of CRC.
文摘The potential efficacy and clinical feasibility of gene therapy for liver cancer were tested through therecombinant adenovirus-mediated (Ad-multigenes ) co-transfer of human wild-type p53, B7-l co-stimulation(CD8o) and granulocyte-macrophage colony-stimulating factor (GM-CSF) genes into human hepatocellular carcinoma cell lines. The treated cells underwent apoptosis with specific DNA fragmentation and became more sensitiveto cisplatin, a chemotherapeutic drug. Their growth was partly inhibited. Efficient proliferation and generation ofCTLs and cytokine production were induced in mixed lymphocytes through tumor cell reaction (MLTR) using peripheral blood T lymphocytes from donors as effector cells and Ad-multigenes or Ad-p53-transfected human hepatocellular carcinoma cells (HepG2 or BEL7402) as stimulator cells. Ad-multigenes-transfected rat carcinosarcomaWalker 256 cells were inoculated subcutaneously into normal rats. Fourteen days later, the activity of spleen cellsin rats inoculated with Ad-multigenes-transduced Walker 256 cells was higher than that in Ad-p53-transducedones. These findings suggest that adenovirus-mediated multigenes p53, B7-1 and GM-CSF can induce apoptosis ofliver cancer cells and initiate a potent antitumor immune response against them.
