Effects of 9-cis retinoic acid on human homeobox gene NKX3.1 expression in prostate cancer cell line LNCaP

Aim:To study the regulatory effects of 9-cis retinoic acid(RA)on the expression of human homeobox gene NKX3.1 in prostate cancer cell line LNCaP.Methods:Flow cytometry,reverse transcriptase polymerase chain reaction and Western blotting were performed to evaluate the effects of 9-cis RA on NKX3.1 expression and cell cycle of LNCaP cells.To identify a regulatory region within the NKX3.1 promoter contributing to the regulation induced by 9-cis RA, we have constructed an NKX3.1 promoter-reporter plasmid,pGL_3-1040bp,and its 5′-deletion mutants,which were transfected into LNCaP cells with treatment of 9-cis RA in indicated concentrations.Results:With the treatment of 9-cis RA,the NKX3.1 promoter activity was increased in reporter gene assay and NKX3.1 expression was enhanced at both mRNA and protein levels in LNCaP cells.We found that the region between -936 and -921 in the upstream of NKX3.1 gene involved the inducible regulation by 9-cis RA treatment.In flow cytometry,9-cis RA treatment caused accumulation of cells in the G_1 phase of the cell cycle and a fewer cells pass through to G_2/M.Conclusion:Our results demonstrated that 9-cis RA as a differentiating agent can arrest prostate cancer cells in G_1 phase and reduce cell mitosis,and upregulate the expression of human homeobox gene NKX3.1,which is thought to play an important role in prostate differentiation and to act as a tumor suppressor gene in the prostate.(Asian J Androl 2006 Jul;8:435-441)

Association between endogenous gene expression and growth regulation induced by TGF-β1 in human gastric cancer cells

AIM: To investigate the association between endogenous gene expression and growth regulation including proliferation and apoptosis induced by transforming growth factor-pi (TGF-βl) in human gastric cancer (GC) cells. METHODS: Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the main components of the TGF-β1/Smads signal pathway in human poorly differentiated GC cell line BGC-823. Localization of Smad proteins was also determined using immunofluorescence. Then, the BGC-823 cells were cultured in the presence or absence of TGF-β1 (10 ng/mL) for 24 and 48 h, and the effects of TGF-β1 on proliferation and apoptosis were measured by cell growth curve and flow cytometry (FCM) analysis. The ultrastructural features of BGC-823 cells with or without TGF-β1 treatment were observed under transmission electron microscope. The apoptotic cells were visualized by means of the terminal deoxynucleotidyl transferase (TdT)-mediated dTUP in situ nick end-labeling (TUNEL) method. Meanwhile, the expression levels of endogenous p15, p21 and Smad7 mRNA and the corresponding proteins in the cells were detected at 1, 2 and 3 h after culture in the presence or absence of TGF-β1 (10 ng/mL) by semi-quantitative RT-PCR and Western blot, respectively. RESULTS: The TGF-β1/Smad signaling was found to be intact and functional in BGC-823 cells. The growth curve revealed the most evident inhibition of cell proliferation by TGF-β1 at 48 h, and FCM assay showed G1 arrest accompanied with apoptosis induced by TGF-β1. The typical morphological changes of apoptosis were observed in cells exposed to TGF-β1. The apoptosis index (AI) in TGF-β1-treated cells was significantly higher than that in the untreated controls (10.7±1.3% vs 0.32±0.06%, P＜0.01). The levels of p15, p21 and Smad7mRNA and corresponding proteins in cells were significantly up-regulated at 1 h, but gradually returned to basal levels at 3 h following TGF-βl (10 ng/mL) treatment. CONCLUSION: TGF-β1 affects both proliferation and apoptosis of GC cells through the regulation of p15 and p21, and induces transient expression of Smad 7 as a negative feedback modulation of TGF-β1 signal. Our results suggest a novel functional role of p21 as an accelerant of TGF-β1-mediated apoptosis in GC cells.

Association of hypoxia-inducible factor-1α (HIF1α) 1772C/T genepolymorphism with susceptibility to renal cell carcinoma/prostatecancer

In this study,we used a meta-analysis method to evaluate the relationship between hypoxia-inducible factor-1α(HIF1α)1772C/T gene polymorphism(rs 11549465)and renal cell carcinoma(RCC)/prostate cancer risk.We searched for relevant studies(before March 1,2019)on Cochrane Library,Embase,and PubMed.Studies meeting the inclusion criteria were recruited into this meta-analysis.The outcome of dichotomous data was showed in the way of odds ratios(OR),and 95%confidence intervals(CI)were also counted.In this investigation,there was no association between HIF1α1772C/T gene polymorphism and susceptibility to RCC in Caucasians,Asians as well as overall populations.In addition,HIF1α1772C/T gene polymorphism was not found to be relevant to the survival in RCC.Interestingly,the T allele was relevant to prostate cancer risk in all populations,but not in Caucasians and Asians.However,the TT genotype and the CC genotype were not related to prostate cancer susceptibility in Asian,Caucasian,and all populations.In conclusion,the T allele of the HIF1α1772C/T gene polymorphism was related to prostate cancer risk in the overall populations.

Prostate cancer antigen-1 as a cancer potential novel marker for prostate

Aim： To examine the expression of prostate cancer antigen-1 （PCA-1） in prostate cancer （PCa） and to validate it as a potential marker for diagnosis of PCa. Methods： In situ hybridization analysis of PCA-1 mRNA expression was performed on 40 benign prostate hyperplasia （BPH）, 16 high-grade prostatic intraepithelial neoplasm （HG-PIN）, 74 PCa and 34 other malignant carcinoma specimens. The level of PCA- 1 expression was semiquanfitatively scored by assessing both the percentage and intensity of PCA- 1 positive staining cells in the specimens. We then compared the PCA-1 expression between BPH, HG-PIN and PCa and evaluated the correlation of PCA-1 expression level with clinical parameters of PCa. Results： PCA-1 mRNA was expressed in the majority of both PCa and HG-PIN specimens but not in BPH and other malignant carcinoma. The expression level of PCA-1 increased along with a high Gleason score （P 〈 0.05）, and was unrelated to other clinical parameters of PCa （all P 〉 0.05）. Conclusion： The data suggest that PCA-1 might be a novel diagnostic marker for PCa, and that increased PCA-1 expression might denote more aggressive variants of PCa.

The expressional level of tankyrase-1 gene and its regulation in colorectal cancer in a Saudi population

Tankyrase1(TNKS1)plays an essential role in cancer progression by regulating telomere length.The study aimed to determine expression of TNKS1 and its regulation in colorectal cancer(CRC)in 20 samples from Saudi patients.mRNA expression of TNKS1 in CRC and paired normal tissues was measured by qRT-PCR.Epigenetic modification of TNKS1 promoter was determined by methylation-specific PCR while somatic mutation was analyzed by Sanger sequencing in exon 10 of the gene.All cancerous and normal tissues expressed TNKS1,but level of expression in CRC tissues was significantly associated with tumor stage though no other parameters;age,gender,and tumor location,showed any correlation.Expression of TNKS1 was markedly higher in earlier(I,II)than in later(Ⅲ,Ⅳ)stages of CRC development.Both cancerous and healthy tissues had unmethylated promoters.Sanger sequencing of exon 10 masked any somatic mutation in the samples.Our findings suggest that up-regulation of TNKS1 was inversely correlated with cancer progression in CRC,indicating that TNKS1 participates in the initiation of CRC by stabilizing telomere length in the first phase of cancer progression.Mechanisms other than TNKS1 might play a role in malignant tumor progression and telomere maintenance in the late stages of CRC.

Gene Expression Profile, Androgen Independence and Prostate Cancer

It is now generally accepted that the burden of disease due to prostate cancer has tremendously increased globally. Current data indicates that prostate cancer is the most common form of cancer in men in the United State of America, and the second leading cause of death due to cancer in men. Progression to androgen independence and subsequent therapeutic resistance and death is a common fate of patients with prostate cancer. This review highlights the gene expression profile of androgen independent prostate cancer and the possible mechanisms that results in transformation to such treatment resistant state.

Correlation of pituitary tumor transforming gene 1 with proliferation and invasion genes in prostate cancer

Objective:To study the change of pituitary tumor transforming gene 1 (PTTG1) expression in prostate cancer and its correlation with proliferation and invasion genes.Methods: Patients with prostate cancer who underwent radical operation in our hospital between March 2015 and January 2018 were selected as the malignant group of the research, and the prostate cancer lesions were collected;patients who underwent transurethral resection of the prostate due to benign prostatic hyperplasia in our hospital during the same period were selected as the benign group of the research, and the benign prostate lesions were collected. The mRNA expression levels of PTTG1, proliferation genes and invasion genes in the lesions were determined. Results:PTTG1, Survivin, Bcl-2, CyclinD1, GPRC6A, ZEB1, CatB, CatD and PAR-1 mRNA expression in prostate cancer lesions of malignant group were significantly higher than those of benign group whereas CDKN2, p21 and TFPI2 mRNA expression were significantly lower than those of benign group;Survivin, Bcl-2, CyclinD1, GPRC6A, ZEB1, CatB, CatD and PAR-1 mRNA expression in prostate cancer lesions with high PTTG1 were significantly higher than those in prostate cancer lesions with low PTTG1 whereas CDKN2, p21 and TFPI2 mRNA expression were significantly lower than those in prostate cancer lesions with low PTTG1.Conclusion:The PTTG1 gene is highly expressed in prostate cancer lesions and it is closely related to the changes of proliferation and invasion gene expression.

LncRNA MEG3 acts a biomarker and regulates cell functions by targeting ADAR1 in colorectal cancer

BACKGROUND Colorectal cancer (CRC) is the third most prevalent malignancy and has the fourth highest global cancer mortality rate. Early diagnosis and prompt medical attention can improve quality of life and the prognosis of CRC patients. Accumulating evidence reveals that long non-coding RNAs (lncRNAs) function as oncogenes or anti-oncogenes, as well as biomarkers in various cancers. AIM To investigate the levels and molecular mechanism of the lncRNA maternally expressed gene 3 (MEG3) in CRC. METHODS The levels of lncRNA MEG3 in CRC tissue, serum and cell line samples were explored via qRT-PCR. The relationship between MEG3 levels and clinicopathological features in CRC was investigated. The diagnostic and prognostic values of serum MEG3 levels were analyzed with ROC curves and KaplanMeier survival curves, respectively. RESULTS Significant decreased levels of MEG3 existed in CRC tissue, cell lines and serum. CRC patients with down-regulated serum MEG3 levels had larger tumor sizes, and advanced clinical stages. The sensitivity and specificity of serum MEG3 levels in CRC detection was 0.667 and 0.875, respectively. Tumor size, T stages, and serum MEG3 levels are indie factors that produce an effect on CRC patients' prognosis. KaplanMeier survival curves suggested that CRC patients with high levels of MEG3 had a remarkably better overall survival rate. CONCLUSION LncRNA MEG3 is down-regulated in CRC, and regulates cell functions by targeting adenosine deaminase’s effect on RNA 1 in CRC.

Differences in phenotype and gene expression of prostate stromal cells from patients of varying ages and their influence on tumour formation by prostate epithelial cells

Prostate cancer （PCa） is an age-related disease, and the stromal microenvironment plays an important role in prostatic malignant progression. However, the differences in prostate stromal cells present in young and old tissue are still obscure. We established primary cultured stromal cells from normal prostatic peripheral zone （PZ） of donors of varying ages and found that cultured stromal cells from old donors （PZ-old） were more enlarged and polygonal than those from young donors （PZ-young）. Furthermore, based on immunocytochemical and ultrastructural analysis, the components of stromal cells changed from a majority of fibroblasts to a mixture of fibroblasts and myofibroblasts with increasing donor age. Using a three-dimensional in vitro culture system, we found that PZ-old stromal cells could enhance the proliferation, migration and invasion of cocultured benign BPH-1 and PC-3 cells. Using an in vivo tissue recombination system, we also found that PZ-old stromal cells are more effective than PZ-young cells in promoting tumour formation by BPH-1 cells of high passage（〉100） and PC-3 cells. To probe the possible mechanism of these effects, we performed cDNA microarray analysis and profiled 509 upregulated genes and 188 downregulated genes in PZ-old cells. Among the changed genes, we found genes coding for a subset of paracrine factors that are capable of influencing adjacent epithelial cells; these include hepatocyte growth factor （HGF）, fibroblast growth factor 5 （FGF5）, insulin-like growth factor 2 （IGF2）, insulin-like growth factor-binding protein 4 （IGFBP4）, IGFBP5 and matrix metallopeptidase 1 （MMP1）. Changes in the expression of these genes were further confirmed by quantitative real-time polymerase chain reaction （PCR）, Western blotting and enzyme-linked immunosorbent assays. Overall, our findings indicate that stromal cells from prostate PZ of old donors are more active than similar cells from young donors in promoting the malignant process of adjacent epithelial cells. This finding hints at a new potential strategy for the prevention of PCa.

Discovery and validation of prognostic markers in gastric cancer by genome-wide expression profiling

AIM: To develop a prognostic gene set that can predict patient overall survival status based on the whole genome expression analysis. METHODS: Using Illumina HumanWG-6 BeadChip followed by semi-supervised analysis, we analyzed the expression of 47 296 transcripts in two batches of gastric cancer patients who underwent surgical resection. Thirty-nine samples in the first batch were used as the training set to discover candidate markers correlated to overall survival, and thirty-three samples in the second batch were used for validation. RESULTS: A panel of ten genes were identified as prognostic marker in the first batch samples and classified patients into a lowand a high-risk group with significantly different survival times (P = 0.000047). This prognostic marker was then verified in an independent validation sample batch (P = 0.0009). By comparing with the traditional Tumor-node-metastasis (TNM) staging system, this ten-gene prognostic marker showed consistent prognosis results. It was the only independent prognostic value by multivariate Cox regression analysis (P = 0.007). Interestingly, six of these ten genes are ribosomal proteins, suggesting a possible association between the deregulation of ribosome related gene expression and the poor prognosis. CONCLUSION: A ten-gene marker correlated with overall prognosis, including 6 ribosomal proteins, was identified and verified, which may complement the predictive value of TNM staging system.

ODC1基因对前列腺癌细胞PC-3 LncRNA表达影响的初步观察与分析

目的探讨鸟氨酸脱羧酶1(ODC1)基因对前列腺癌细胞PC-3 LncRNA表达的影响,为进一步研究其作用及调控机制提供一些实验基础和线索。方法以前列腺癌细胞PC-3作为研究对象,敲低ODC1基因,经q-PCR检测及WB检测验证后采用Illumina Novaseq 6000,PE150模式进行高通量测序,测序结果应用编码潜能分析方法(CPC分析、CNCI分析、CPAT分析、LGC分析)进行预测筛选,对筛选出的lncRNAs行差异分析、顺式作用元件分析以及GO和KEGG富集分析。再选择文献报道中与前列腺癌关系密切的几个功能lncRNAs,采用q-PCR进行验证。结果(1)在ODC1敲低PC-3细胞中筛选到341个lncRNA,差异表达的lncRNA中上调154个,下调128个。符合条件的差异lncRNA与差异表达基因共表达的顺式调控靶标有38个。(2)差异表达lncRNAs行GO和KEGG富集分析显示,lncRNA靶标基因参与体内多种生物学通路,如磷脂酰肌醇3激酶/蛋白激酶B信号通路(PI3K-Akt信号通路)、Ras信号通路等,还与肿瘤血管生成、细胞群增殖、正调控细胞分裂等密切相关。(3)q-PCR法验证结果:LINC00973、TERC表达显著下调、LINC00638表达显著上调,与GO和KEGG富集分析得到的lncRNA差异表达情况一致。结论敲低ODC1基因可导致PC-3细胞lncRNAs差异表达,ODC1基因有可能通过lncRNAs影响细胞信号通路,促进肿瘤的发生发展。本实验为进一步研究ODC1基因在前列腺癌中的作用提供了一些新的实验基础。

Differential analysis revealing APOC1 to be a diagnostic and prognostic marker for liver metastases of colorectal cancer

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.

谷胱甘肽S转移酶P1和X线修复交错互补基因1基因多态性与前列腺癌化疗敏感性及预后关系研究

目的:探究前列腺癌患者谷胱甘肽S转移酶P1(GSTP1)和X线修复交错互补基因1(XRCC1)基因多态性与化疗敏感性及预后的关系。方法:选取2018年5月至2021年5月行雄激素剥夺治疗+双药化疗的前列腺癌患者103例,收集患者临床资料,采用PCR-PFLP方法对患者行基因分型,并分析其GSTP1-rs1695位点和XRCC1-rs25487位点的多态性,分析其与化疗敏感性的关系,并探究GSTP1和XRCC1基因多态性与患者3年生存率的相关性。结果:103例前列腺癌化疗患者GSTP1-rs1695位点和XRCC1-rs25487位点分布均符合Hardy-Weinberg平衡(χ2=9.794,P>0.05)。GSTP1-rs1695位点中AA基因型占比为65.05%(67/103),AG基因型占比为23.30%(24/103),GG基因型占比为11.65%(12/103);XRCC1-rs25487位点中AA基因型占比为29.13%(30/103),AG基因型占比为50.49%(52/103),GG基因型占比为20.39%(21/103)。GSTP1-rs1695 AA型化疗敏感性为35.82%,低于AG/GG型58.33%(P<0.05)。XRCC1-rs25487 AA型化疗敏感性为40.00%,AG/GG型45.21%,两者差异无统计学意义(P>0.05)。GSTP1-rs1695、XRCC1-rs25487不同表型患者3年无进展生存率之间无明显差异(P>0.05)。GSTP1-rs1695 AA型3年总生存率低于AG/GG型;XRCC1-rs25487 AA型低于AG/GG型(P<0.05)。多因素COX回归分析结果显示,GSTP1-rs1695 AA型、XRCC1-rs25487 AA型均为前列腺癌患者化疗后3年总生存期的独立影响因素。结论:GSTP1-rs1695、XRCC1-rs25487基因多态性对前列腺癌患者化疗敏感性和预后均有一定影响,GSTP1-rs1695和XRCC1-rs25487 A等位基因突变均可导致更短的3年生存率,G等位基因突变可带来更好的化疗敏感性和生存期。

NDRG1过表达对去势抵抗性前列腺癌耐药细胞株C4-2/ENZA耐药性的影响及其机制

目的:探讨N-myc下游调节基因1(NDRG1)对去势抵抗性前列腺癌(CRPC)恩杂鲁胺(ENZA)耐药的影响,并阐明其作用机制。方法:体外培养人CRPC C4-2细胞和ENZA耐药株C4-2/ENZA细胞,采用实时荧光定量PCR(RT-qPCR)法检测C4-2/ENZA细胞及其亲本C4-2细胞中NDRG1 mRNA表达水平,Western blotting法检测C4-2/ENZA细胞及其亲本C4-2细胞中NDRG1、雄激素受体(AR)和前列腺特异性抗原(PSA)蛋白表达水平,以验证细胞转染效率。将C4-2/ENZA细胞分为空白组(正常培养不进行处理)、阴性对照慢病毒(Lv-NC)组(转染Lv-NC)、Lv-NDRG1组(转染Lv-NDRG1)、Lv-NC+ENZA组(转染Lv-NC后加入ENZA处理)、Lv-NDRG1+ENZA组(转染Lv-NDRG1后加入ENZA处理)、Lv-NDRG1+表皮生长因子(EGF)组(转染Lv-NDRG1后加入EGF处理)和Lv-NDRG1+EGF+ENZA组(转染Lv-NDRG1后加入EGF和ENZA处理)。采用噻唑蓝(MTT)法检测各组细胞半数抑制浓度(IC_(50))、耐药指数(RI)和细胞增殖活性,流式细胞术检测各组细胞凋亡率,RT-qPCR法检测各组细胞中NDRG1 mRNA表达水平,Western blotting法检测各组细胞中NDRG1、AR、第213位丝氨酸磷酸化雄激素受体(p-ARSer^(213))、第81位丝氨酸磷酸化雄激素受体(p-ARSer^(81))和PSA蛋白表达水平。结果:与C4-2细胞比较,C4-2/ENZA细胞中NDRG1 mRNA和蛋白表达水平均明显降低(P<0.01),AR和PSA蛋白表达水平明显升高(P<0.01),提示ENZA耐药株C4-2/ENZA中NDRG1低表达;与Lv-NC组比较,LvNDRG1组细胞中NDRG1 mRNA和蛋白表达水平均明显升高(P<0.01),提示成功构建NDRG1基因过表达C4-2/ENZA耐药细胞株。MTT法,与C4-2细胞比较,C4-2/ENZA细胞IC_(50)明显升高(P<0.01),RI为17.78;与Lv-NC组比较,Lv-NDRG1组C4-2/ENZA细胞IC_(50)明显降低(P<0.01)。EGF处理24 h,与Lv-NC组比较,Lv-NC+EGF组C4-2/ENZA细胞IC_(50)明显升高(P<0.01);与Lv-NDRG1组比较,Lv-NDRG1+EGF组C4-2/ENZA细胞IC_(50)明显升高(P<0.01)。与ENZA处理前比较,不同浓度ENZA处理24 h,C4-2和C4-2/ENZA细胞增殖活性均逐渐降低(F=223.80,P<0.01;F=81.46,P<0.01)。ENZA处理24 h,与Lv-NC组比较,Lv-NDRG1组C4-2/ENZA细胞增殖活性明显降低(P<0.01)。EGF处理24 h,与Lv-NC组比较,Lv-NC+EGF组C4-2/ENZA细胞增殖活性明显升高(P<0.01),Lv-NDRG1+EGF组C4-2/ENZA细胞增殖活性明显降低(P<0.01)。选择10.000μmol·L^(-1) ENZA和干预24 h作为后续检测浓度及时间点。流式细胞术,ENZA处理24 h,与Lv-NC组比较,Lv-NDRG1组细胞凋亡率明显升高(P<0.01);与Lv-NC+ENZA组比较,Lv-NDRG1+ENZA组细胞凋亡率明显升高(P<0.01)。EGF处理24h,与Lv-NDRG1组比较,Lv-NDRG1+EGF组细胞凋亡率明显降低(P<0.01),Lv-NDRG1+ENZA组细胞凋亡率明显升高(P<0.01);与Lv-NDRG1+ENZA组比较,Lv-NDRG1+EGF+ENZA组细胞凋亡率明显降低(P<0.01)。Western blotting法,ENZA处理24 h,与Lv-NC组比较,Lv-NDRG1组细胞中AR和PSA蛋白表达水平及p-ARSer^(213)/AR和p-ARSer^(81)/AR比值均明显降低(P<0.05或P<0.01)。EGF处理24 h,与Lv-NC组比较,Lv-NC+EGF组细胞中AR和PSA蛋白表达水平及p-ARSer^(213)/AR和p-ARSer^(81)/AR比值均明显升高(P<0.05或P<0.01);与Lv-NDRG1组比较,Lv-NDRG1+EGF组细胞中AR和PSA蛋白表达水平及p-ARSer^(213)/AR和p-ARSer^(81)/AR比值均明显升高(P<0.01)。结论:NDRG1过表达可降低CRPC对ENZA的耐药性,其作用机制可能与抑制AR信号转导有关。

Differential gene screening and functional analysis in docetaxel-resistant prostate cancer cell lines

Objective Docetaxel-based combination chemotherapy has traditionally been the standard treatment for metastatic castration-resistant prostate cancer(PCa).However,most patients eventually develop resistance to this treatment,which further reduces their survival.This study aimed to determine key molecular genes in docetaxel-resistant PCa cell lines using bioinformatic approaches.Methods The analysis of microarray data GSE33455(including DU-145/DU-145R and PC-3/PC-3R cell lines)obtained from the Gene Expression Omnibus(GEO)database was performed using GEO2R.Differentially expressed genes(DEGs)of DU-145/DU-145R and PC-3/PC-3R cell lines were selected,and the intersection of DEGs between the two groups was obtained.DEGs were annotated with the Gene Ontology(GO)function and enriched with the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway using an online platform(https://cloud.oebiotech.cn/task/detail/array_enrichment/).The online tool Search Tool for the Retrieval of Interacting Genes(https://string-db.org/)was used to obtain the DEG network graph and matrix list,which was imported into Cytoscape 3.6.1 and analyzed using the Molecular Complex Detection plug-in to detect potential functional modules in the network.Results A total of 131 intersection DEGs were identified between non-treated and docetaxel-resistant PCa cell lines.GO functional annotation showed that the main genes involved were present in the plasma membrane and were involved in positive regulation of ubiquitin-protein transferase activity,positive regulation of pseudopodium assembly,centriolar subdistal appendage,and heterophilic cell-cell adhesion via plasma membrane cell adhesion molecules.KEGG pathway enrichment analysis revealed that DEGs were mainly involved in IL-17 signaling pathway,cytokine-cytokine receptor interaction,rheumatoid arthritis,legionellosis,and folate biosynthesis.We identified two distinct hubs of DEGs:(1)CD274,C-X-C motif chemokine ligand(CXCL)1,DExD/H-box helicase 58,CXCL2,CXCL8,colony-stimulating factor 2,C-X-C motif chemokine receptor 4(CXCR4),CXCL5,and CXCL6 and(2)argininosuccinate lyase,argininosuccinate synthase 1,and asparagine synthetase.Except for the CXCR4 gene that was downregulated,the other 11 genes showed upregulated expression.Conclusion Certain differential genes may be potential targets for predicting and treating metastatic docetaxel-resistant PCa.

miR-204和SIRT1在结直肠癌中的表达情况及临床意义

目的探讨miR-204和沉默信息调节因子1(silence information regulator 1,SIRT1)在结直肠癌组织中的表达情况及其临床价值。方法收集2018年5月至2020年6月嘉兴大学附属医院胃肠外科诊治的60例结直肠癌患者的癌组织标本和癌旁组织标本为研究对象,采用实时荧光定量聚合酶链反应检测miR-204及SIRT1的基因表达情况,Pearson分析比较miR-204与SIRT1基因的表达相关性。采用免疫组织化学SABC法检测SIRT1蛋白表达,并比较不同SIRT1蛋白表达与临床病理特征的关系。Kaplan-Meier法分析不同SIRT1蛋白表达的结直肠癌患者的生存差异。结果癌组织中miR-204基因mRNA表达显著低于癌旁组织(P<0.05),SIRT1基因mRNA表达显著高于癌旁组织(P<0.05)。癌组织中SIRT1蛋白表达阳性率显著高于癌旁组织(P<0.05)。Pearson相关分析显示miR-204与SIRT1基因mRNA在癌旁组织和癌组织中的表达均呈负相关(r=–0.647、–0.737,P<0.05)。SIRT1蛋白的表达与结直肠癌的分化水平、浸润层次、淋巴结转移与否及TNM分期相关(P<0.05),与患者的年龄、性别、肿瘤大小及肿瘤部位无关(P>0.05)。Kaplan-Meier分析显示癌组织中SIRT1阳性表达患者的生存率显著低于SIRT1阴性表达患者(χ^(2)=5.001,P=0.025)。结论结直肠癌组织中miR-204表达下调,SIRT1表达上调,二者可能通过相互影响共同促进结直肠癌的转移、侵袭,并影响患者预后。

Bioinformatic analysis reveals novel biomarkers and candidate drug compositions in prostate cancer

As prostate cancer(PC)patients do more and more genome sequencing,we can predict prognosis through individual oncogenic mutations.Although great success have been made to clarify the incidence of PC,the mechanisms was not completely understood.Recurrence and metastasis of PC remains to be resolved,and novel therapeutic targets need to be found urgently.Microarray datasets GSE6919,GSE55945 and GSE46602 about the PC tissues vs.normal organizations,were obtained from Gene Expression Omnibus.In this study,86 differentially expressed genes were determined having more important clinical significance in the process of PC.29 hub genes significantly enriched in biological processes were analyzed using Cytoscape.The function of these hub genes included the effect of cellular process,skeletal system development,cholesterol transport,regulation of protein oligomerization and cellular component biogenesis,enzyme inhibitor activity and so on.The three of these hub genes were picked out because of their relationships,which can be used as a potential target for the diagnosis and the direction of therapy.And drug predictions were designed for these candidate target molecules,providing direction for future treatment of PC.

PEG3、STMN1基因在非小细胞肺癌的表达及其与临床病理特征、血管生成相关性研究

目的 探讨父系表达基因3(PEG3)、抑微管装配蛋白1(STMN1)基因在非小细胞肺癌(NSCLC)的表达及其与临床病理特征和血管生成的关系。方法 收集宁夏医科大学总医院肿瘤医院2021年1月—2023年1月经病理证实的96例NSCLC患者癌组织及癌旁组织,采用实时荧光定量聚合酶链反应(qRT-PCR)检测和免疫组织化学检测癌组织与癌旁组织PEG3、STMN1、VEGF及CD105 mRNA的表达;比较不同临床病理特征NSCLC患者癌组织PEG3、STMN1的阳性表达率;采用Pearson法分析PEG3、STMN1与VEGF及CD105关系;采用受试者工作特征(ROC)曲线分析PEG3、STMN1对NSCLC的诊断价值。结果 与癌旁组织比较,PEG3在NSCLC组织中表达降低(P <0.05),STMN1、VEGF及CD105在NSCLC组织中表达升高(P <0.05);NSCLC组织中STMN1、VEGF及CD105阳性率分别为62.50%、69.79%和72.92%,分别高于癌旁组织5.21%、10.42%和13.54%(P <0.05),NSCLC组织中PEG3阳性率为8.33%低于癌旁组织73.96%(P <0.05);不同年龄、性别及肿瘤类型NSCLC患者的PEG3及STMN1表达水平比较,差异无统计学意义(P>0.05),不同TNM分期、淋巴结转移及分化程度NSCLC患者的PEG3及STMN1表达水平比较,差异有统计学意义(P <0.05);NSCLC组织的PEG3与STMN1、VEGF及CD105均呈负相关(P <0.05),NSCLC组织的STMN1与PEG3呈负相关(P <0.05),与VEGF及CD105均呈正相关(P <0.05);PEG3诊断NSCLC的曲线下面积为0.750(95%CI:0.453,0.936)、敏感性为73.66%(95%CI:0.650,0.937)、特异性为79.62%(95%CI:0.590,0.956);STMN1诊断NSCLC的曲线下面积为0.796(95%CI:0.540,0.942)、敏感性为80.30%(95%CI:0.744,0.978)、特异性为81.12%(95%CI:0.612,0.996);PEG3+STMN1联合诊断的曲线下面积为0.935(95%CI:0.753,0.995)、敏感性为92.33%(95%CI:0.751,0.930)、特异性为77.12%(95%CI:0.735,0.948)。结论 NSCLC组织PEG3降低、STMN1升高,与肺癌TNM分期、分化程度相关,其可以加快肿瘤血管生成,能够一定程度提高疾病诊断价值。

Effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions

Objective: To investigate the effects of eEF1A1 expression on cancer cell load and proliferation activity in lung cancer lesions. Methods: A total of 200 patients with non-small cell lung cancer who received radical operation for lung cancer in our hospital were enrolled in lung cancer group, and 78 patients with pulmonary bullae who accepted emergency surgical treatment in our hospital during the same period were enrolled in pulmonary bullae group. Differences in eEF1A1 and proliferation gene expression in lesion tissue were compared between the two groups, and the contents of serum tumor markers and angiogenesis indexes were detected. Pearson test was used to evaluate the correlation of eEF1A1 expression with tumor markers, angiogenesis indexes and proliferation genes in the lesions of patients with non-small cell lung cancer. Results: eEF1A1 expression in lesion tissue of lung cancer group was higher than that of pulmonary bullae group;serum CYFRA21-1, ProGRP, CEA, SCC Ag, HIF-1 and VEGF levels of lung cancer group were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression;LRRC3B, TUSC3 and VPS33B mRNA expression in lesion tissue of lung cancer group were lower than those of pulmonary bullae group and negatively correlated with the eEF1A1 expression whereas MACC1 and RACK1 mRNA expression were higher than those of pulmonary bullae group and positively correlated with the eEF1A1 expression. Conclusion: eEF1A1 is highly expressed in non-small cell lung cancer tissues, and the specific expression can objectively reflect the tumor load and the proliferation activity of tumor cells.

沉默SOCS1联合rAAV/PSA基因修饰的树突状细胞疫苗治疗前列腺癌的实验研究

目的研究沉默细胞因子信号抑制因子1(suppressor of cytokine signaling 1,SOCS1)基因联合rAAV/PSA基因修饰的树突状细胞(dendritic cell,DC)对前列腺癌LNCaP细胞的体外杀伤效应。方法应用重组腺相关病毒(rAAV)介导的RNA干扰技术沉默人DC的SOCS1基因的表达,行Western blot检测基因沉默效果。rAAV-shRNA-SOCS1和rAAV/PSA感染DC,将DC细胞分为Control-DC组、rAAV/PSA-DC组、SOCS1(-)+rAAV/PSA-DC组。采用系列细胞因子诱导DC成熟,诱导细胞毒性T淋巴细胞(CTL)。流式细胞术检测分析各组DC细胞表型及CTL细胞的免疫表型;ELISA法检测各组DC细胞培养上清中细胞因子白介素-10(IL-10)及IL-12 p70的分泌水平,各组CTL细胞释放γ-干扰素(IFN-γ)的水平;LDH法检测各组CTL细胞对前列腺癌LNCaP靶细胞的杀伤效率。结果rAAV-shRNA-SOCS1感染DC后,可有效下调DC的SOCS1表达水平。与对照组比较,沉默SOCS1联合rAAV/PSA基因修饰的DC细胞培养上清中细胞因子IL-12 p70的分泌水平显著增高,IL-10的分泌水平明显下降(P<0.05);该组DC表面分子CD80、CD83和CD86的表达明显上调(P<0.05)。该组DC诱导的CTL中CD8^(+)、CD8^(+)CD69^(+)T细胞的比例明显增高,而CD4^(+)T细胞、CD4^(+)CD25^(+)FoxP3^(+)Treg细胞的比例显著降低(P<0.05),IFN-γ的释放水平明显增高(P<0.05),对PSA阳性的前列腺癌靶细胞LNCaP具有更强的杀伤活性(P<0.05),且具有抗原特异性。结论沉默SOCS1联合rAAV/PSA基因修饰的DC疫苗可以产生高效而特异性的抗前列腺癌免疫效应。