背景随着医医疗水平的提高,胃癌的早期诊断及治疗取得了明显成效,但患者的死亡率仍居高不下,且预后较差,侵袭与转移是影响患者预后的重要因素.因此,探究胃癌的发病机制,抑制胃癌细胞的侵袭转移,改善患者预后意义重大.乙酰肝素酶(heparan...背景随着医医疗水平的提高,胃癌的早期诊断及治疗取得了明显成效,但患者的死亡率仍居高不下,且预后较差,侵袭与转移是影响患者预后的重要因素.因此,探究胃癌的发病机制,抑制胃癌细胞的侵袭转移,改善患者预后意义重大.乙酰肝素酶(heparanase,HPSE)是一种内源性糖苷内切酶,有研究发现,期在胃癌组织中的表达水平明显升高.但关于乙酰肝素酶2(heparanase,HPSE2)在胃癌中的相关作用及其作用机制尚不明确.本研究旨在HPSE2在胃癌中的相关作用及其作用机制.目的分析HPSE2通过抑制核转录因子κB(NF-κB)、wnt基因/β-连环蛋白(Wnt/β-catenin)信号通路调控胃癌细胞恶性生物学行为的机制.方法收集2019-09/2021-04期间在本院进行手术切除治疗的74例胃癌患者的胃癌组织及癌旁正常组织,测定HPSE2的表达水平.将人胃癌细胞系MKN-28细胞进行传代培养并转染,获得HPSE2过表达组及对照组.测定各组细胞增殖能力、细胞凋亡率、细胞侵袭及迁移能力,并测定细胞中HPSE2、上皮型钙黏蛋白(epithelial cadherin,E-cadherin)、神经型钙黏蛋白(neuralcadherin,N-cadherin)、波形蛋白(Vimentin)、转化生长因子-β1(transforming growth factor-β1,T G F-β1)、磷酸化核转录因子-κBp65(n u c l e a r transcriptionfactorκBp65,p-NF-κBp65)、β-catenin表达水平变化.结果与癌旁正常组织对比,胃癌组织中HPSE2表达水平明显降低(P<0.01).与对照组比较,HPSE2过表达组HPSE2表达水平明显升高(P<0.01).两组1d时细胞增殖能力无明显差异(P>0.05);与2 d与对照组比较,H P S E2过表达组细胞增殖能力明显降低(P<0.05或P<0.01).与对照组比较,HPSE2过表达组细胞凋亡率及Bax、E-cadherin表达水平均明显升高,细胞侵袭、迁移能力及Bax、Suivivin、N-cadherin、Vimentin、TGF-β1、p-NF-κBp65、β-catenin表达水平均明显降低(P<0.01).结论上调HPSE2表达能够明显抑制细胞增殖、侵袭及迁移,抑制细胞发生EMT,促进细胞凋亡,其可能是通过抑制NF-κB及Wnt/β-catenin信号通路的激活实现.展开更多
Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such inf...Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.展开更多
AIM: To investigate member 3a of Wingless-type MMTV integration site family (Wnt3a) expression in cancerous and surrounding tissues and the relationship between clinicopathologic features of hepatocellular carcinoma (...AIM: To investigate member 3a of Wingless-type MMTV integration site family (Wnt3a) expression in cancerous and surrounding tissues and the relationship between clinicopathologic features of hepatocellular carcinoma (HCC) and Wnt3a expression.METHODS: Wnt3a expression and cellular distribution and clinicopathologic characteristics in cancerous tissue and matched surrounding tissues were analyzed in 80 HCC patients from January 2006 to August 2008 by tissue microarrays and immunohistochemistry. The overall and disease-free survival rates were estimated using the Kaplan-Meier method and compared with the log-rank test. The prognostic analysis was carried out with univariate and multivariate Cox regressions models.RESULTS: The incidence of oncogenic Wnt3a expression in the cancerous group was up to 96.25% (77 of 80), which was significantly higher (χ<sup>2</sup> = 48.818, P < 0.001) than that in the surrounding group (46.25%, 37 of 80). Brown Wnt3a staining gradually increased with clinical staging that showed very strong staining in advanced HCC. The clinicopathologic features of high Wnt3a expression in HCC were related to poorly-differentiated grade (χ<sup>2</sup> = 20.211, P < 0.001), liver cirrhosis (χ<sup>2</sup> = 8.467, P < 0.004), hepatitis B virus (HBV) infection (χ<sup>2</sup> = 12.957, P < 0.001), higher tumor-node-metastasis stage (χ<sup>2</sup> = 22.960, P < 0.001), and 5-year survival rate (χ<sup>2</sup> = 15.469, P < 0.001).CONCLUSION: Oncogenic Wnt3a expression associated with HBV infection and cirrhotic liver might be an independent prognostic factor for HCC.展开更多
Multiple lines of evidence indicate that Wnt/β-catenin signaling plays a fundamental role in colorectal cancer (CRC) initiation and progression. Recent genome-wide data have confirmed that in CRC this path...Multiple lines of evidence indicate that Wnt/β-catenin signaling plays a fundamental role in colorectal cancer (CRC) initiation and progression. Recent genome-wide data have confirmed that in CRC this pathway is one of the most frequently modified by genetic or epigenetic alterations affecting almost 90% of Wnt/β-catenin gene members. A major challenge is thus learning how the corrupted coordination of this pathway is tied to other signalings to enhance cell growth. Peroxisome proliferator activated receptor γ (PPARγ) is emerging as a growth-limiting and differentiation-promoting factor. In tumorigenesis it exerts a tumor suppressor role and is potentially linked with the Wnt/β-catenin pathway. Based on these results, the identification of new selective PPARγ modulators with inhibitory effects on the Wnt/β-catenin pathway is becoming an interesting perspective. Should, in fact, these molecules display such properties, new research avenues would be opened aimed at developing new molecular targeted drugs. Herein, we review the basic principles and present new hypotheses underlying the crosstalk between Wnt/β-catenin and PPARγ signaling. Furthermore, we discuss the advances in our understanding as to how their altered regulation can culminate in colon cancer and the efforts aimed at designing novel PPARγ agonists endowed with Wnt/β-catenin inhibitory effects to be used as therapeutic and/or preventive agents.展开更多
AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms (SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 pa...AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms (SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 paraffin-embedded archived intestinal-type gastric cancer samples and 129 biopsies obtained by superior endoscopy from chronic gastritis patients. Gastric cancer samples were classified according the degree of carcinoma differentiation. Genomic DNA was extracted from tissues, and the two SNPs of TCF7L2 gene (rs12255372 and rs7903146) were genotyped by polymerase chain reaction-restriction fragment length polymorphism reactions. Multiple regression analysis with adjustments for age and gender were performed and best-fitting models of inheritance were determined. Statistic powers were post-hoc calculated.RESULTS: After adjusting for age and sex the TCF7L2 rs7903146 TT genotype was associated with gastric cancer risk under the recessive genetic model (OR = 3.11, 95%CI: 1.22-7.92, P = 0.017). We further investigated the distribution of rs12255372 and rs7903146 genotypes according gastric cancer stratified by degree of differentiation, and we observed that carriers of rs7903146 T allele (CT + TT vs CC) had a significantly increased risk of moderate/well differentiated gastric cancer (dominant model, OR = 2.55, 95%CI: 1.35-4.80, P = 0.004), whereas the rs7903146 TT genotype was associated with poorly differentiated gastric cancer in the recessive model (OR = 3.65, 95%CI: 1.25-10.62, P = 0.018). We did not find association between rs12255372 SNP and the susceptibility of developing gastric cancer.CONCLUSION: TCF7L2 rs7903146 polymorphism is associated with gastric cancer risk in the Venezuelan population, and could be related to determine the degree of differentiation of tumor cells.展开更多
目的:探讨芪玉三龙方对荷肺癌小鼠的抑瘤效果和对Wnt/β-catenin信号通路下游靶基因c-myc、cyclin D1、Axin2表达的影响。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,随机分为模型组,芪玉三龙方低、中、高剂量组,化疗组,...目的:探讨芪玉三龙方对荷肺癌小鼠的抑瘤效果和对Wnt/β-catenin信号通路下游靶基因c-myc、cyclin D1、Axin2表达的影响。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,随机分为模型组,芪玉三龙方低、中、高剂量组,化疗组,联合组,每组8只;造模第2天开始予以芪玉三龙方低、中、高剂量组小鼠分别按20.12、40.24、80.48g·kg-1·d-1灌胃给药,化疗组按照0.4m L顺铂腹腔注射给药,联合组以高剂量中药灌胃和顺铂腹腔注射联合给药,模型组按照等量0.9%氯化钠溶液给药,连续给药21d;称取瘤质量,计算抑瘤率;RT-q PCR检测肿瘤组织c-myc、cyclin D1、Axin2 m RNA的转录水平。结果:与模型组和低剂量组比较,芪玉三龙方中、高剂量组和化疗组c-myc m RNA表达显著降低(P<0.01)。与模型组比较,各用药组cyclin D1 m RNA表达显著降低(P<0.05,P<0.01);与低剂量组比较,中剂量组、化疗组表达显著降低(P<0.01)。与模型组比较,各用药组Axin2 m RNA表达显著降低(P<0.01);与低剂量组、化疗组比较,芪玉三龙方中、高剂量组表达显著降低(P<0.01)。结论:芪玉三龙方对肺癌移植瘤具有一定的抑制作用,其抑瘤机制可能与芪玉三龙方调节肺癌组织c-myc m RNA、cyclin D1 m RNA、Axin2 m RNA的转录水平有关。芪玉三龙方高剂量抑瘤效应较优,量效关系明显,其与化疗药合用具有协同效应。展开更多
Cancer stem cells(CSC) are thought to be malignant cells that have the capacity to initiate and maintain tumor growth and survival. Studies have described CSC in various gastrointestinal neoplasms such as colon, pancr...Cancer stem cells(CSC) are thought to be malignant cells that have the capacity to initiate and maintain tumor growth and survival. Studies have described CSC in various gastrointestinal neoplasms such as colon, pancreas and liver and gastroesophageal tumors. The mechanism by which CSC develop remains unclear. Several studies have explored the role of dysregulation of the Wnt/β-catenin, transformation growth factor-beta and hedhog pathways in generation of CSC. In this review, we discuss the various molecular abnormalities that may be related to formation of CSC in gastrointestinal malignancies, strategies to identify CSC and therapeutic strategies that are based on these concepts. Identification and targeting CSC is an intriguing area and may provide a new therapeutic option for patients with cancer including gastrointestinal malignancies. Although great progress has been made, many issues need to be addressed. Precise targeting of CSC will require precise isolation and characterization of those cells. This field is also evolving but further research is needed to identify markers that are specific for CSC.Although the application of this field has not entered the clinic yet, there continues to be significant optimism about its potential utility in overcoming cancer resistance and curing patients with cancer.展开更多
文摘背景随着医医疗水平的提高,胃癌的早期诊断及治疗取得了明显成效,但患者的死亡率仍居高不下,且预后较差,侵袭与转移是影响患者预后的重要因素.因此,探究胃癌的发病机制,抑制胃癌细胞的侵袭转移,改善患者预后意义重大.乙酰肝素酶(heparanase,HPSE)是一种内源性糖苷内切酶,有研究发现,期在胃癌组织中的表达水平明显升高.但关于乙酰肝素酶2(heparanase,HPSE2)在胃癌中的相关作用及其作用机制尚不明确.本研究旨在HPSE2在胃癌中的相关作用及其作用机制.目的分析HPSE2通过抑制核转录因子κB(NF-κB)、wnt基因/β-连环蛋白(Wnt/β-catenin)信号通路调控胃癌细胞恶性生物学行为的机制.方法收集2019-09/2021-04期间在本院进行手术切除治疗的74例胃癌患者的胃癌组织及癌旁正常组织,测定HPSE2的表达水平.将人胃癌细胞系MKN-28细胞进行传代培养并转染,获得HPSE2过表达组及对照组.测定各组细胞增殖能力、细胞凋亡率、细胞侵袭及迁移能力,并测定细胞中HPSE2、上皮型钙黏蛋白(epithelial cadherin,E-cadherin)、神经型钙黏蛋白(neuralcadherin,N-cadherin)、波形蛋白(Vimentin)、转化生长因子-β1(transforming growth factor-β1,T G F-β1)、磷酸化核转录因子-κBp65(n u c l e a r transcriptionfactorκBp65,p-NF-κBp65)、β-catenin表达水平变化.结果与癌旁正常组织对比,胃癌组织中HPSE2表达水平明显降低(P<0.01).与对照组比较,HPSE2过表达组HPSE2表达水平明显升高(P<0.01).两组1d时细胞增殖能力无明显差异(P>0.05);与2 d与对照组比较,H P S E2过表达组细胞增殖能力明显降低(P<0.05或P<0.01).与对照组比较,HPSE2过表达组细胞凋亡率及Bax、E-cadherin表达水平均明显升高,细胞侵袭、迁移能力及Bax、Suivivin、N-cadherin、Vimentin、TGF-β1、p-NF-κBp65、β-catenin表达水平均明显降低(P<0.01).结论上调HPSE2表达能够明显抑制细胞增殖、侵袭及迁移,抑制细胞发生EMT,促进细胞凋亡,其可能是通过抑制NF-κB及Wnt/β-catenin信号通路的激活实现.
基金We thank the Experimental Poultry Farm of the Poultry Institute,Chinese Academy of Agricultural Sciences,for providing experimental materialsThis work was supported by the Key Research and Development Program of China(2017YFE0108000)+1 种基金the National Natural Science Foundation of China(31872341,31572390)the High-Level Talent Support Program of Yangzhou University,China.
文摘Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.
基金Supported by the International S&T Cooperation Program,No.2013DFA32150 of China
文摘AIM: To investigate member 3a of Wingless-type MMTV integration site family (Wnt3a) expression in cancerous and surrounding tissues and the relationship between clinicopathologic features of hepatocellular carcinoma (HCC) and Wnt3a expression.METHODS: Wnt3a expression and cellular distribution and clinicopathologic characteristics in cancerous tissue and matched surrounding tissues were analyzed in 80 HCC patients from January 2006 to August 2008 by tissue microarrays and immunohistochemistry. The overall and disease-free survival rates were estimated using the Kaplan-Meier method and compared with the log-rank test. The prognostic analysis was carried out with univariate and multivariate Cox regressions models.RESULTS: The incidence of oncogenic Wnt3a expression in the cancerous group was up to 96.25% (77 of 80), which was significantly higher (χ<sup>2</sup> = 48.818, P < 0.001) than that in the surrounding group (46.25%, 37 of 80). Brown Wnt3a staining gradually increased with clinical staging that showed very strong staining in advanced HCC. The clinicopathologic features of high Wnt3a expression in HCC were related to poorly-differentiated grade (χ<sup>2</sup> = 20.211, P < 0.001), liver cirrhosis (χ<sup>2</sup> = 8.467, P < 0.004), hepatitis B virus (HBV) infection (χ<sup>2</sup> = 12.957, P < 0.001), higher tumor-node-metastasis stage (χ<sup>2</sup> = 22.960, P < 0.001), and 5-year survival rate (χ<sup>2</sup> = 15.469, P < 0.001).CONCLUSION: Oncogenic Wnt3a expression associated with HBV infection and cirrhotic liver might be an independent prognostic factor for HCC.
基金Supported by Ministero dell’Istruzione,Università e Ricerca,MIUR-PRIN 2010-2011,No.prot.2010W7YRLZ_003
文摘Multiple lines of evidence indicate that Wnt/β-catenin signaling plays a fundamental role in colorectal cancer (CRC) initiation and progression. Recent genome-wide data have confirmed that in CRC this pathway is one of the most frequently modified by genetic or epigenetic alterations affecting almost 90% of Wnt/β-catenin gene members. A major challenge is thus learning how the corrupted coordination of this pathway is tied to other signalings to enhance cell growth. Peroxisome proliferator activated receptor γ (PPARγ) is emerging as a growth-limiting and differentiation-promoting factor. In tumorigenesis it exerts a tumor suppressor role and is potentially linked with the Wnt/β-catenin pathway. Based on these results, the identification of new selective PPARγ modulators with inhibitory effects on the Wnt/β-catenin pathway is becoming an interesting perspective. Should, in fact, these molecules display such properties, new research avenues would be opened aimed at developing new molecular targeted drugs. Herein, we review the basic principles and present new hypotheses underlying the crosstalk between Wnt/β-catenin and PPARγ signaling. Furthermore, we discuss the advances in our understanding as to how their altered regulation can culminate in colon cancer and the efforts aimed at designing novel PPARγ agonists endowed with Wnt/β-catenin inhibitory effects to be used as therapeutic and/or preventive agents.
文摘AIM: To explore the association between TCF7L2 rs12255372 and rs7903146 single nucleotide polymorphisms (SNPs) and gastric cancer risk in Venezuelan patients.METHODS: We performed a case-control study including 122 paraffin-embedded archived intestinal-type gastric cancer samples and 129 biopsies obtained by superior endoscopy from chronic gastritis patients. Gastric cancer samples were classified according the degree of carcinoma differentiation. Genomic DNA was extracted from tissues, and the two SNPs of TCF7L2 gene (rs12255372 and rs7903146) were genotyped by polymerase chain reaction-restriction fragment length polymorphism reactions. Multiple regression analysis with adjustments for age and gender were performed and best-fitting models of inheritance were determined. Statistic powers were post-hoc calculated.RESULTS: After adjusting for age and sex the TCF7L2 rs7903146 TT genotype was associated with gastric cancer risk under the recessive genetic model (OR = 3.11, 95%CI: 1.22-7.92, P = 0.017). We further investigated the distribution of rs12255372 and rs7903146 genotypes according gastric cancer stratified by degree of differentiation, and we observed that carriers of rs7903146 T allele (CT + TT vs CC) had a significantly increased risk of moderate/well differentiated gastric cancer (dominant model, OR = 2.55, 95%CI: 1.35-4.80, P = 0.004), whereas the rs7903146 TT genotype was associated with poorly differentiated gastric cancer in the recessive model (OR = 3.65, 95%CI: 1.25-10.62, P = 0.018). We did not find association between rs12255372 SNP and the susceptibility of developing gastric cancer.CONCLUSION: TCF7L2 rs7903146 polymorphism is associated with gastric cancer risk in the Venezuelan population, and could be related to determine the degree of differentiation of tumor cells.
文摘目的:探讨芪玉三龙方对荷肺癌小鼠的抑瘤效果和对Wnt/β-catenin信号通路下游靶基因c-myc、cyclin D1、Axin2表达的影响。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,随机分为模型组,芪玉三龙方低、中、高剂量组,化疗组,联合组,每组8只;造模第2天开始予以芪玉三龙方低、中、高剂量组小鼠分别按20.12、40.24、80.48g·kg-1·d-1灌胃给药,化疗组按照0.4m L顺铂腹腔注射给药,联合组以高剂量中药灌胃和顺铂腹腔注射联合给药,模型组按照等量0.9%氯化钠溶液给药,连续给药21d;称取瘤质量,计算抑瘤率;RT-q PCR检测肿瘤组织c-myc、cyclin D1、Axin2 m RNA的转录水平。结果:与模型组和低剂量组比较,芪玉三龙方中、高剂量组和化疗组c-myc m RNA表达显著降低(P<0.01)。与模型组比较,各用药组cyclin D1 m RNA表达显著降低(P<0.05,P<0.01);与低剂量组比较,中剂量组、化疗组表达显著降低(P<0.01)。与模型组比较,各用药组Axin2 m RNA表达显著降低(P<0.01);与低剂量组、化疗组比较,芪玉三龙方中、高剂量组表达显著降低(P<0.01)。结论:芪玉三龙方对肺癌移植瘤具有一定的抑制作用,其抑瘤机制可能与芪玉三龙方调节肺癌组织c-myc m RNA、cyclin D1 m RNA、Axin2 m RNA的转录水平有关。芪玉三龙方高剂量抑瘤效应较优,量效关系明显,其与化疗药合用具有协同效应。
文摘Cancer stem cells(CSC) are thought to be malignant cells that have the capacity to initiate and maintain tumor growth and survival. Studies have described CSC in various gastrointestinal neoplasms such as colon, pancreas and liver and gastroesophageal tumors. The mechanism by which CSC develop remains unclear. Several studies have explored the role of dysregulation of the Wnt/β-catenin, transformation growth factor-beta and hedhog pathways in generation of CSC. In this review, we discuss the various molecular abnormalities that may be related to formation of CSC in gastrointestinal malignancies, strategies to identify CSC and therapeutic strategies that are based on these concepts. Identification and targeting CSC is an intriguing area and may provide a new therapeutic option for patients with cancer including gastrointestinal malignancies. Although great progress has been made, many issues need to be addressed. Precise targeting of CSC will require precise isolation and characterization of those cells. This field is also evolving but further research is needed to identify markers that are specific for CSC.Although the application of this field has not entered the clinic yet, there continues to be significant optimism about its potential utility in overcoming cancer resistance and curing patients with cancer.
