下调claudin-1对胆囊癌细胞SGC996生物学行为的影响
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摘要
目的探讨下调claudin-1对胆囊癌细胞生物学行为的影响。方法通过慢病毒LV-CLDN1-RNAi感染下调claudin-1在胆囊癌细胞SGC996中的表达。检测细胞增值、细胞周期、细胞凋亡、细胞侵袭和凋亡相关基因Bax及Bcl-2蛋白表达水平,分析下调claudin-1在胆囊癌细胞中的表达后对其增殖、凋亡及侵袭等生物学行为的影响。结果感染LV-CLDN1-RNAi后SGC996增殖无明显变化,被阻滞的G1期细胞明显增多(P=0.003 7),S期细胞明显减少(P=0.0021),细胞凋亡比例增加(P <0.021 1)。下调claudin-1表达能抑制胆囊癌细胞侵袭;促凋亡基因Bax的蛋白表达增加,抑制凋亡基因Bcl-2的蛋白表达降低。结论下调claudin-1表达能有效促进胆囊癌细胞的凋亡,抑制其侵袭,提示claudin-1在胆囊癌的复发和转移中可能会发挥作用。
Objective To investigate the effects of claudin-1 downregulation on the physiological processes of gallbladder cancer cells.Methods The gallbladder cancer cell line SGC996 was transfected with lentiviral LV-CLDN1-RNAi to downregulate claudin-1 expression, and the consequences on cell proliferation, cell cycle, apoptosis, and cell invasion were assessed. Results After transfection with LV-CLDN1-RNAi, the results showed no significant changes in the proliferation of the SGC996 cells. However, the amount of cells arrested in the G1 phase increased significantly(P=0.003 7), whereas the amount of cells arrested in the S phase was significantly reduced(P=0.002 1). Down-regulated expression of claudin-1 also increased the ratio of cell apoptosis(P<0.021 1), which was confirmed by the results of western blot analysis, showing that the levels of the pro-apoptosis Bax protein and anti-apoptosis Bcl-2 protein were increased and decreased, respectively. Finally, a Transwell assay indicated that claudin-1 down-regulation inhibited cell invasion. Conclusion Reducing the claudin-1 expression level could promote the apoptosis of gallbladder cancer cells and inhibit cell invasion, suggesting that claudin-1 may play a role in the recurrence and metastasis of gallbladder cancer.
Objective To investigate the effects of claudin-1 downregulation on the physiological processes of gallbladder cancer cells.Methods The gallbladder cancer cell line SGC996 was transfected with lentiviral LV-CLDN1-RNAi to downregulate claudin-1 expression, and the consequences on cell proliferation, cell cycle, apoptosis, and cell invasion were assessed. Results After transfection with LV-CLDN1-RNAi, the results showed no significant changes in the proliferation of the SGC996 cells. However, the amount of cells arrested in the G1 phase increased significantly(P=0.003 7), whereas the amount of cells arrested in the S phase was significantly reduced(P=0.002 1). Down-regulated expression of claudin-1 also increased the ratio of cell apoptosis(P<0.021 1), which was confirmed by the results of western blot analysis, showing that the levels of the pro-apoptosis Bax protein and anti-apoptosis Bcl-2 protein were increased and decreased, respectively. Finally, a Transwell assay indicated that claudin-1 down-regulation inhibited cell invasion. Conclusion Reducing the claudin-1 expression level could promote the apoptosis of gallbladder cancer cells and inhibit cell invasion, suggesting that claudin-1 may play a role in the recurrence and metastasis of gallbladder cancer.
引文
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[14]BouchagierKA,AssimakopoulosSF,KaraviasDD,etal.Expression of claudins-1,-4,-5,-7 and occludin in hepatocellular carcinoma and their relation with classic clinicopathological features and patients'survival[J]. In Vivo,2014,28(3):315-326.
[15]Matsuoka T,Mitomi H,Fukui N,et al. Cluster analysis of claudin-l and 4,E-eadhefin,andβ-catenin expression in colorectal cancers[J]. Surg Oncol,2011,103(7):674-686.
[16]Süren D,Y?ld?r?m M,Kaya V,et al. Loss of tight junction proteins(Claudin 1, 4, and 7)correlates with aggressive behavior in colorectal carcinoma[J]. Med Sci Monit,2014,20:1255.
[17]郭兴军,王敏,江建新,等. ZO-1和Claudin-1在胆管癌中的表达及其意义[J].中华实验外科杂志,2014,31(8):1739-1741.
[18]Atsushi Shiozaki,Hiroki Shimizu,Daisuke Ichikawa,et al. Claudin1 mediates tumor necrosis factor alpha-induced cell migration in human gastric cancer cells[J]. World J Gastroenterol,2014,20(47):17863-17876.
[19]Shiozaki A,Bai XH,Shen-Tu G,et al. Claudin 1 mediates TNF-α-induced gene expression and cell migration in human lung carcinoma cells[J]. PLoS One,2012,7:e38049.
[20]Jilian L Pope,Rizwan Ahmad,Ajaz A Bhat,et al. Claudin-1overexpression in intestinal epithelial cells enhances susceptibility to adenamatous polyposis coli-mediated colon tumorigenesis[J].Molecular Cancer,2014,13:167.
[2]Cui J,Liu Y,Wang X. The Roles of Glycodelin in Cancer Development and Progression[J]. Frontiers in Immunology,2017,8:1685.
[3]Xu W,Yang Z,Lu N. A new role for the PI3K/Akt signaling pathway in the epithelial-mesenchymal transition[J]. Cell Adhesion&Migration,2015,9(4):317-324.
[4]金浩,刘会春,于东红,等. Claudin-1在胆囊癌中的表达及其意义[J].蚌埠医学院学报,2012,37(1):22-24.
[5]Xu S,Zhan M,Wang J. Epithelial-to-mesenchymal transition in gallbladder cancer:from clinical evidence to cellular regulatory networks[J]. Cell Death Discovery,2017,3:17069.
[6]Lu Y,Jing J,Sun L,et al. Expression of claudin-11,-23 in different gastric tissues and its relationship with the risk and prognosis of gastric cancer[J]. PLoS One,2017,12(3):e0174476.
[7]Meric-Bernstam F,Johnson A,Holla V,et al. A decision support framework for genomically informed investigational cancer therapy[J].J Natl Cancer Inst,2015,107(7):pii:djv098.
[8]Gruber R,B?rnchen C,Rose K,et al. Diverse regulation of claudin-1 and claudin-4 in atopic dermatitis[J]. Am J Pathol,2015,185(10):2777-2789.
[9]Hahn-Str?mberg V,Askari S,Befekadu R,et al. Polymorphisms in the CLDN1 and CLDN7 genes are related to differentiation and tumor stage in colon carcinoma[J]. APMIS,2014,122(7):636-642.
[10]Luu HN,Lin HY,S?rensen KD,et al. miRNAs associated with prostate cancer risk and progression[J]. BMC Urology,2017,17:18.
[11]Suren D,Yildirim M,Kaya V,et al. Expression patterns of claudins 1,4, and 7 and their prognostic significance in nasopharyngeal carcinoma[J]. J BUON,2015,20(1):212-217.
[12]Sappayatosok K,Phattarataratip E. Overexpression of Claudin-1is Associated with Advanced Clinical Stage and Invasive Pathologic Characteristics of Oral Squamous Cell Carcinoma[J]. Head and Neck Pathol,2015,9:173-180.
[13]Holczbauerá,Gy?ngy?si B,Lotz G,et al,Increased expression of claudin-1 and claudin-7 in liver cirrhosis and hepatocellular carcinoma[J]. Pathol Oncol Res,2014,20(3):493-502.
[14]BouchagierKA,AssimakopoulosSF,KaraviasDD,etal.Expression of claudins-1,-4,-5,-7 and occludin in hepatocellular carcinoma and their relation with classic clinicopathological features and patients'survival[J]. In Vivo,2014,28(3):315-326.
[15]Matsuoka T,Mitomi H,Fukui N,et al. Cluster analysis of claudin-l and 4,E-eadhefin,andβ-catenin expression in colorectal cancers[J]. Surg Oncol,2011,103(7):674-686.
[16]Süren D,Y?ld?r?m M,Kaya V,et al. Loss of tight junction proteins(Claudin 1, 4, and 7)correlates with aggressive behavior in colorectal carcinoma[J]. Med Sci Monit,2014,20:1255.
[17]郭兴军,王敏,江建新,等. ZO-1和Claudin-1在胆管癌中的表达及其意义[J].中华实验外科杂志,2014,31(8):1739-1741.
[18]Atsushi Shiozaki,Hiroki Shimizu,Daisuke Ichikawa,et al. Claudin1 mediates tumor necrosis factor alpha-induced cell migration in human gastric cancer cells[J]. World J Gastroenterol,2014,20(47):17863-17876.
[19]Shiozaki A,Bai XH,Shen-Tu G,et al. Claudin 1 mediates TNF-α-induced gene expression and cell migration in human lung carcinoma cells[J]. PLoS One,2012,7:e38049.
[20]Jilian L Pope,Rizwan Ahmad,Ajaz A Bhat,et al. Claudin-1overexpression in intestinal epithelial cells enhances susceptibility to adenamatous polyposis coli-mediated colon tumorigenesis[J].Molecular Cancer,2014,13:167.