靶向调控CTNNB1表达水平对胃癌细胞系SGC-7901增殖及侵袭能力的影响

Targeted regulation of CTNNB1 expression affects proliferation and invasion of gastric cancer cell line SGC-7901

目的探讨调控Wnt/CTNNB1通路对胃癌增殖和侵袭能力的影响。方法人胃癌细胞系SGC-7901,体外培养慢病毒转染的稳转SGC-7901,分成CTNNB1-shRNA组、shRNA组与对照组。qRT-PCR和免疫印迹检测稳转细胞系中AKT、CTNNB1、Wnt2、Cyp19A1的表达,CCK-8测定不同细胞系增殖能力,划痕试验测定胃癌细胞系的转移能力,Transwell小室测定胃癌细胞系侵袭能力,集落形成试验测定胃癌细胞系的集落形成能力。结果AKT、CTNNB1、Wnt2、Cyp19A1 mRNA和蛋白在CTNNB1-shRNA组表达最低(P<0.05),在shRNA组和对照组中表达差异无统计学意义(P>0.05);24、48、72 h细胞增殖水平在CTNNB1-shRNA组细胞最低(P<0.05),shRNA组和对照组细胞增殖差异无统计学意义(P>0.05);划痕后24 h CTNNB1-shRNA组24 h迁移细胞率最低(P<0.05),shRNA组与对照组迁移细胞率差异无统计意义(P>0.05);CTNNB1-shRNA组侵袭细胞数量和集落形成数量最低(P<0.05),shRNA组与对照组侵袭细胞数量差异无统计意义(P>0.05)。结论通过下调Wnt/CTNNB1通路能引起该通路相关下游因子在胃癌细胞中低表达,从而影响胃癌增殖和侵袭能力,有望成为胃癌治疗新靶点。

Objective To investigate the effect of regulating the Wnt/CTNNB1 pathway on proliferation and invasion of gastric cancer.Methods Human gastric cancer cell line SGC-7901 was used in this study.The SGC-7901 were cultured in vitro,stably-transfected with lentivirus,and divided into the CTNNB1-shRNA group,shRNA group and control group.qRT-PCR and Western blotting were used to detect the expression of AKT,CTNNB1,Wnt2 and Cyp19A1 in the stably transfected cell line.CCK-8 assay was used to examine the proliferation ability of different cell groups.Scratch test was used to assess the migration ability of gastric cancer cell lines.Transwell chamber assay was used to measure the invasion of gastric cancer cell lines.Capacity,colony-forming assays determine the colony-forming capability of gastric cancer cell lines.Results The mRNA and protein expression levels of AKT,CTNNB1,Wnt2,and Cyp19A1 were the lowest in the CTNNB1-shRNA group(P<0.05),and did not differ significantly between the shRNA group and control group(P>0.05).At 24,48,and 72 h,the cell proliferation level was the lowest in the CTNNB1-shRNA group(P<0.05),and did not differ significantly between the shRNA group and the control group(P>0.05)either.At 24 h post-scratching,the 24 h migration rate was the lowest in the CTNNB1-shRNA group(P<0.05),and did not differ between the shRNA group and the control group(P>0.05);the CTNNB1-shRNA group showed lowest cell migration and colony formation(P<0.05).The number of migrating cells did not differ significantly between the shRNA group and the control group(P>0.05).Conclusion Down-regulating the Wnt/CTNNB1 pathway can lead to low expression of downstream signaling molecules of this pathway in gastric cancer cell lines and hence interference with proliferation and invasion of gastric cancer.This offers clue to new target for treatment of gastric cancer.