摘要
目的探讨人表皮生长因子受体-2(HER-2)的表达对胃癌细胞粘附、迁移等生物学行为的影响,并探讨其表达与上皮间质转化(EMT)及Wnt/β-catenin信号通路的关系。方法利用小干扰RNA技术,在SGC-7901胃癌细胞中沉默HER-2表达;采用实时荧光定量PCR(qPCR)及Western blotting法验证HER-2的沉默效果,并将实验分为空白组、空载体组和HER-2沉默组;采用MTT比色法检测细胞粘附能力,Transwell小室法检测细胞迁移及侵袭能力;qPCR和Western blotting法检测EMT相关蛋白E-cadherin、Vimentin、Snail及Wnt/β-catenin信号通路GSK-3β、β-catenin、c-Myc的mRNA和蛋白表达情况。结果HER-2沉默组中HER-2 mRNA和蛋白表达水平低于空白组和空载体组(P<0.001),而空白组和空载体组的差异无统计学意义(P>0.05)。与空白组和空载体组比较,HER-2沉默组胃癌细胞的粘附能力、迁移能力和侵袭能力均显著下降(P<0.001)。HER-2沉默组胃癌细胞Vimentin和Snail的mRNA和蛋白表达均降低,E-cadherin的mRNA和蛋白表达升高,差异均有统计学意义(P<0.001)。与空白组和空载体组比较,HER-2沉默组胃癌细胞β-catenin和c-Myc的mRNA和蛋白表达均降低,GSK-3β的mRNA和蛋白表达升高,差异均有统计学意义(P<0.001)。结论HER-2表达影响胃癌细胞的生物学行为,其可能通过促进EMT及影响Wnt/β-catenin信号通路,从而增强胃癌细胞的粘附、迁移及侵袭能力。
Objective To study the effect of human epidermal growth factor receptor 2(HER-2)expression on cell adhesion and migration,and investigate the relationship between HER-2 expression and epithelial-mesenchymal transition(EMT),as well as Wnt/β-catenin signaling pathway.Methods Using small interfering RNA technology,HER-2 expression was silenced in SGC-7901 gastric cancer cells;The silencing effect of HER-2 was verified by real-time fluorescence PCR(qPCR)and Western blotting,and the experiments were divided into blank group,empty vector group and HER-2 silencing group.The MTT colorimetric method was used to detect cell adhesion ability,and the Transwell chamber test was used to detect cell migration and invasion ability.Western blotting and qPCR detected the expression of mRNA and protein of EMT-related proteins E-cadherin,Vimentin,Snail and Wnt/β-catenin signaling pathways related proteins of GSK-3β,β-catenin and c-Myc.Results The expression levels of HER-2 mRNA and protein in the HER-2 silencing group were lower than those in the blank group and the empty vector group(P<0.001),while the difference between the blank group and the empty vector group was not statistically significant(P>0.05).Compared with the blank group and the empty vector group,the adhesion,migration ability and invasion ability of gastric cancer cells in the HER-2 silencing group were significantly reduced(P<0.001).The expression of Vimentin and Snail in gastric cancer cells in the HER-2 silencing group reduced,and the expression of E-cadherin mRNA and protein increased,and the difference was statistically significant(P<0.001).Compared with the blank group and the empty vector group,the mRNA and protein expression ofβ-catenin and c-Myc in gastric cancer cells in the HER-2 silencing group reduced,and the expression of GSK-3βmRNA and protein increased,and the difference was statistically significant(P<0.001).Conclusion HER-2 expression affects the biological behavior of gastric cancer cells,which may enhance the adhesion,migration and invasion ability of gastric cancer cells by promoting EMT and affecting the Wnt/β-catenin signaling pathway.
作者
张雪
赵丽丽
闫文娴
冯青青
赵文文
赵文飞
井文君
魏红梅
ZHANG Xue;ZHAO Lili;YAN Wenxian;FENG Qingqing;ZHAO Wenwen;ZHAO Wenfei;JING Wenjun;WEI Hongmei(Department of Oncology,the Affiliated Qingdao Central Hospital of Qingdao University,Qingdao Cancer Hospital,Qingdao 266042,China)
出处
《临床肿瘤学杂志》
CAS
2023年第7期585-591,共7页
Chinese Clinical Oncology