摘要
目的:探讨长基因间非编码RNA 00467(LINC00467)是否靶向miR-495-3p调控胃癌细胞HGC-27的增殖和迁移。方法:采用实时定量PCR(RT-qPCR)技术检测HGC-27细胞以及人胃黏膜上皮细胞GES-1中LINC00467和miR-495-3p表达量。采用双荧光素酶报告实验和RT-qPCR确定LINC00467和miR-495-3p之间的靶向调控关系。将HGC-27细胞分为si-NC组、si-LINC00467组、si-LINC00467+anti-miR-NC组、si-LINC00467+anti-miR-495-3p组。采用CCK-8法、平板克隆实验检测细胞增殖;划痕愈合实验检测细胞迁移;免疫印迹法分析上皮细胞钙粘蛋白(E-cadherin)和神经钙黏素(N-cadherin)表达。结果:与GES-1细胞相比,HGC-27细胞中LINC00467表达量显著升高(P<0.05),miR-495-3p表达量显著降低(P<0.05)。LINC00467与miR-495-3p直接结合。与si-NC组比较,si-LINC00467组HGC-27细胞存活率、克隆形成数、划痕愈合率、N-cadherin蛋白表达量均显著降低(P<0.05),E-cadherin蛋白表达量、miR-495-3p表达量显著升高(P<0.05)。与si-LINC00467+anti-miR-NC组比较,si-LINC00467+anti-miR-495-3p组HGC-27细胞存活率、克隆形成数、划痕愈合率、N-cadherin蛋白表达量均显著升高(P<0.05),E-cadherin蛋白表达量显著降低(P<0.05)。结论:干扰LINC00467通过上调miR-495-3p表达能够抑制胃癌细胞HGC-27增殖和迁移。
Objective:To study whether long intergenic non-coding RNA 00467(LINC00467)targets miR-495-3p to regulate the proliferation and migration of gastric cancer cell HGC-27.Methods:The expression of LINC00467 and miR-495-3p in HGC-27 cells and human gastric epithelial cells GES-1 was detected by real-time quantitative PCR(RT-qPCR).The dual luciferase reporter experiment and RT-qPCR were used to determine the targeted regulation relationship between LINC00467 and miR-495-3p HGC-27 cells were divided into si-NC group,si-LINC00467 group,si-LINC00467+anti-miR-NC group,si-LINC00467+anti-miR-495-3p group.The Cell Counting Kit(CCK-8)method and plate cloning experiment were applied to detect cell proliferation;the scratch healing experiment was used to detect cell migration;the expression of E-cadherin and N-cadherin in epithelial cells was analyzed by western blotting.Results:Compared with GES-1 cells,the expression of LINC00467 in HGC-27 cells was significantly increased(P<0.05),whereas the expression of miR-495-3p was significantly decreased(P<0.05).LINC00467 directly binds to miR-495-3p Compared with the si-NC group,the cell survival rate,number of clone cell numbers,scratch healing rate,N-cadherin protein expression of HGC-27 cells in the si-LINC00467 group were significantly reduced(P<0.05),E-cadherin protein expression and miR-495-3p expression were significantly increased(P<0.05).Compared with the si-LINC00467+anti-miR-NC group,the cell survival rate,number of clone cell numbers,scratch healing rate,and N-cadherin protein expression of HGC-27 cells in the si-LINC00467+anti-miR-495-3p group were significantly increased(P<0.05),E-cadherin protein expression was significantly reduced(P<0.05).Conclusion:Interfering with LINC00467 can inhibit the proliferation and migration of gastric cancer cells HGC-27 by up-regulating the expression of miR-495-3p.
作者
潘攀
顾永娟
徐丽贤
陈亚楠
沈冬
PAN Pan;GU Yong-juan;XU Li-xian;CHEN Ya-nan;SHEN Dong(Department of Oncology,Zhangjiagang Hospital Affiliated to Suzhou University,Jiangsu 215600,China)
出处
《中国现代普通外科进展》
CAS
2023年第4期258-262,共5页
Chinese Journal of Current Advances in General Surgery
基金
苏州市“科教兴卫”青年科技项目(KJXW2016052)。
