长链非编码RNA RP11-497E19.1对胃癌细胞增殖和侵袭的影响及机制实验研究

Effect of long non-coding RNA RP11-497E19.1 on proliferation and invasion of gastric cancer cells and its mechanism

目的:探讨长链非编码RNA(lncRNA)RP11-497E19.1对胃癌细胞增殖和侵袭的影响及其机制。方法:实时荧光定量PCR(RT-qPCR)检测胃癌HS-746T、BGC823、NCI-N87、SGC7901、AGS细胞和永生化胃上皮细胞GES-1中RP11-497E19.1表达水平,筛选表达最高的细胞进行后续实验。将HS-746T细胞分为si-NC组和si-RP11-497E19.1组,分别转染阴性对照寡核苷酸或RP11-497E19.1小干扰RNA。集落形成实验和Transwell实验分析转染HS-746T细胞的增殖、侵袭能力。双荧光素酶报告基因实验验证RP11-497E19.1与微小RNA(miR)-545-5p的靶向关系。RT-qPCR检测转染HS-746T细胞miR-545-5p的表达。Western blot检测转染HS-746T细胞间质表皮转化因子(c-Met)/胆绿素还原酶(BVR)/活化复制因子2(ATF-2)分子通路相关蛋白的表达。结果:与GES-1细胞比较,HS-746T、BGC823、NCI-N87、SGC7901、AGS细胞中RP11-497E19.1表达水平均上升,且HS-746T细胞RP11-497E19.1表达水平最高(均P<0.05)。与si-NC组比较,si-RP11-497E19.1组HS-746T细胞活力和细胞侵袭数降低(均P<0.05)。双荧光素酶报告基因实验证实RP11-497E19.1靶向结合miR-545-5p,可负向调控miR-545-5p表达(P<0.05)。与si-NC组比较,si-RP11-497E19.1组HS-746T细胞c-Met/BVR/ATF-2分子通路蛋白c-Met、BVR、p-ATF-2、锌指蛋白1(Snail1)、锌指蛋白2(Snail2)表达降低(均P<0.05)。结论:胃癌细胞中RP11-497E19.1呈高表达,沉默RP11-497E19.1通过靶向miR-545-5p/c-Met/BVR/ATF-2分子通路抑制胃癌HS-746T细胞的增殖及侵袭。

Objective:To investigate the effect of lncRNA RP11-497E19.1 on the proliferation and invasion of gastric cancer cells and its mechanism.Methods:RT-qPCR was used to detect the expression of RP11-497E19.1 in gastric cancer cell lines HS-746T,BGC823,NCI-N87,SGC7901,AGS and immortalized gastric epithelial cell line GES-1,and the cells with the highest expression were selected for subsequent experiments.HS-746T cells were divided into si-NC group and si-RP11-497E19.1 group,which were transfected with negative control oligonucleotide or RP11-497E19.1 small interfering RNA,respectively.The proliferation and invasion ability were analyzed by colony formation assay and Transwell assay.The targeting relationship between RP11-497E19.1 and miR-545-5p was verified by dual luciferase reporter gene assay.The expression of miR-545-5p was detected by RT-qPCR.The expression of c-Met/BVR/ATF-2 pathway related proteins were detected by Western blot.Results:Compared with GES-1 cells,the expression levels of RP11-497E19.1 in HS-746T,BGC823,NCI-N87,SGC7901 and AGS cells were increased,and the expression level of RP11-497E19.1 in HS-746T cells was the highest(all P<0.05).Compared with the si-NC group,the proliferation and invasive number of HS-746T cells in the si-RP11-497E19.1 group were decreased(all P<0.05).Dual luciferase reporter gene assay confirmed that RP11-497E19.1 targeted miR-545-5p and negatively regulated the expression of miR-545-5p(P<0.05).Compared with the si-NC group,the expression of c-Met,BVR,p-ATF-2,Snail1 and Snail2 proteins in the c-Met/BVR/ATF-2 molecular pathway in HS-746T cells in the si-RP11-497E19.1 group decreased(all P<0.05).Conclusion:RP11-497E19.1 is highly expressed in gastric cancer cells.Silencing RP11-497E19.1 inhibits the proliferation and invasion of gastric cancer HS-746T cells by targeting the miR-545-5p/c-Met/BVR/ATF-2 molecular pathway.