miR-548c-5p靶向Notch通路增强胃癌SGC7901/VCR细胞对紫杉醇的敏感性  

MiR-548c-5p Targets Notch Pathway Enhances Gastric Cancer SGC7901/VCR Cells′ Susceptibility to Paclitaxel

作　　者：黄河 魏童[2] HUANG He;WEI Tong(Department of Gastroenterology,the Fifth Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China)

机构地区：[1]新疆医科大学第五附属医院胃肠外科,乌鲁木齐830054 [2]新疆医科大学第五附属医院血管肿瘤介入科,乌鲁木齐830054

出　　处：《实用医学杂志》2020年第15期2066-2071,共6页The Journal of Practical Medicine

基　　金：新疆维吾尔自治区自然科学基金资助项目(编号:2019D01C270)。

摘　　要：目的分析miR-548c-5p在胃癌SGC7901/VCR细胞药物耐受中的作用。方法通过qRTPCR分析miR-548c-5p在SGC7901细胞系和SGC7901耐药细胞系(SGC7901/VCR)中的表达差异。分别通过qRT-PCR和western blot分析SGC7901细胞系和SGC7901/VCR中Notch1和Hes1在mRNA水平上和蛋白质水平上的表达变化。通过荧光素酶实验分析miR-548c-5p与Notch1的3′-UTR的作用。采用CCK-8实验分析miR-548c-5p对细胞增殖的影响,采用Transwell实验分析miR-548c-5p对细胞侵袭的影响。在SGC7901/VCR细胞中转染miR-548c-5p mimics,采用CCK-8分析miR-548c-5p联合紫杉醇对细胞增殖的影响,采用流式细胞术分析miR-548c-5p联合紫杉醇对细胞增殖和侵袭的影响,评估miR-548c-5p对胃癌耐药细胞系SGC7901/VCR细胞药物耐受性的作用。结果 miR-548c-5p在SGC7901/VCR细胞中的表达显著低于SGC7901细胞(P <0.05),Notch1和Hes1 mRNA和蛋白质在SGC7901/VCR细胞中的表达显著低于SGC7901细胞(P <0.05)。miR-548c-5p mimics可以抑制SGC7901/VCR细胞的增殖和侵袭。而过表达Notch1可以抵消miR-548c-5p对SGC7901/VCR细胞的抑制作用。miR-548c-5p与紫杉醇联合使用,可以进一步抑制SGC7901/VCR细胞的增殖和侵袭。结论 miR-548c-5p可以通过靶向Notch通路,提高胃癌SGC7901/VCR细胞对化疗药物紫杉醇的敏感性。Objective To analyze the role of mir-548 c-5 p in drug tolerance of gastric cancer SGC7901/VCR cells.Methods The difference of miR-548 c-5 p expression in SGC7901 cell line and SGC7901 drug-resistant cell line(SGC7901/VCR) was analyzed by qRT-PCR.The expressions of Notchl and Hesl in SGC7901 cell line and SGC7901/VCR were analyzed by qRT-PCR and Western blot at mRNA level and protein level,respectively.The targeting of mir-548 c-5 p on the 3’-UTR of Notchl was analyzed by luciferase assay.CCK-8 experiment was used to analyze the effects of miR-548 c-5 p on cell proliferation,and Transwell experiment was used to analyze the effects of mir-548 c-5 p on cell migration and invasion.miR-548 c-5 p mimics were transfected into SGC7901/VCR cells.CCK-8 was used to analyze the effect of miR-548 c-5 p combined with paclitaxel on cell proliferation,flow cytometry was used to analyze the effect of miR-548 c-5 p combined with paclitaxel on cell apoptosis,and the role of mir-548 c-5 p in drug tolerance of gastric cancer resistant cell line SGC7901/VCR cells was evaluated.Results The expression of miR-548 c-5 p in SGC7901/VCR cells was significantly lower than that in SGC7901/VCR cells(P <0.05),and the mRNA and protein expressions of Notchl and Hesl in SGC7901/VCR cells were significantly lower than that in SGC7901/VCR cells(P <0.05).miR-548 c-5 p mimics could inhibit the proliferation and invasion of SGC7901/VCR cells.Overexpression of Notchl could counteract the inhibitory effect of mir-548 c-5 p on SGC7901/VCR cells.Combined with paclitaxel,mir-548 c-5 p further inhibited the proliferation of SGC7901/VCR cells.Conclusion miR-548 c-5 p can increase the sensitivity of gastric cancer SGC7901/VCR cells to the chemotherapy drug paclitaxel by targeting Notch pathway.

关 键 词：miR-548c-5p NOTCH通路 SGC7901/VCR细胞 化疗敏感性 

分 类 号：R735.2[医药卫生—肿瘤]