miR-203和ZEB2直接相互作用逆转胃癌细胞顺铂耐药性的作用研究  

Direct interaction between miR-203 and ZEB2 reversed cisplatin resistance in gastric cancer

作　　者：王彩玲[1] 王俊生[1] 候新芳 周静[1] 阚随随 WANG Cai-ling;WANG Jun-sheng;HOU Xin-fang;ZHOU Jing;KAN Sui-sui(Department of Medicine,Anyang Tumor Hospital,Anyang 455000,China;Department of Oncology,Tumor Hospital Affiliated to Zhengzhou University,Henan Cancer Hospital,Zhengzhou 450008,China)

机构地区：[1]安阳市肿瘤医院内六科,河南安阳455000 [2]郑州大学附属肿瘤医院河南省肿瘤医院肿瘤内科,郑州450008

出　　处：《实用药物与临床》2019年第3期264-270,共7页Practical Pharmacy and Clinical Remedies

摘　　要：目的探讨miR-203和E盒结合锌指蛋白(ZEB2)直接相互作用对胃癌顺铂(DDP)耐药细胞株SGC-7901/DDP增殖和凋亡活性的影响。方法体外培养SGC-7901细胞和SGC-7901/DDP细胞,将miR-203 mimics或siRNA ZEB2转染至SGC-7901/DDP细胞,分别为miR-203组和Si-ZEB2组。另外,沉默SGC-7901/DDP细胞ZEB2基因的表达,采用MTT法和Annexin V/PI双染法检测SGC-7901细胞和SGC-7901/DDP细胞增殖和凋亡活力的改变。采用双荧光素酶报告基因方法验证miR-203与ZEB2的靶向关系。采用Western blot法检测各组细胞中上皮细胞-间充质转化(EMT)相关蛋白E-cadherin和Snail的表达。结果经MTT法检测,DDP对SGC-7901细胞和SGC-7901/DDP细胞的IC_(50)分别为18.37μmol/L和94.68μmol/L。SGC-7901/DDP的耐药指数为5.15±0.27。SGC-7901/DDP细胞中miR-203表达量低于SGC-7901细胞(P<0.05)。miR-203组细胞miR-203表达量较其他组明显升高(P<0.05)。另外,Si-ZEB2组细胞ZEB2 mRNA表达量降低,而miR-203表达量明显升高(P<0.05)。相同浓度条件下,DDP对miR-203组SGC-7901/DDP细胞增殖活性的抑制率明显高于SGC-7901/DDP组和NC组(P<0.05),同时,DDP对Si-ZEB2组细胞增殖活性的抑制率明显高于SGC-7901/DDP组和Si-NC组(P<0.05)。DDP(25μmol/L)对miR-203组细胞凋亡的促进作用明显高于SGC-7901/DDP组和NC组(P<0.05)。miR-203组细胞Snail蛋白表达量低于SGC-7901/DDP组和NC组,而E-cadherin蛋白表达量高于SGC-7901/DDP组和NC组(P<0.05)。另外,沉默SGC-7901/DDP细胞ZEB2基因表达后,E-cadherin蛋白表达量升高,而Snail蛋白的表达量降低(P<0.05)。结论 miR-203与ZEB2形成双向负反馈调节通路,通过上调E-cadherin的表达,抑制Snail的表达,阻止EMT进程,从而逆转胃癌细胞对顺铂的耐药性。Objective To discuss the direct interaction between miR-203 and zinc-finger E-box-binding protein 2(ZEB2)and their effects on reversing cisplatin resistance of SGC-7901/DDP cisplatin(DDP)resistant cells in gastric cancer.Methods SGC-7901 cells and SGC-7901/DDP cells were cultured in vitro.SGC-7901/DDP cells were transfected by miR-203 mimics(miR-203 group)or siRNA ZEB2(Si-ZEB2 group).In addition,the expression of ZEB2 gene in SGC-7901/DDP cells was silenced.The proliferations of SGC-7901 cells and SGC-7901/DDP cells were detected by MTT assay.The apoptosis of SGC-7901/DDP cells was detected by Annexin V/PE staining.The relative miR-203 and ZEB2 mRNA levels of SGC-7901/DDP cells were detected by qRT-PCR.The epithelial cadherin(E-cadherin)protein and Snail protein were detected by Western blot.Results MTT assay showed that the IC50 of DDP on SGC-7901 cells and SGC-7901/DDP cells were 18.37 μmol/L and 94.68 μmol/L.The resistance index of SGC-7901/DDP was 5.15±0.27.The relative miR-203 level of SGC-7901/DDP cells were lower than that of SGC-7901 cells(P<0.05),and miR-203 levels of SGC-7901/DDP cells in miR-203 group were higher than those in the other groups(P<0.05).There were lower ZEB2 level and higher miR-203 level in Si-ZEB2 group than that in Si-NC group(P<0.05).At the same concentration,the inhibition rate and apoptosis rate of DDP on SGC-7901/DDP cells in miR-203 group were higher than SGC-7901/DDP group and NC group(P<0.05);the inhibition rates and apoptosis rate of DDP in Si-ZEB2 group were higher than SGC-7901/DDP group and Si-NC group(P<0.05).The apoptotic effect of DDP(25 μmol/L)on the apoptosis of miR-203 cells was higher than that of SGC-7901/DDP group and NC group(P<0.05).There were lower Snail protein level and higher E-cadherin protein level of SGC-7901/DDP cells in miR-203 group than SGC-7901/DDP group and NC group(P<0.05).Besides,there were lower Snail protein level and higher E-cadherin protein level of SGC-7901/DDP cells in Si-ZEB2 group than SGC-7901/DDP group and Si-NC group(P<0.05).Conclus

关 键 词：miR-203 ZEB2 胃癌 顺铂 耐药性 

分 类 号：R735.2[医药卫生—肿瘤]