Twist-siRNA对SGC7901胃癌细胞上皮间质转化过程及顺铂耐药性的影响  

Effect of Twist-siRNA on epithelial-mesenchymal transition and cisplatin resistance in SGC7901 gastric cancer cells

作　　者：杨晓瑞 王江 张玮 刘凡凡 丁孝良 张凯 何勐 YANG Xiao-rui;WANG Jiang;ZHANG Wei;LIU Fan-fan;DING Xiao-liang;ZHANG Kai;HE Meng(Department of Pharmacy,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,Henan Province,China;Department of Gastrointestinal and Abdominal Surgery,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou 450000,Henan Province,China)

机构地区：[1]郑州大学附属郑州中心医院药学部,河南郑州450000 [2]郑州大学附属郑州中心医院胃肠疝和腹壁外科,河南郑州450000

出　　处：《中国临床药理学杂志》2020年第22期3666-3669,3678,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的观察应用小干扰RNA(siRNA)技术沉默上皮间质转化因子(Twist)对SGC7901胃癌细胞上皮间质转化(EMT)过程及顺铂耐药性的影响。方法以脂质体转染法将Twist-siRNA、对照siRNA转染至SGC7901细胞,设为siRNA组和N-siRNA组,对照组未给予任何处理,检测各组Twist mRNA和蛋白表达情况及细胞增殖、迁移、侵袭情况。测定不同浓度顺铂(6.25×10-2,0.125,0.25,0.50,1.00 mg·L^-1)对细胞的生长抑制率及顺铂对细胞的半数抑制浓度(IC50)。以0.50 mg·L^-1顺铂干预SGC7901细胞为顺铂组,以0.50 mg·L^-1顺铂干预稳定转染Twist-siRNA的SGC7901细胞为siRNA+顺铂组,检测各组上皮性钙黏附蛋白(E-cadherin)、波形蛋白(Vinentin)、磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(Akt)mRNA和蛋白及磷酸化AKT(pAKT)蛋白表达情况。结果 siRNA组Twist mRNA及蛋白相对表达量、MTT实验不同时刻OD值均低于对照组和N-siRNA组(均P<0.05)。siRNA组、对照组和N-siRNA组的迁移率分别为(68.11±6.42)%,(82.16±6.65)%和(85.50±6.25)%;穿膜细胞数分别为(46.67±4.64),(127.67±9.39)和(131.33±8.96)个;顺铂对细胞的IC50值分别为(0.19±0.01),(0.50±0.05)和(0.51±0.05)mg·L^-1,siRNA组均显著低于对照组和N-siRNA组(均P<0.05)。siRNA组、顺铂组、siRNA+顺铂组E-cadherin mRNA和蛋白相对表达量均高于对照组;siRNA组、顺铂组、siRNA+顺铂组Vinentin、PI3K mRNA和蛋白相对表达量及pAKT/AKT均低于对照组(均P<0.05);与siRNA组、顺铂组比较,siRNA+顺铂组E-cadherin mRNA和蛋白相对表达量均升高,Vinentin、PI3K mRNA和蛋白相对表达量及pAKT/AKT均降低(均P<0.05)。结论 Twist-siRNA可阻断SGC7901胃癌细胞EMT过程,降低细胞对顺铂的耐药性,可能与上调E-cadherin mRNA和蛋白表达,下调Vinentin、PI3K mRNA和蛋白表达,抑制AKT磷酸化相关。Objective To observe the effect of small interference RNA(siRNA) silencing on the process of epithelial mesenchymal transition(EMT) and cisplatin resistance in SGC7901 gastric cancer cells. Methods Twist siRNA and control siRNA were transfected into SGC7901 cells by liposome transfection method as siRNA group and N-siRNA group. Control group was given no treatment. The expression of Twist mRNA and protein and cell proliferation, cell migration and invasion in each group were compared. The inhibitory rates of different concentrations of cisplatin(6.25×10-2, 0.125, 0.25, 0.50, 1.00 mg·L^-1) on cell growth and the half inhibitory concentration(IC50) of cisplatin on cell growth were determined.SGC7901 cells treated with 0. 50 mg·L^-1 cisplatin were set as cisplatin group,and SGC7901 cells stably transfected with twist siRNA were treated with 0. 50 mg·L^-1 cisplatin as siRNA + cisplatin group. The expressions of E-cadherin,Vinentin,phosphatidylinositol-3 kinase(PI3 K) and protein kinase B(AKT) and phosphorylated AKT(pAKT) were detected. Results The relative expressions of Twist mRNA and protein,OD value at different time in MTT test were lower than those in control group and N-siRNA group(all P < 0. 05). The mobility rates in siRNA group,control group and N-siRNA group were(68. 11 ± 6. 42) %,(82. 16 ± 6. 65) % and(85. 50 ± 6. 25) %,the number of transmembrane cells were 46. 67 ± 4. 64,127. 67 ± 9. 39 and 131. 33 ± 8. 96,the IC50 values were(0. 19 ± 0. 01),(0. 50 ± 0. 05) and(0. 51 ± 0. 05) mg·L^-1,and the IC50 values of siRNA group were significantly lower than those in control group and N-siRNA group(all P < 0. 05). The relative expressions of E-cadherin mRNA and protein in siRNA group,cisplatin group,siRNA + cisplatin group were higher than those in control group and N-siRNA group(all P < 0. 05). The relative expressions of Vinentin,PI3 K mRNA and protein and p AKT/AKT in siRNA group,cisplatin group,siRNA + cisplatin group were lower than those in control group and N-siRNA group(all P < 0. 05). Compare

关 键 词：胃癌 小干扰RNA技术 上皮间质转化因子 上皮间质转化 顺铂耐药性 

分 类 号：R97[医药卫生—药品]