下调EZH2提高胃癌SGC7901细胞对奥沙利铂的敏感性

doi:10.3760/cma.j.issn.1673422X.2018.02.001

摘要/Abstract

摘要： 目的探讨下调zeste基因增强子同源物2（EZH2）基因对胃癌SGC7901细胞奥沙利铂敏感性的影响。方法针对EZH2 mRNA序列设计合成小干扰RNA（siRNA）片段，将其分为siRNA1组、siRNA2组、siRNA3组，转染至胃癌细胞株SGC7901中，同时设立阴性对照组和空白对照组，分别采用荧光定量PCR和Western blotting方法检测EZH2基因mRNA及其蛋白的表达情况，采用CCK8法检测不同浓度奥沙利铂处理各组胃癌细胞的增殖抑制率。结果转染EZH2 siRNA后，siRNA1组、siRNA2组、siRNA3组EZH2 mRNA的相对表达量分别为0.615±0.190、0.241±0.152、0.450±0.097，3组均低于阴性对照组(1.165±0.376)，且差异具有统计学意义(P=0.028；P=0.002；P=0.007)。SGC7901胃癌细胞转染最佳siRNA片段后，干扰组的EZH2蛋白相对表达量为0.036±0.017，空白对照组和阴性对照组分别为0.362±0.026、0.398±0.036，3组间差异有统计学意义(F=157.745，P＜0.001)，干扰组与阴性对照组、空白对照组相比，差异有统计学意义(P=0.001，P=0.002)。奥沙利铂浓度为2、4、8、16μg/ml时，干扰组、阴性对照组和空白对照组细胞增殖抑制率的差异有统计学意义［(18.107±2.822)%、(5.867±2.272)%、(5.333±1.883)%，F=28.185，P=0.001；(54.953±2.550)%、(22.177±1.871)%、(20.077±6.032)%，F=74.206，P＜0.001；(60.337±1.641)%、(34.597±3.592)%、(30.227±5.273)%，F=54.897，P＜0.001；(78.340±2.081)%、(61.857±3.507)%、(63.077±8.473)%，F=8.586，P=0.017］，而奥沙利铂浓度为32μg/ml时3组间差异无统计学意义［(83.450±3.715)%、(72.190±3.948)%、(70.731±17.080)%，F=1.358，P=0.326］。干扰组、阴性对照组及空白对照组的奥沙利铂半数抑制浓度（IC50）分别为5.178、12.643、13.601μg/ml。结论下调EZH2基因的表达能显著抑制胃癌细胞增殖，提高胃癌细胞对奥沙利铂的敏感性，说明EZH2基因在胃癌化疗进展过程中有很重要的意义，为深入研究胃癌基因治疗提供一定的理论依据。

关键词: 胃肿瘤, RNA, 小分子干扰, zeste基因增强子同源物2, 奥沙利铂

Abstract: ObjectiveTo investigate the effect of enhancer of zeste homolog 2 (EZH2) knockdown on chemosensitivity to oxaliplatin in human gastric cancer cell line SGC7901. MethodsSmall interfering RNA (siRNA) fragments were designed and synthesized for EZH2 mRNA sequence and divided into siRNA1 group, siRNA2 group and siRNA3 group. They were transfected into gastric cancer cell line SGC7901. The negative control group and blank control group were set up at the same time. The expressions of EZH2 mRNA and protein in the SGC7901 cells were detected by quantitative realtime polymerase chain reaction (qRTPCR) and Western blotting. CCK8 assay was used to detect cell proliferation inhibition rates of SGC7901 cells treated by different concentrations of oxaliplatin. ResultsAfter transfection of EZH2 siRNA, the relative expression levels of EZH2 mRNA in siRNA1 group, siRNA2 group, siRNA3 group were 0.615±0.190, 0.241±0.152 and 0.450±0.097. The relative expression levels of EZH2 mRNA of the three groups were lower than that in the negative control group (1.165±0.376), and the differences were statistically significant (P=0.028; P=0.002; P=0.007). After transfection of the best siRNA fragment into SGC7901 gastric cancer cells, the relative expression levels of EZH2 protein in interference group, blank control group and negative control group were 0.036±0.017, 0.362±0.026 and 0.398±0.036, and the difference among the three groups was statistically significant (F=157.745, P＜0.001). The difference between interference group and negative control group was statistically significant (P=0.001), as compared with the blank control group (P=0.002). When oxaliplatin concentration was 2, 4, 8 and 16μg/ml, the differences of cell proliferation inhibition rate among interference group, negative control group and blank control group were statistically significant ［(18.107±2.822)%, (5.867±2.272)%, (5.333±1.883)%,F=28.185, P=0.001; (54.953±2.550)%, (22.177±1.871)%, (20.077±6.032)%, F=74.206, P＜0.001; (60.337±1.641)%, (34.597±3.592)%, (30.227±5.273)%, F=54.897, P＜0.001; (78.340±2.081)%, (61.857±3.507)%, (63.077±8.473)%, F=8.586, P=0.017］. There was no significant difference among groups of oxaliplatin at the concentration of 32μg/ml ［(83.450±3.715)%, (72.190±3.948)%, (70.731±17.080)%, F=1.358, P=0.326］. The median inhibitory concentration (IC50) of oxaliplatin in siRNA group, negative control group and blank control group were 5.178, 12.643, 13.601μg/ml. ConclusionDownregulation of EZH2 gene expression can significantly inhibit the proliferation of gastric cancer cells, and effectively enhance the sensitivity of gastric cancer cells to oxaliplatin, which indicates EZH2 may play important roles in the development of gastric cancer chemotherapy. These results provide an important theoretical basis for gene therapy of gastric cancer.

Key words: Stomach neoplasms, RNA, small interfering, Enhancer of zeste homolog 2, Oxaliplatin

万倩, 李弦, 邓野, 王丹, 杨继元. 下调EZH2提高胃癌SGC7901细胞对奥沙利铂的敏感性[J]. 国际肿瘤学杂志, 2018, 45(2): 65-69.

WAN Qian, LI Xian, DENG Ye, WANG Dan, YANG Ji-Yuan. ncreasing oxaliplatin sensitivity by down-regulation the expression of EZH2 in human gastric cancer SGC7901 cells[J]. Journal of International Oncology, 2018, 45(2): 65-69.

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