沉默KDM5A基因对卵巢癌顺铂耐药SKOV3/DDP细胞迁移及侵袭的影响  

Silencing KDM5A gene inhibits the migration and invasion of ovarian cancer cisplatin-resistant SKOV3/DDP cells

作　　者：汪黎明[1] 冯同富[1] 陈竹 张纯[3] 黄燕明[1] 李力[2] WANG Li-ming;FENG Tong-fu;CHEN Zhu;ZHANG Chun;HUANG Yan-ming;LI Li(Maternal and Child Health Hospital of Hubei Province,Wuhan 430070,China;Key Laboratory of High Incidence Tumor Prevention and Treatment,Ministry of Education,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]湖北省妇幼保健院妇科,湖北武汉430070 [2]广西医科大学区域性高发肿瘤早期防治研究教育部重点实验室,广西南宁530021 [3]湖北省妇幼保健院生殖中心,湖北武汉430070

出　　处：《中华肿瘤防治杂志》2021年第12期903-908,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：湖北省自然科学基金知识创新专项(2018CFC840);湖北省卫计委联合基金(WJ2018H0158);武汉市中青年医学骨干人培养工程([2018]116)。

摘　　要：目的探讨沉默组蛋白赖氨酸去甲基化酶5A(KDM5A)基因表达对卵巢癌顺铂(DDP)耐药细胞SKOV3/DDP迁移及侵袭的影响。方法通过RNAi技术将细胞分为3组:转染沉默组(SKOV3/DDP-KDM5Ai)、阴性对照组(SKOV3/DDP-NC)和未转染的空白对照组(SKOV3/DDP),蛋白质印迹法验证KDM5A基因的沉默效果,划痕实验检测3组细胞的迁移能力,Transwell侵袭实验检测3组细胞的侵袭能力,逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹法检测3组细胞中β-catenin、糖原合酶激酶-3β(GSK-3β)和基质金属蛋白酶2(MMP2)的表达变化情况。采用单因素方差分析法比较各组间差异。结果转染沉默组、阴性对照组及空白组细胞中KDM5A蛋白的相对表达量分别为0.327 4±0.04、0.801 9±0.05和0.821 8±0.05,差异有统计学意义,F=131.466,P<0.001;其中,沉默组的表达强度低于阴性对照组(t=13.744,P<0.001)和空白组(t=14.323,P<0.001),差异有统计学意义。划痕实验结果显示,转染沉默组、阴性对照组及空白组细胞的划痕愈合率分别为(28.47±2.72)%、(50.58±4.06)%和(48.92±4.63)%,差异有统计学意义,F=30.158,P=0.007;其中,沉默组细胞的划痕愈合率低于阴性对照组(t=6.973,P<0.001)和空白组(t=6.447,P<0.001),差异有统计学意义。侵袭实验结果显示,转染沉默组、阴性对照组及空白组细胞的穿膜细胞数分别为(55.20±2.39)、(82.60±5.18)和(80.80±7.26)个,差异有统计学意义,F=41.354,P<0.001;其中,沉默组细胞的侵袭能力低于阴性对照组(t=8.129,P=0.005)和空白组(t=7.595,P=0.007),差异有统计学意义。RT-PCR和蛋白质印迹法检测结果显示,沉默组β-catenin、GSK-3β和MMP2的mRNA及蛋白表达水平低于阴性对照组和空白组,差异有统计学意义,P<0.01。结论沉默KDM5A基因可显著抑制SKOV3/DDP细胞的迁移及侵袭能力,这一过程可能与抑制Wnt/β-catenin信号通路有关。Objective To investigate the effect of silencing Lysine(K)demethylase 5 A(KDM5 A) gene expression on the migration and invasion of ovarian cancer cell line SKOV3/DDP. Methods The cells were divided into three groups by RNAi technique: silenced group(SKOV3/DDP-KDM5 Ai),negative control group(SKOV3/DDP-NC) and blank control group(SKOV3/DDP). Western blot method was used to verify the silencing effect of KDM5 A gene;wound healing assay was used to detect the migration ability of the cells;transwell invasion assay was used to detect the invasive ability of the cells;and RT-PCR and western blot were used to detected the changes of β-catenin,glycogen synthasc kinase-3β(GSK-3β) and matrix metalloproteinase 2(MMP2) in the three groups. Single factor analysis of variance was used to compare the differences among groups. Results The relative expression of KDM5 A protein in silenced group,negative control group and blank group was 0.327 4±0.04,0.801 9±0.05 and 0.821 8±0.05,respectively,and the difference was statistically significant(F=131.466,P<0.001).Among them,and the expression level in silenced group was significantly lower than that in negative control group and blank group(t=13.744,P<0.001;t=14.323,P<0.001).The wound healing assay results showed that the healing rates of silenced group,negative control group and blank group were(28.47±2.72)%,(50.58±4.06)% and(48.92±4.63)%,respectively,the difference was statistically significant(F=30.158,P=0.007).The healing rate in the silenced group was significantly lower than that in the negative control group and the blank group(t=6.973,P<0.001;t=6.447,P<0.001),the difference was statistically significant.The transwell invasion assay showed that the numbers of migrated cells in silenced group,negative control group and blank group were55.20±2.39,82.60±5.18 and 80.80±7.26 respectively,the difference was statistically significant(F=41.354,P<0.001),among which,the invasive ability of silent group was significantly lower than that of negative control group and blank grou

关 键 词：卵巢癌 肿瘤 组蛋白赖氨酸去甲基化酶5A 迁移 侵袭 WNT/Β-CATENIN信号通路 

分 类 号：R737.31[医药卫生—肿瘤]