长链非编码RNA MEG3靶向调控Rb1抑制上皮性卵巢癌细胞增殖研究  

Long non-coding RNA MEG3 inhibit epithelial ovarian cancer cells proliferation via targeting rb1 pathway

作　　者：李剑琦[1] 刘艳 周冬梅[1] 汪志辉[1] 生秀杰[1] LI Jian-qi;LIU Yan;ZHOU Dong-mei;WANG Zhi-hui;SHENG Xiujie(Department of Obstetrics and Gynecology,Third Affiliated Hospital of Guangzhou Medical University,Key Laboratory of Major Obstetric Diseases of Guangdong Province,Key Laboratory of Reproduction and Genetic of Guangdong Higher Education Institutes,Guangzhou 510150,China)

机构地区：[1]广州医科大学附属第三医院妇科,广东省产科重大疾病重点实验室,广东省普通高校生殖与遗传重点实验室,广东广州510150

出　　处：《中华肿瘤防治杂志》2021年第20期1555-1561,共7页Chinese Journal of Cancer Prevention and Treatment

基　　金：广州市医药卫生科技项目(20161A011078)。

摘　　要：目的研究母系表达基因3(MEG3)对上皮性卵巢癌细胞埴殖影响及其作用机制。方法逆转录-聚合酶链反应(RT-PCR)检测67例人上皮性卵巢癌组织和20例正常卵巢组织中MEG3表达。实验设置MEG3过表达质粒组(MEG3组)和空质粒pcDNA3.1对照组(NC组)。过表达MEG3质粒转染卵巢癌细胞OVCAR3和SKOV3,MTT实验、细胞周期和细胞凋亡实验检测MEG3对卵巢癌细胞生长的影响;双荧光素酶报告系统检测MEG3靶基因Rb1的转录活性;RT-PCR和蛋白质印迹法检测Rb1 mRNA和蛋白表达水平。结果正常卵巢组织中MEG3表达量为3.32±0.25,上皮性卵巢癌组织为0.35±0.11,差异有统计学意义,t=10.932,P<0.001.转染MEG3过表达质粒后,12、24、36和48 h的OVCAR3和SKOV3细胞生长抑制率分别为(37.26±2.21)%和(23.06±1.67)%、(38.19±1.98)%和(42.71±2.36)%、(44.30±2.14)%和(44.31±3.38)%、(47.67±4.32)%和(50.36±2.76)%,差异有统计学意义,F值分别为131.203和353.782,均P<0.001;转染MEG3过表达质粒后OVCAR3和SKOV3细胞S期比例下降.差异有统计学意义(t=7.471,P<0.001;t=5.197,P<0.001);OVCAR3和SKOV3细胞凋亡率增加,差异有统计学意义(t=6.820,P=0.029,t=4.235,P<0.001)。对比NC组.MEG3组Rb1转录活性增加,OVCAR3和SKOV3细胞RB1 mRNA表达分别增加(1.74±0.54)和(1.95±0.72)倍,差异有统计学意义(t=24.504,P<0.001,t=28.276.P<0.001);OVCAR3和SKOV3细胞Rb1蛋白表达分别增加(1.92±0.32)和(1.75±0.25)倍,差异有统计学意义(t=10.500.P<0.001,t=9.375,P<0.001)。结论MEG3在上皮性卵巢癌组织和细胞中表达降低,推测可能与上皮性卵巢癌的发生发展相关。MEG3可能通过调控Rb1抑制上皮性卵巢癌细胞增殖并促进凋亡。Objective To study the effect and mechanism of maternally expressed gene 3(MEG3)on epithelial ovarian cancer cell proliferation.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of MEG3 in 67 human epithelial ovarian cancer tissues and 20 normal ovarian tissues.The experiment was set up aMEG3 overexpression plasmid group(MEG3 group)and an empty plasmid pcDNA3.1 control group(NC group).Overexpression of MEG3 plasmid was transfected into ovarian cancer cells OVCAR3 and SKOV3.MTT experiments,cell cycle and apoptosis experiments were used to detect the effect of MEG3 on the growth of ovarian cancer cells;dual luciferase reporter system was detected the transcriptional activity of MEG3 target gene Rb1.RT-PCR and Western blotting were used to detect the mRNA and protein expression levels of Rb1.Results The expression of MEG3 in normal ovarian tissue was 3.32±0.25,and that in epithelial ovarian cancer tissue was 0.35±0.11,the difference was statistically significant(t=10.932,P<0.001).After transfection of MEG3 overexpression plasmid,the growth inhibition rates of OVCAR3 and SKOV3 cells at 12,24,36 and 48 hwere(37.26±2.21)% and(23.06±1.67)%,(38.19±1.98)% and(42.71±2.36)%,(44.30±2.14)% and(44.31±3.38)%,(47.67±4.32)% and(50.36±2.76)%,respectively,and the difference was statistically significant(Fvalues were 131.203 and 353.782,both P<0.001).The ratio of S stage declined after transfection of MEG3 overexpression plasmid,the difference was statistically significant(t=7.471,P<0.001;t=5.197,P<0.001).The apoptotic rate of OVCAR3 and SKOV3 cells increased,the difference was statistically significant(t=6.820,P=0.029;t=4.235,P<0.001).Compared with NC group,MEG3 group Rb1 transcription activity increased,the OVCAR3 and SKOV3 cells Rb1 mRNA expression increased by(1.74±0.54)and(1.95±0.72)times,respectively,and the difference was statistically significant(t=24.504,P<0.001;t=28.276,P<0.001).The the OVCAR3 and SKOV3 cells Rb1 protein expression increased by(1.92±0.32)times and

关 键 词：上皮性卵巢癌 母系表达基因3 长链非编码RNA 细胞增殖 

分 类 号：R737.31[医药卫生—肿瘤]