[1]王 慧),耿 玲),郭 威),等.上调miR-15a表达对子宫内膜癌细胞增殖、凋亡和NOB1蛋白表达的影响[J].郑州大学学报(医学版),2019,(06):840-844.[doi:10.13705/j.issn.1671-6825.2019.06.031]
 WANG Hui),GENG Ling),GUO Wei),et al.Effects of up-regulating miR-15a on proliferation, apoptosis and NOB1 expression of endometrial carcinoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2019,(06):840-844.[doi:10.13705/j.issn.1671-6825.2019.06.031]
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上调miR-15a表达对子宫内膜癌细胞增殖、凋亡和NOB1蛋白表达的影响()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2019年06期
页码:
840-844
栏目:
应用研究
出版日期:
2019-11-20

文章信息/Info

Title:
Effects of up-regulating miR-15a on proliferation, apoptosis and NOB1 expression of endometrial carcinoma cells
作者:
王 慧1)耿 玲1) 郭 威1) 刘慧源1) 耿旭景2)
1)驻马店市中心医院生殖医学科 河南驻马店 463000 2)郑州大学第二附属医院生殖医学部 郑州 450014
Author(s):
WANG Hui1)GENG Ling1)GUO Wei1) LIU Huiyuan1)GENG Xujing2)
1)Department of Reproductive Medicine,Zhumadian Central Hospital,Zhumadian,Henan 463000 2)Department of Reproductive Medicine,the Second Affiliated Hospital,Zhengzhou University,Zhengzhou 450014
关键词:
子宫内膜癌 miR-15a NOB1 凋亡 增殖
Keywords:
endometrial cancer miR-15a NOB1 apoptosis proliferation
分类号:
R737.33
DOI:
10.13705/j.issn.1671-6825.2019.06.031
摘要:
目的:研究上调miR-15a表达对子宫内膜癌细胞增殖、凋亡和NOB1蛋白表达的影响。方法:将子宫内膜癌RL-952细胞分为3组,分别给予不转染(空白对照组)、转染miR-15a mimics(miR-15a组)或对照序列(miR-NC组)处理。转染48 h后,qRT-PCR法检测3组细胞中miR-15a表达水平,CCK-8检测细胞增殖能力,Western blot法检测细胞中Bax和Bcl-2蛋白的表达; 平板克隆实验测定细胞克隆形成能力; Annexin V-FITC/PI双染法检测凋亡。利用Targetscan预测NOB1的3'UTR端与miR-15a有结合位点,利用双荧光素酶报告实验鉴定两者的靶向关系。RL-952细胞共转染NOB1过表达载体和miR-15a mimics 48 h后,检测细胞增殖能力、克隆形成能力、凋亡率,以及细胞中Bax、Bcl-2、NOB1蛋白的表达。结果:与空白对照组、miR-NC组比较,miR-15a组细胞中miR-15a表达水平升高,细胞增殖能力和克隆形成率均降低,凋亡率升高,细胞中Bax表达水平升高,Bcl-2表达水平降低(P<0.05)。双荧光素酶报告实验证实miR-15a靶向负调控NOB1的表达。与转染miR-15a mimics的细胞比较,共转染NOB1过表达载体和miR-15a mimics的RL-952细胞增殖能力、克隆形成率升高,Bcl-2、NOB1蛋白表达升高,凋亡率和Bax表达降低(P<0.05)。结论:上调miR-15a表达可能通过靶向下调NOB1,抑制子宫内膜癌细胞的增殖,诱导癌细胞凋亡。
Abstract:
Aim:To study the effects of up-regulating miR-15a on proliferation, apoptosis and NOB1 expression of endometrial carcinoma cells.Methods:Endometrial cancer RL-952 cells were transfected with miR-15a mimics(miR-15a group)or the control sequence(miR-NC group), or without transfection(blank control group).After 48 hours,the miR-15a expression was detected by qRT-PCR, CCK-8 method was used to detect cell proliferation, Western blot was used to detect the expressions of Bax and Bcl-2; cloning capacity was determined by plate cloning assay; apoptosis was detected by Annexin V-FITC/PI staining. Targetscan predicted that the target gene of miR-15a might be NOB1,and double luciferase reporting experiment was used to identify the targeting relationship. RL-952 cells was co-transfected with NOB1 overexpression vector and miR-15a mimics,and 48 hours later the proliferation, cloning capacity,apoptosis,and the expressions of Bax,Bcl-2,NOB1 were detected.Results:Compared with those of the blank control group and miR-NC group, miR-15a expression, apoptosis rate,and Bax protein expression increased in miR-15a group, proliferation capacity, cloning capacity and Bcl-2 protein expression decreased(P<0.05). Double luciferase reporting experiment identified that miR-15a negatively regulate NOB1 protein expression. Compared with the RL-952 cells only transfected with miR-15a mimics,the proliferation, cloning capacity and the expressions of Bcl-2,NOB1 proteins in RL-952 cells co-transfected with NOB1 overexpression vector and miR-15a mimics increased,and apoptosis and Bax expression decreased(P<0.05).Conclusion:Up-regulating miR-15a expression may attenuate the proliferation of endometrial cancer cells and induce apoptosis by down-regulating NOB1.

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备注/Memo

备注/Memo:
【基金项目】河南省医学科技攻关计划项目(201802055) 【作者简介】耿旭景,通信作者,女,1987年7月生,博士生,主治医师,研究方向:生殖内分泌,E-mail:xj_geng@126.com
更新日期/Last Update: 2019-11-20