|本期目录/Table of Contents|

miR-15b通过下调细胞因子诱导的杀伤细胞PD-1表达抑制宫颈癌细胞增殖

《现代肿瘤医学》[ISSN:1672-4992/CN:61-1415/R]

期数:
2019年20期
页码:
3587-3592
栏目:
论着(基础研究)
出版日期:
2019-09-08

文章信息/Info

Title:
miR-15b inhibits the proliferation of cervical cancer cells by down-regulating the expression of cytokine-induced killer cell PD-1
作者:
牛志军;?赵书君;?杨 立
郑州大学第三附属医院妇产科,河南 郑州 450000
Author(s):
Niu Zhijun;?Zhao Shujun;?Yang Li
Department of Obstetrics and Gynecology,the Third Affiliated Hospital of Zhengzhou University,Henan Zhengzhou 450000,China.
关键词:
宫颈癌;?微小RNA-15b;?程序性死亡蛋白-1;?细胞因子诱导的杀伤细胞;?细胞增殖
Keywords:
cervical cancer;?microRNA-15b;?programmed death protein-1;?cytokines-induced killer cell;?cell proliferation
分类号:
R737.33
DOI:
10.3969/j.issn.1672-4992.2019.20.009
文献标识码:
A
qq自动领红包软件:
目的:探讨微小RNA-15b(miR-15b)与程序性死亡蛋白-1(PD-1)在细胞因子诱导的杀伤细胞(CIK)抑制宫颈癌细胞增殖过程中的调控作用,并初步分析其中miR-15b与PD-1的可能作用机制。方法:采用传统方法培养CIK细胞,采用流式细胞仪检测CIK细胞免疫表型,CCK-8法检测CIK细胞对宫颈癌细胞(SiHa)增殖的影响。实时定量聚合酶链反应(qRT-PCR)及蛋白印迹(WB)法检测CIK细胞共培养前后SiHa细胞中miR-15b及PD-1表达水平。实验将CIK细胞共培养SiHa细胞分为3组:空白对照组(未进行任何处理),NC组(加入miR-15b阴性对照质粒),si-miR-15b组(加入miR-15b siRNA)。采用qRT-PCR与WB分别检测各组miR-15b及PD-1表达水平;CCK-8法检测各组SiHa细胞增殖情况。采用WB法检测细胞增殖相关蛋白增殖细胞核抗原(PCNA)及Ki67表达。结果:CIK细胞共培养组SiHa细胞增殖抑制率显着高于单纯培养SiHa细胞组(P<0.05);CIK细胞共培养后SiHa细胞中miR-15b表达水平显着高于单纯培养SiHa细胞组(P<0.05),而PD-1蛋白表达水平显着降低(P<0.05);si-miR-15b组miR-15b表达水平、SiHa细胞增殖抑制率均显着低于空白对照组、NC组(P<0.05),而PD-1、PCNA、Ki67蛋白表达均显着升高(P<0.05)。结论:miR-15b可能通过下调CIK细胞上PD-1表达进而抑制宫颈癌细胞增殖。
Abstract:
Objective:To investigate the regulatory role of microRNA-15b (miR-15b) and programmed death protein-1 (PD-1) in cytokine-induced killer cells (CIK) inhibiting the proliferation of cervical cancer cells,and to preliminarily analyze the possible mechanism of miR-15b and PD-1.Methods:CIK cells were cultured by traditional methods.The immunophenotype of CIK cells was detected by flow cytometry.The effect of CIK cells on the proliferation of cervical cancer cells (SiHa) was detected by CCK-8 method.Real-time quantitative polymerase chain reaction (qRT-PCR) and Western blot (WB) were used to detect the expressions of miR-15b and PD-1 in SiHa cells before and after co-culture of CIK cells.SiHa cells co-cultured with CIK cells were divided into three groups:Blank control group (without any treatment),NC group (with negative control plasmid of miR-15b) and si-miR-15b group (with miR-15b siRNA).The expression levels of miR-15b and PD-1 in each group were detected by qRT-PCR and WB,respectively.CCK-8 method was used to detect the proliferation of SiHa cells in each group.WB method was used to detect the expressions of proliferating cell nuclear antigen (PCNA) and Ki67.Results:The proliferation inhibition rate of SiHa cells in CIK cells co-culture group was significantly higher than that in SiHa cells alone-culture group (P<0.05).After co-culture of CIK cells,the expression of miR-15b in SiHa cells was significantly higher than that in SiHa cells alone-culture group (P<0.05).While the expression of PD-1 protein was significantly lower (P<0.05).The expression level of miR-15b and the inhibition rate of proliferation of SiHa cells in si-miR-15b group were significantly lower than those in blank control group and NC group (P<0.05),while the expressions of PD-1,PCNA,and Ki67 protein were significantly increased (P<0.05).Conclusion:miR-15b may inhibit the proliferation of cervical cancer cells by down-regulating the expression of PD-1 on CIK cells.

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备注/Memo

备注/Memo:
河南省高等学校重点科研项目(编号:19A320052)
更新日期/Last Update: 1900-01-01