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瞬时受体电位阳离子通道亚家族C成员6对子宫内膜癌细胞周期的调控作用
王朕华1*,李金凤1,杨海荣2,王悦1,井佳雨1
0
(1. 郑州大学人民医院、河南省人民医院妇科, 郑州 450003;
2. 焦煤集团中央医院妇科, 焦作 454150
*通信作者)
摘要:
目的 探讨子宫内膜癌组织中瞬时受体电位阳离子通道亚家族C成员6(TRPC6)的表达变化及其对子宫内膜癌细胞周期的调控作用。方法 应用qRT-PCR技术和蛋白质印迹法检测30例正常子宫内膜、30例不典型增生和32例子宫内膜癌组织中TRPC6的表达。分别用SKF59636(TRPC6阻断剂)和小干扰RNA(siRNA)干扰两种方法阻断TRPC6的作用,观察子宫内膜癌细胞HEC-1A细胞周期的变化。结果 子宫内膜癌组织中的TRPC6 mRNA和蛋白表达量均高于不典型增生组织和正常子宫内膜组织(P<0.01)。SKF96365呈剂量依赖性方式将细胞阻滞于G2/M期,使G0/G1期细胞减少;转染siRNA可将子宫内膜癌细胞阻滞于G2/M期,使G0/G1期细胞减少,同时上调磷酸化细胞分裂周期蛋白2(pCDC2)的表达。结论 子宫内膜癌组织中TRPC6表达增高,TRPC6可能通过影响pCDC2蛋白表达参与子宫内膜癌细胞周期调控。
关键词:  子宫内膜肿瘤  瞬时受体电位阳离子通道亚家族C成员6  小分子干扰RNA  细胞周期
DOI:10.16781/j.0258-879x.2020.01.0106
投稿时间:2019-07-12修订日期:2019-10-20
基金项目:河南省科技厅科技攻关项目(162102310022),河南省高等学校重点科研项目(13A320639),河南省医学科技攻关计划项目(201203120).
Transient receptor potential cation channel subfamily C member 6 is involved in regulating cell cycle of endometrial carcinoma cells
WANG Zhen-hua1*,LI Jin-feng1,YANG Hai-rong2,WANG Yue1,JING Jia-yu1
(1. Department of Gynecology, People's Hospital of Zhengzhou University, Henan Provicial People's Hospital, Zhengzhou 450003, Henan, China;
2. Department of Gynecology, Central Hospital of Jiaozuo Coalfield Group Corporation, Jiaozuo 454150, Henan, China
*Corresponding author)
Abstract:
Objective To study the expression of transient receptor potential cation channel subfamily C member 6 (TRPC6) in endometrial carcinoma tissues and their role in regulating cell cycle of endometrial carcinoma cells. Methods Quantitative real-time PCR and Western blotting were used to examine the expression of TRPC6 in 30 normal endometrial specimens, 30 atypical hyperplasia specimens and 32 endometrial carcinoma specimens. SKF96365 (an inhibitor of TRPC6 channel) and RNA interference (RNAi) targeting TRPC6 by small interference RNA (siRNA) were used to block TRPC6 so as to explore the role of TRPC6 in regulating the cell cycle of endometrial carcinoma cells HEC-1A. Results The expression levels of TRPC6 mRNA and protein in endometrial carcinoma were significantly higher than those in the atypical hyperplasia endometria and normal endometrial tissues (P<0.01). SKF96365 retarded cell cycle at G2/M phase in a dose-dependent manner and reduced HEC-1A cells of G0/G1 phase. Transfection with target-TRPC6 siRNA retarded cell cycle of HEC-1A cells at G2/M phase, and reduced HEC-1A cells of G0/G1 phase compared with negative control siRNA. Meanwhile, transfection with target-TRPC6 siRNA increased phosphorylated cell division cycle 2 (pCDC2) protein expression in HEC-1A cells. Conclusion The expression of TRPC6 is elevated in endometrial carcinoma tissues. TRPC6 may influence cell cycle through regulating pCDC2.
Key words:  endometrial neoplasms  transient receptor potential cation channel subfamily C member 6  small interfering RNA  cell cycle