[1]熊浩君,赵凯,何凤田,等.5-氮杂-2′-脱氧胞苷抑制肾癌细胞增殖的机制研究[J].第三军医大学学报,2014,36(09):847-851.
 Xiong Haojun,Zhao Kai,He Fengtian,et al.5-aza-2′-deoxycytidine inhibits proliferation in renal carcinoma cells via suppressing methyltransferases and enhancing P21[J].J Third Mil Med Univ,2014,36(09):847-851.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
36卷
期数:
2014年第09期
页码:
847-851
栏目:
论著
出版日期:
2014-05-15

文章信息/Info

Title:
5-aza-2′-deoxycytidine inhibits proliferation in renal carcinoma cells via suppressing methyltransferases and enhancing P21
作者:
熊浩君赵凯何凤田曾益军陈姗陈丙波
第三军医大学:实验动物中心,基础医学部生物化学与分子生物学教研室
Author(s):
Xiong Haojun Zhao Kai He Fengtian Zeng Yijun Chen Shan Chen Bingbo
Laboratory Animal Center, Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Third Military Medical University, Chongqing, 400038, China
关键词:
5-氮杂-2′-脱氧胞苷肾细胞癌细胞增殖细胞信号抑制因子3P21
Keywords:
5-aza-2′-deoxycytidine renal carcinoma cells proliferationsuppressor of cytokine signaling 3 P21
分类号:
R730.23; R737.11; R979.19
文献标志码:
A
摘要:
目的      探讨5-氮杂-2′-脱氧胞苷(5-Aza-CdR)抑制肾癌细胞增殖的分子机制。      方法      采用不同浓度的5-Aza-CdR作用于769-P和ACHN 2种肾癌细胞,处理72 h后CCK-8法检测其对细胞增殖的影响;5-Aza-CdR处理769-P细胞48 h后,流式细胞仪测定细胞周期分布,RT-PCR测定甲基化转移酶DNMT1、DNMT3a、DNMT3b以及细胞信号抑制因子3(suppressor of cytokine signaling 3,SOCS3)的表达,Western blot测定SOCS3、P21在蛋白质水平的表达;siRNA干扰SOCS3表达以后,加5-Aza-CdR处理769-P细胞48 h,Western blot检测细胞内SOCS3、P21的表达。      结果      5-Aza-CdR可以剂量性依赖地抑制肾癌细胞系769-P和ACHN的增殖(P<0.05),且使769-P细胞阻滞在G2/M期。RT-PCR检测发现,5-Aza-CdR可以下调DNMT1和DNMT3a的表达和上调SOCS3的表达(P<0.05)。Western blot检测发现5-Aza-CdR可以促进SOCS3蛋白质的表达,并且伴随着P21蛋白的升高。在干扰SOCS3的表达后,用5-Aza-CdR处理769-P细胞48 h,P21的表达明显降低。       结论      5-Aza-CdR通过抑制甲基化转移酶,增强SOCS3介导P21的表达,从而抑制肾癌细胞769-P增殖。
Abstract:
Objective         To investigate the underlying molecular mechanism of 5-aza-2′-deoxycytidine (5-Aza-CdR) in inhibition of renal carcinoma cell proliferation.       Methods         Cell proliferation was assessed by CCK-8 assay after the renal carcinoma cell lines 769-P and ACHN were treated with 5-Aza-CdR at different concentrations for 72 h. Flow cytometry was used to detect cell cycle in 769-P cells after treatment of 5-Aza-CdR at different concentrations for 48 h. RT-PCR was used to detect the expression of methyltransferases DNMT1/3a/3b and suppressor of cytokine signaling 3 (SOCS3). The expression of P21 and SOCS3 at protein level was detected by Western blot analysis, and detected again in the 769-P cells with SOCS3 knockdown by small interfering RNA (siRNA) after 48 hours’ treatment of 5-Aza-CdR at different concentrations.       Results         CCK-8 assay revealed that 5-Aza-CdR remarkably inhibited the proliferation in the 2 cell lines in a dose-dependent manner (P<0.05). 5-Aza-CdR treatment also resulted in the 769-P cells arrested at G2/M phase. RT-PCR showed 5-Aza-CdR down-regulated DNMT1 and DNMT3a but up-regulated SOCS3 (P<0.05). Western blotting found that 5-Aza-CdR enhanced the expression of SOCS3 and P21. After knocking down the expression of SOCS3 of 769-P cells, we treated 769-P cells with 5-Aza-CdR for 48 h and found that  the up-regulation of P21 was weakened compared with control group.       Conclusion       5-Aza-CdR suppresses DNA methyltransferases, and enhances the expression of P21 mediated by SOCS3, and then inhibits the proliferation in renal carcinoma 769-P cells.

参考文献/References:

熊浩君, 赵凯, 何凤田, 等. 5-氮杂-2′-脱氧胞苷抑制肾癌细胞增殖的机制研究[J].第三军医大学学报,2014,36(9):847-851.

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更新日期/Last Update: 2014-05-05