[1]陈青松,万磊,周壮,等.GSK-J4通过抑制STAT3磷酸化对HepG2肝癌细胞凋亡和侵袭的影响[J].第三军医大学学报,2018,40(09):739-745.
 CHEN Qingsong,WAN Lei,ZHOU Zhuang,et al.Jmjd3 inhibitor GSK-J4 induces apoptosis and attenuates invasion in HepG2 cells by inhibiting STAT3 phosphorylation [J].J Third Mil Med Univ,2018,40(09):739-745.
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GSK-J4通过抑制STAT3磷酸化对HepG2肝癌细胞凋亡和侵袭的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
40卷
期数:
2018年第09期
页码:
739-745
栏目:
基础医学
出版日期:
2018-05-15

文章信息/Info

Title:
Jmjd3 inhibitor GSK-J4 induces apoptosis and attenuates invasion in HepG2 cells by inhibiting STAT3 phosphorylation
 
作者:
陈青松万磊周壮周翔宇郑道峰吴忠均
重庆医科大学附属第一医院肝胆外科
Author(s):
CHEN Qingsong WAN Lei ZHOU Zhuang ZHOU Xiangyu ZHENG Daofeng WU Zhongjun

Department of Hepatobiliary Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
GSK-J4含Jumonji结构域蛋白3HepG2凋亡侵袭STAT3
Keywords:
GSK-J4 Jumonji domain-containing protein 3 HepG2 cells apoptosis invasion STAT3
分类号:
R73-361; R735.7; R979.19
文献标志码:
A
摘要:

目的    探讨GSK-J4通过抑制组蛋白去甲基化酶含Jumonji结构域蛋白3 (Jumonji domain-containing protein 3,Jmjd3)并上调H3K27me3的表达影响HepG2肝癌细胞凋亡和侵袭的分子机制。 方法    体外培养人正常肝细胞L02及人肝细胞肝癌(hepatocellular carcinoma,HCC)细胞株HepG2、SMMC7721,RT-PCR检测Jmjd3 mRNA表达,Western blot检测Jmjd3、H3K27me3蛋白表达;CCK-8法检测不同浓度GSK-J4 (0、10、30、50 μmol/mL)处理HepG2后增殖能力;流式细胞仪检测凋亡率;Transwell检测细胞侵袭力;Western blot检测Jmjd3、H3K27me3和凋亡相关蛋白Bcl2、Bax、Caspase3、上皮细胞间质转化(epithelial-mesenchymal transition,EMT)相关标记物E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)以及p-STAT3、STAT3蛋白表达。 结果    与L02比,Jmjd3高表达,H3K27me3低表达于HepG2、SMMC7721 肝癌细胞株(P<0.05);GSK-J4抑制HepG2增殖(P<0.05);GSK-J4处理组细胞凋亡率明显提高,细胞侵袭能力减弱(P<0.05);GSK-J4处理HepG2后Jmjd3水平降低,H3K27me3水平增高,Bcl2水平降低,Bax、Caspase3水平增高,E-cadherin表达增高,Vimentin水平降低(P<0.05);p-STAT3表达下调(P<0.01)。结论    GSK-J4通过抑制Jmjd3并上调H3K27me3表达而诱导HepG2发生凋亡并减弱侵袭,其可能与抑制EMT形成和STAT3的磷酸化有关。

Abstract:

Objective     To determine the effect of GSK-J4 on apoptosis and invasion in HepG2 cells by inhibiting histone demethylase Jumonji domain-containing protein 3 (Jmjd3) and modifying H3K27me3, and mine its underlying mechanism. Methods    Human normal hepatocytes L02 and hepatocellular carcinoma (HCC) cell lines HepG2 and SMMC7721 were employed in this study. The mRNA expression of Jmjd3 was detected by RT-PCR, and its protein level and that of H3K27me3 were measured by Western blotting. CCK-8 assay was used to detect the proliferation of HepG2 cells after the treatment of different concentrations of GSK-J4 (0, 10, 30 and 50 μmol/mL) treatment. Flow cytometry and Transwell chamber test were used respectively to detect cell apoptosis and invasion ability. The expression levels of Jmjd3 and H3K27me3, apoptosis related proteins Bcl2, Bax and Caspase3, epithelial-mesenchymal transition (EMT) markers E-cadherin and Vimentin, and p-STAT3 and STAT3 were detected by Western blotting. ResultsJmjd3 was significantly highly expressed, while H3K27me3 was lowly expressed in HCC cell lines HepG2 and SMMC7721 when compared with the L02 cells (P<0.05). GSK-J4 treatment inhibited the proliferation of HepG2 cells (P<0.05), increased cell apoptotic rate, weakened invasion ability (P<0.05), decreased the expression of Jmjd3 while elevated that of H3K27me3. The treatment also resulted in the decrease of Bcl2 level, increases of Bax and Caspase3 levels, elevation of E-cadherin, but reduce of Vimentin level (P<0.05), and inhibition of STAT3 phosphorylation (P<0.01). Conclusion     GSK-J4 induces apoptosis and suppresses the invasion ability in HepG2 cells by inhibiting Jmjd3 and up-regulating H3K27me3, which may be related to the suppression of EMT formation and STAT3 phosphorylation.

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更新日期/Last Update: 2018-05-10