人脐带间充质干细胞通过IL-6/STAT3信号通路促进肝癌HepG-2细胞的增殖  

Human umbilical cord mesenchymal stem cells promote the proliferation of HepG-2 cells through interleukin-6/STAT3 signaling pathway

作　　者：郑盛[1] 杨涓[1] 陈文钦[2] 刘晶[2] 张帆[1] 王玉波[1] Zheng Sheng Yang Juan Chen Wen-qin Liu Jing Zhang Fan Wang Yu-bo(Department of Digestive Diseases, Third People＇s Hospital of Yunnan Province, Kunming 650011, Yunnan Province, China Department of Obstetrics, Dong Fang Hospital of Kunming, Kunming 650000, Yunnan Province, China)

机构地区：[1]云南省第三人民医院消化内科,云南省昆明市650011 [2]昆明市东方医院产科,云南省昆明市650000

出　　处：《中国组织工程研究》2016年第50期7460-7468,共9页Chinese Journal of Tissue Engineering Research

基　　金：云南省自然科学基金(2012FD095);云南省教育厅科研基金重点项目(2014Z125;2015Z146);云南省临床重点专科建设项目(云卫医发[2015]18号)~~

摘　　要：背景:人脐带间充质干细胞能分泌多种细胞因子,参与调控肿瘤的增殖、转移及血管生成等生物学行为,其中白细胞介素6可能是最重要的炎性因子之一。目的:探讨人脐带间充质干细胞通过IL-6/STAT3信号通路对肝癌HepG-2细胞增殖和迁移的作用。方法:采用ELISA法检测人脐带间充质干细胞和肝癌HepG-2细胞中白细胞介素6的表达量。Western blot法检测肝癌HepG-2细胞内STAT3以及p-STAT3蛋白质的表达水平。RT-PCR法检测PCNA、Cyclin D1、Survivin、STAT3基因的转录水平。细胞计数法和CCK-8法检测肝癌HepG-2细胞的增殖能力。划痕实验和Transwell实验检测肝癌HepG-2细胞的迁移能力。结果与结论:(1)人脐带间充质干细胞白细胞介素6表达量明显高于肝癌HepG-2细胞(P<0.05);(2)人脐带间充质干细胞的条件培养基和白细胞介素6均能激活STAT3,白细胞介素6中和抗体则明显削弱了人脐带间充质干细胞条件培养基的激活作用;(3)用白细胞介素6中和抗体或AG490抑制STAT3的活性后,肝癌HepG-2细胞的增殖相关基因PCNA、Cyclin D1、Survivin的m RNA表达水平明显下调,其增殖和迁移能力明显下降;(4)结果表明,人脐带间充质干细胞能分泌白细胞介素6激活STAT3信号通路促进肝癌HepG-2细胞的体外增殖和迁移。BACKGROUND：Human umbilical cord mesenchymal stem cel s （hUC-MSCs） can secrete a variety of factors involved in the regulation of tumor proliferation, metastasis and angiogenesis. Probably, interleukin-6 （IL-6） is one of the most important inflammatory factors. OBJECTIVE：To explore the effect of hUC-MSCs on the proliferation and migration of HepG-2 hepatocyte carcinoma cel s via the IL-6/STAT3 signaling pathway. METHODS：IL-6 expression levels in hUC-MSCs and HepG-2 cel s were determined by ELISA. STAT3 and p-STAT3 expression levels were determined by western blot assay. Transcription levels of PCNA, CyclinD1 and STAT3 genes were measured by RT-PCR. HepG-2 cel proliferation was analyzed by flow cytometry and cel counting kit-8 assays. The migration capacity of HepG-2 cel s was evaluated through a scratch test and Transwel assays. RESULTS AND CONCLUSION：The IL-6 level in the hUC-MSCs was significantly higher than that in the HepG-2 cel s （P〈0.05）. Both the hUC-MSC conditioned culture medium and IL-6 could be used for STAT3 activation. The addition of an IL-6 neutralizing antibody significantly weakened the activation of STAT3 in HepG-2 cel s by the hUC-MSCs-conditioned culture medium. In the presence of the IL-6 neutralizing antibody or the STAT3 inhibitor, AG490, the mRNA expression levels of HepG-2 proliferation-related genes （PCNA, CyclinD1 and Survivin） were significantly reduced. The proliferation and migration capacity of HepG-2 cel s were also significantly decreased by this treatment. Taken together, hUC-MSCs can secrete IL-6 to activate the STAT3 signaling pathway, thereby promoting the proliferation and migration of HepG-2 cel s.

关 键 词：干细胞 脐带脐血干细胞 脐带间充质干细胞 HEPG-2细胞 肝癌 白细胞介素6 STAT3 增殖 迁移 云南省自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]