p-PKCδ通过IL-6/STAT3通路促进GMA诱导的恶性转化16HBE细胞增殖并抑制细胞凋亡  

Phosphorylated PKCδ promotes GMA-induced proliferation and inhibits apoptosis in malignantly transformed 16HBE cells through the IL-6/STAT3 pathway

作　　者：金惠萍 王全凯[1,2] 李昕苇 崔旭芳 顾轶婷 乌翰宝栎尔 康同影[1] 张林媛[1,2] 许建宁[1,2] JIN Hui-ping;WANG Quan-kai;LI Xin-wei;CUI Xu-fang;GU Yi-ting;WUHAN Bao-lier;KANG Tong-ying;ZHANG Lin-yuan;XU Jian-ning(National Institute of Occupational Health and Poison Control,Chinese Center for Disease Control and Prevention,Bejing 100050,China;State Key Laboratory of Trauma and Chemical Poisoning,Chinese Center for Disease Control and Prevention,Beijing 100050,China)

机构地区：[1]中国疾病预防控制中心职业卫生与中毒控制所,北京100050 [2]中国疾病预防控制中心创伤与化学中毒全国重点实验室,北京100050

出　　处：《毒理学杂志》2024年第6期407-413,425,共8页Journal of Toxicology

基　　金：国家自然科学基金(81673221)。

摘　　要：目的探讨甲基丙烯酸缩水甘油酯(glycidyl methacrylate,GMA)诱导16HBE细胞发生恶性转化的关键分子机制,是否与磷酸化的PKCδ(p-PKCδ)表达增加相关。方法基于8μg/ml GMA诱导的恶性转化16HBE细胞模型;使用Western blot确认p-PKCδ的表达情况;使用p-PKCδ特异性抑制剂GO6983抑制p-PKCδ的表达,通过Western blot、qPCR技术分析p-PKCδ、IL-6/磷酸化转录3激活剂(STAT3)通路及凋亡相关蛋白Caspase-3的蛋白和转录表达水平;同时,使用细胞划痕实验检测细胞的迁移能力。结果Western blot和qPCR结果表明,与DMSO对照组相比,p-PKCδ的表达增加(t=31.14,P<0.05);p-PKCδ抑制后p-STAT3的蛋白表达呈现降低趋势(t=11.05,P<0.05),同时凋亡蛋白Caspase-3表达明显增加(t=12.95,P<0.05),同时转录水平也呈现相同变化趋势;与对照组相比,p-PKCδ抑制组划痕实验提示细胞迁移能力减弱。结论p-PKCδ与GMA诱导的16HBE细胞恶性转化过程密切相关,p-PKCδ可通过激活IL-6/STAT3通路,促进GMA诱导的恶性转化16HBE细胞的增殖并抑制其凋亡。本研究的结果将有助于阐明GMA的致癌机制。Objective To investigate whether the key molecular mechanism of GMA-induced malignant transformation of 16HBE cells is associated with increased expression of phosphorylated PKCδ(p-PKCδ).Methods A malignant transformation 16HBE cell model exposed to 8μg/ml GMA;its expression was confirmed using Western blot;p-PKCδ expression was inhibited using the p-PKCδ-specific inhibitor GO6983,and p-PKCδ,IL-6/phosphorylated transcription 3 activator(STAT3)pathway and protein and mRNA expression of apoptosis-related protein Caspase-3 were analyzed by Western blot and qPCR methods.Meanwhile,cell migration ability was detected using cell wound healing assay.Results Western blot result showed that the expression of p-PKCδ was increased compared with the DMSO control group(t=31.14,P<0.05);the expression of apoptosis-related protein Caspase-3 was increased in the p-PKCδ-inhibited group(t=12.95,P<0.05),whereas p-STAT3 showed decreased expression(t=11.05,P<0.05),as well as the transcript levels showed the same change trend;compared with the control group,the wound healing assay of p-PKCδinhibition group indicated that the cell migration ability was reduced.Conclusion p-PKCδ is closely related to the malignant transformation process of GMA-induced 16HBE cells.p-PKCδ can promote the proliferation and inhibit the apoptosis of GMA-induced malignant transformed 16HBE cells by activating the IL-6/STAT3 pathway.The result of this study will help to elucidate the oncogenic mechanism of GMA.

关 键 词：甲基丙烯酸缩水甘油酯 细胞恶性转化 p-PKCδ IL-6/STAT3 

分 类 号：R994.6[医药卫生—毒理学] R114[医药卫生—药学] R730.2