山东医药2026，Vol.66Issue(4)：21-25,5.DOI:10.3969/j.issn.1002-266X.2026.04.005
基于miR-34b-3p/PINK1/Bcl-2信号通路探讨蓝萼甲素调控肝癌高转移细胞系HCCLM3自噬与凋亡的机制
Mechanism of glaucocalyxin A regulating autophagy and apoptosis in highly metastatic hepatocellular carcinoma cell line HCCLM3 based on miR-34b-3p/PINK1/Bcl-2 signaling pathway
摘要
Abstract
Objective To investigate the mechanism by which glaucocalyxin A(GLA)regulates mitophagy and apop-tosis in hepatocellular carcinoma cells with a focus on the miR-34b-3p/PINK1/Bcl-2 signaling pathway.Methods The highly metastatic hepatocellular carcinoma cell line HCCLM3 was used.Cells were divided into the control group and GLA-treated groups(2.5,5,and 10 μg/mL).Cells in the control group were cultured under routine conditions,while cells in the GLA groups were treated with 2.5,5,and 10 μg/mL GLA,respectively.The expression of miR-34b-3p was detected by quantitative real-time PCR(qPCR).The protein expression levels of PINK1,Parkin,LC3,Bax,Bcl-2,and Cleaved Caspase-3 were determined by Western blotting.The apoptosis was analyzed by flow cytometry.The intracellular localization of PINK1 was observed by fluorescence microscopy.The interaction between PINK1 and Parkin was assessed by co-immu-noprecipitation.Cellular autophagy and apoptosis were observed by transmission electron microscopy.Results Compared with the control group,no statistically significant differences were found in the expression of miR-34b-3p,the relative expres-sion levels of mitophagy-and apoptosis-related proteins(PINK1,Parkin,LC3Ⅱ/LC3Ⅰ,Bax,Bcl-2,Cleaved Caspase-3),or the apoptosis rate in the GLA 2.5 μg/mL group(all P>0.05);in the GLA 5 μg/mL group,miR-34b-3p expression showed no statistically significant difference(P>0.05),whereas the expression levels of PINK1 and Parkin increased,the LC3Ⅱ/LC3Ⅰratio was elevated,Bax and Cleaved Caspase-3 expression increased,Bcl-2 expression decreased,and the apoptosis rate in-creased(all P<0.05),and transmission electron microscopy revealed mitochondrial swelling and an increased number of au-tophagosomes;in the GLA 10 μg/mL group,miR-34b-3p expression significantly increased,PINK1 expression decreased a-long with reduced intracellular fluorescence intensity,Parkin and Bcl-2 expression decreased,while Bax and Cleaved Caspase-3 expression increased,and the apoptosis rate increased(all P<0.05),and typical apoptotic morphology was ob-served under transmission electron microscopy.Compared with the GLA 5 μg/mL group,the binding level of PINK1 and Par-kin was reduced in the GLA 10 μg/mL group.Conclusion GLA may regulate autophagy and apoptosis in HCCLM3 cells by modulating the miR-34b-3p/PINK1/Bcl-2 signaling pathway,and its effects are dose-dependent.关键词
蓝萼甲素/肝癌/miR-34b-3p/PINK1/Bcl-2信号通路/自噬/凋亡Key words
glaucocalyxin A/hepatic carcinoma/miR-34b-3p/PINK1/Bcl-2 signaling pathway/autophagy/apoptosis分类
医药卫生引用本文复制引用
朱琳琳,张秀..基于miR-34b-3p/PINK1/Bcl-2信号通路探讨蓝萼甲素调控肝癌高转移细胞系HCCLM3自噬与凋亡的机制[J].山东医药,2026,66(4):21-25,5.基金项目
河南省科技攻关项目(252102310079) （252102310079）
河南省科技研发计划联合基金项目(252103810345). （252103810345）
