摘要
目的研究B7同源蛋白4/含V-SET域T细胞激活抑制因子1(B7-H4/VTCN1)对肝癌细胞凋亡和自噬的影响及其可能的信号通路。方法采用小干扰RNA技术敲低Huh7细胞B7-H4,CCK-8法检测细胞增殖,流式细胞术检测细胞凋亡;Western blot法检测细胞裂解型胱天蛋白酶3(c-caspase-3)、B细胞白血病/淋巴细胞瘤分子2(Bcl2)、微管相关蛋白1轻链3(LC3)、P62和c-Jun氨基末端激酶(JNK)及磷酸化的JNK(p-JNK)蛋白水平;单丹磺酰尸胺(MDC)法检测自噬体。结果敲低Huh7细胞B7-H4后,肝癌细胞凋亡和自噬增加,细胞增殖降低;c-caspase-3、LC3Ⅱ蛋白增加,Bcl2、P62蛋白降低;JNK的磷酸化被抑制,可见自噬体形成。结论敲低B7-H4促进Huh7细胞凋亡和自噬,可能与抑制JNK的磷酸化有关。
Objective To investigate the effects of co-stimulatory molecule B7-H4/VTCN1 on apoptosis and autophagy of hepatocellular carcinoma(HCC)cells and the potential signaling pathways.Methods After Huh7 cells were treated by B7-H4 siRNA,CCK-8 assay was used to detect the cell proliferation.Cell apoptosis was measured by flow cytometry.The protein expression levels of cleaved caspase-3(c-caspase-3),Bcl2,LC3,P62,JNK and phosphorylated JNK(p-JNK)were examined by Western blot analysis.The autophagosome was observed by monodansylcadaverine(MDC)assay.Results After the knockdown of B7-H4,the apoptosis and autophagy of HCC cells increased,and cell proliferation decreased.Moreover,the expression levels of c-caspase-3 and LC3Ⅱwent up,while the expression levels of Bcl2 and P62 went down.Furthermore,the phosphorylation of JNK was also inhibited,and autophagosome was visible.ConclusionKnockdown of B7-H4 promotes the apoptosis and autophagy in HCC cells,which may be related to the inhibited phosphorylation of JNK.
作者
郝团团
雷登亮
胡刚理
廖锐
罗放
HAO Tuantuan;LEI Dengliang;HU Gangli;LIAO Rui;LUO Fang(Department of Hepatobiliary Surgery,First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2020年第7期603-608,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金(81372481)。
