【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。更多还原

【Abstract】 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 LC3Ⅱ went 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.更多还原