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目的探讨肿瘤相关巨噬细胞(TAMs)通过JAK2/STAT3途径对肝癌细胞凋亡的影响。方法培养人单核巨噬细胞(THP1),加入佛波酯(PMA)及白介素4(IL-4)诱导分化为TAMs,RT-PCR检测TAMs表面分子标记物CD206和CD163的表达;TAMs与肝癌细胞(Hep G2、SMMC7721)在Transwell小室内共培养48h,通过western blot方法检测凋亡相关蛋白Caspase 3、抗凋亡蛋白Bcl-2、信号通路JAK2/STAT3以及p-STAT3的表达;加入JAK2/STAT3通路特异性阻断剂AG490进行阻断后,检测相关凋亡蛋白、抗凋亡蛋白表达情况以及JAK/STAT信号通路活化情况。结果人单核巨噬细胞(THP1)经PMA及IL-4分化诱导72h,RT-PCR检测其细胞表面CD206和CD163的表达显著增加(P<0.05);肝癌细胞(Hep G2、SMMC7721)与TAMs共培养48h,Western Blot检测肿瘤细胞凋亡相关蛋白Caspase3、Bax表达明显降低,抗凋亡蛋白Bcl-2以及信号通路JAK2、STAT3、p-STAT3表达显著升高;AG490阻断JAK2/STAT3通路后,加AG490组肿瘤细胞p-STAT3表达降低,凋亡相关蛋白Caspase3、Bax的表达明显升高,抗凋亡相关蛋白Bcl-2的表达显著降低;结论肿瘤相关巨噬细胞(TAMs)通过激活JAK2/STAT3信号途径调控肝癌细胞的凋亡。
Objective To investigate the effect of tumor-associated macrophages (TAMs) on apoptosis of hepatocellular carcinoma cells by JAK2 / STAT3 pathway. METHODS: Human monocyte-macrophage (THP1) cells were cultured and transfected with phorbol ester (PMA) and interleukin-4 (IL-4) to differentiate into TAMs. The expression of CD206 and CD163 on TAMs were detected by RT- (Hep G2 and SMMC7721) were co-cultured in Transwell chamber for 48h. The expressions of Caspase 3, anti-apoptotic protein Bcl-2, JAK2 / STAT3 and p-STAT3 were detected by western blot. After blocking with AG490, a specific inhibitor of STAT3 pathway, the expression of anti-apoptotic protein and anti-apoptotic protein and the activation of JAK / STAT signaling pathway were detected. Results The expression of CD206 and CD163 on the surface of THP1 cells induced by PMA and IL-4 was significantly increased (P <0.05) by RT-PCR for 72 hours. Compared with TAMs (Hep G2 and SMMC7721) After co-cultured for 48h, the expressions of Bcl-2 and JAK2, STAT3 and p-STAT3 were significantly increased by Western Blot, while the expressions of Caspase3 and Bax were significantly decreased after AG490 blocking JAK2 / STAT3 pathway , While the expression of p-STAT3 in AG490 group decreased, the expressions of apoptosis-related proteins Caspase3 and Bax were significantly increased, and the expression of anti-apoptosis related protein Bcl-2 was significantly decreased. Conclusions Tumor-associated macrophages (TAMs) / STAT3 signaling pathway regulates hepatoma cell apoptosis.