青蒿素通过抑制JAK2/STAT3信号通路诱导肝癌细胞凋亡

Effect of artemisinin inhibiting JAK2/STAT3 pathway on apoptosis of hepatocellular carcinoma cells

目的研究青蒿素对肝癌细胞中JAK2/STAT3信号通路活化的影响,探讨青蒿素抗肿瘤的相关机制。方法选择15只6-8周龄的C57BL/6J小鼠,采用二乙基亚硝胺(diethylnitrosamine,DEN)腹腔注射和CCl4灌胃方法构建小鼠肝癌模型,随机分为PBS组、青蒿素组和青蒿素+p JAK2多肽组,另选5只正常小鼠作为空白组。PBS组和青蒿素组分别腹腔注射PBS及青蒿素(100 mg/kg),隔日作用,3周后处死小鼠;青蒿素+p JAK2多肽组应用p JAK2多肽[100μmol/(L·kg)]隔日作用3次后,再联合青蒿素隔日作用3周,处死小鼠;分离肿瘤组织,Western blot检测肿瘤组织Caspase 3的表达,免疫荧光检测组织中p-JAK2及p-STAT3的表达。结果成功构建小鼠肝癌模型,荷瘤小鼠经青蒿素作用后肿瘤组织Caspase 3蛋白表达显著增加(P<0.001)。同时免疫荧光结果显示青蒿素组肿瘤组织中的p-JAK2及p-STAT3磷酸化水平较之PBS组明显降低;荷瘤小鼠经p JAK2多肽预处理后,青蒿素诱导荷瘤小鼠表达Caspase 3显著降低(P<0.05),差异有统计学意义。结论青蒿素通过抑制JAK2/STAT3信号通路活化诱导肝癌细胞凋亡,进而发挥抗肿瘤作用。

Objective To explore the anti-tumor mechanism of artemisinin by investigating effects of artemisinin on JAK2/STAT signal patway in the hepatocellular carcinoma （HCC）. Methods The C57BL/6J HCC model mice were induced with diethylnitrosamine （DEN） injection and intragastric administration of CC14. Fifteen HCC model mice were randomly divided into PBS group, artemisinin group and polypeptide ＋ artemisinin group, and 5 normal mice were used as control group. The mice were intraperitoneally injected with PBS and artemisinin（109 mg/kg） for three weeks in PBS group and artemisinin group, respectively, and then the mice were killed. Mice in polypeptide ＋ artemisirtin group were killed after intraperitoneal injection of polypeptide 100 μmol/（L·kg） every other day for one week and then treatment with artemisinin for three weeks. After tumor tissues were separated, the expression of caspase 3 in tumor tissues was detected-bylWestern blot, and the expression of p-JAK2 and p-STAT3 was measured with immunofluorescence histo- chemistry. Results The C57BL/6J mice of HCC models were successfully constructed. The expression of Caspase 3 was significantly increased in tumor tissues after treated with artemisinin （P 〈 0. 001 ）, while the p-JAK2 and p-STAT3 were decreased significantly （P 〈 0.05）. Compared with artemisinin group, Caspase 3 expression was decreased significantly after preconditioning with pJAK2 in poly- peptide ＋ artemisinin group （P 〈 0.05 ）. Conclusion Artemisinin may play a vital role in anti-tumor by inducing the apoptosis of hepatocellular carcinoma＂ cells＂ thrugh inhibiting the activation of JAK2/STAT3 signal pathway.