苦参碱对SMMC-7721细胞MAPK、JAK-STAT信号通路的影响

Effect of Matrine on Signaling Transduction of MAPK and JAK-STAT in SMMC-7721 Cell Line

目的观察苦参碱对肝癌细胞SMMC-7721MAPK通路和JAK-STAT通路的影响。方法苦参碱和(或)JAK-STAT途径特异性抑制剂AG490培养肝癌细胞SMMC-7721,RT-PCR法检测苦参碱对SMMC-7721细胞ERK,STAT3、STAT5mRNA的影响,Western blott法检测苦参碱对肝癌SMMC-7721细胞ERK、STAT3、STAT5、P-ERK、P-STAT3、P-STAT5蛋白表达的影响。结果苦参碱能下调ERK,STAT3、STAT5mRNA表达水平,降低ERK、STAT3、STAT5、P-ERK、P-STAT3、P-STAT5蛋白表达量(P<0.05或P<0.01)。AG490作用于SMMC-7721细胞后,ERK,STAT3、STAT5mRNA和ERK、STAT3、STAT5蛋白的表达量与对照组比较差异无统计学意义(P>0.05),P-ERK、P-STAT3、P-STAT5蛋白表达量与对照组比较显著降低(P<0.05或P<0.01)。与AG490组比较,AG490+苦参碱组ERK、STAT3、STAT5mRNA的表达量显著降低(P<0.05),ERK、STAT3、STAT5蛋白表达量显著降低(P<0.05),P-ERK、P-STAT3、P-STAT5蛋白表达量差异无统计学意义(P>0.05)。与苦参碱组比较,AG490+苦参碱组ERK,STAT3、STAT5mRNA的表达量差异无统计学意义(P>0.05),ERK、STAT3、STAT5、P-ERK、P-STAT3、P-STAT5蛋白表达量差异无统计学意义(P>0.05)。结论苦参碱能下调ERK、STAT3、STAT5mRNA表达水平,因而能降低ERK、STAT3、STAT5蛋白表达水平,抑制细胞信号转导通路,从而抑制肝癌细胞增殖。

Objective To investigate the effect of matrine on MAPK and JAK-STAT signaling transduction pathway in SMMC-7721 cell line. Methods Treated with matrine and AG490（the inhibitor of JAK- STAT signaling transduction） , the rnRNA expression of ERK, STAT3 and STAT5 in SMMC-7721 cell line were assessed with RT-PCR. The protein expression of ERK, STAT3, STAT5, P-ERK, P-STAT3 and P-STAT5 in SMMC-7721 cell line were detected by Western blotting. Results The rnRNA expression of ERK, STAT3 and STAT5 in SMMC-7721 cell with matrine was significantly lower than those in control group （P〈0. 05 or P〈0. 01 ） and so did the protein expression of ERK, STAT3, STAT5, P- ERK, P-STAT3 and P- STAT5 （P〈0. 05 or P〈0.01 ）. There were no significant difference of the mRNA expression of ERK, STAT3 and STAT5 between AG490 and control group （P〉0. 05） , and so did the protein expression of ERK, STAT3 and STAT5 （P〉0.05）. But P-ERK, P-STAT3 and P- STAT5 protein were significantly lower than those in control group （P〈0. 05 or P〈0. 01 ）. After treated with rnatrine plus AG490, the rnRNA expression of ERK, STAT3 and STAT5 were significantly lower than those in AG490 group （P〈0. 05） and so did the protein expression of ERK, STAT3, STAT5 （P〈0. 05）. There were no significant difference of the P-ERK, P-STAT3 and P- STAT5 protein between matrine plus AG490 group and AG490 group （P〉0.05）. There were no significant difference of the rnRNA expression of ERK, STAT3 and STATS between matrine plus AG490 group and matrine group （P〉0. 05）, and so did ERK, STAT3, STAT5, P-ERK, P-STAT3 and P-STAT5 protein （P〉0. 05）. Conclusion Matrine could significantly down-regulate the rnRNA expression of ERK, STAT3, STAT5 and protein expression of ERK, STAT3 and STAT5 in SMMC-7721 cells. So matrine could inhibit the signaling transduction pathway of MAPK and JAK-STAT and inhibit the proliferation of SMMC-7721 cells.