苦参碱对SMMC-7721细胞MAPK、JAK-STAT信号通路的影响

doi:10.3971/j.issn.1000-8578.3413

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

摘要

摘要: 目的观察苦参碱对肝癌细胞SMMC-7721 MAPK通路和JAK—STAT通路的影响。方法苦参碱和（或）JAN-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、STAT5 mRNA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、STAT5 mRNA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Pd0．01）。与AG490组比较，AG490＋苦参碱组ERK、STAT3、STAT5 mRNA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、STAT5 mRNA的表达量差异无统计学意义（P〉0．05），ERK、STAT3、STAT5、P-ERK、P-STAT3、P-STAT5蛋白袁达量差异无统计学意义（P〉0．05）。结论苦参碱能下调ERK、STAT3、STAT5 mRNA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。

Abstract: Objective 　To investigate the effect of mat rine on MAPK and JA K2STAT signaling t ransduc2 tion pathway in SMMC27721 cell line. Methods 　Treated with mat rine and A G490 (the inhibitor of JA K2 STAT signaling t ransduction), the mRNA expression of ERK, STAT3 and STAT5 in SMMC27721 cell line were assessed with RT2PCR. The protein expression of ERK, STAT3, STAT5, P2ERK, P2STAT3 and P2STAT5 in SMMC27721 cell line were detected by Western blotting. Results 　The mRNA expres2 sion of ERK, STAT3 and STAT5 in SMMC27721 cell with mat rine was significantly lower than those in cont rol group ( P < 0. 05 or P < 0. 01) and so did the protein expression of ERK, STAT3, STAT5, P2 ERK, P2STAT3 and P2 STAT5 ( P < 0. 05 or P < 0. 01) . There were no significant difference of the mR2 NA expression of ERK, STAT3 and STAT5 between A G490 and cont rol group ( P > 0. 05), and so did the protein expression of ERK, STAT3 and STAT5 ( P > 0. 05) . But P2ERK, P2STAT3 and P2 STAT5 protein were significantly lower than those in cont rol group ( P < 0. 05 or P < 0. 01) . After t reated with mat rine plus AG490, the mRNA 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 P2ERK, P2STAT3 and P2 STAT5 protein between mat rine plus A G490 group and A G490 group ( P > 0. 05) . There were no significant difference of the mRNA expression of ERK, STAT3 and STAT5 between mat rine plus A G490 group and mat rine group ( P > 0. 05), and so did ERK, STAT3, STAT5, P2ERK, P2STAT3 and P2 STAT5 protein ( P > 0. 05) . Conclusion 　Mat rine could significantly down2regulate the mRNA expression of ERK, STAT3, STAT5 and protein expression of ERK, STAT3 and STAT5 in SMMC27721 cells. So mat rine could inhibit the signaling t ransduction pathway of MAPK and JA K2STAT and inhibit the proliferation of SMMC27721 cells.

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