[关键词]
[摘要]
目的观察苦参碱对肝癌SMMC-7721细胞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,MTT法检测苦参碱对肝癌SMMC-7721细胞株增殖的影响,RT-PCR法检测苦参碱对SMMC-7721细胞stat3、stat5 mRNA表达的影响,Western blotting法检测苦参碱对肝癌SMMC-7721细胞STAT3、STAT5、P-STAT3、P-STAT5蛋白表达的影响。结果苦参碱对肝癌细胞增殖的抑制率呈剂量和时间依赖性。苦参碱能下调stat3、stat5 mRNA表达水平(P<0.05、0.01),降低STAT3、STAT5、P-STAT3、P-STAT5蛋白表达量(P<0.05、0.01)。AG490作用于SMMC-7721细胞后,stat3、stat5 mRNA和STAT3、STAT5蛋白的表达量与对照组比较无显著差异(P>0.05),P-STAT3、P-STAT5蛋白表达量与对照组比较显著降低(P>0.05、0.01)。与AG490组比较,AG490+苦参碱组stat3、stat5 mRNA的表达量显著降低(P<0.05),STAT3、STAT5蛋白表达量显著降低(P>0.05),P-STAT3、P-STAT5蛋白表达量无显著差异(P>0.05)。与苦参碱组比较,AG490+苦参碱组stat3、stat5 mRNA的表达量无显著差异(P>0.05),STAT3、STAT5、P-STAT3、P-STAT5蛋白表达量无显著差异(P>0.05)。结论苦参碱能下调stat3、stat5 mRNA表达水平,因而能降低STAT3、STAT5蛋白表达水平,抑制细胞JAK-STAT信号转导通路,从而抑制肝癌SMMC-7721细胞增殖。
[Key word]
[Abstract]
Objective To investigate the effect of matrine on JAK-STAT signaling transduction pathway in SMMC一7721 cell line.Methods Treated with matrine and AG490(the inhibitor of JAK-STAT signaling transduction)。the inhibitory effect of matrine on SMMC一7721 cell proliferation wasdetected by MTT.The mRNA expression of stat3 and stat5 in SMMC一7721 cell line was assessed withRT-PCR.The protein expression of STAT3,STAT5,P-STAT3 and P-STAT5 in SMMC一7721 cell linewas detected by Western blotting.Results Matrine could significantly inhibit the proliferation of SMMC一7721 cells.The proliferation rate was in fl dose-and time-dependent manner.The mRNA expression ofstat3 and stat5 in SMMC一7721 cell with matrine was significantly Iower than that in the control group(PO.05),and so was the protein expression of STAT3 and STAT5 (P>0.05).But P-STAT3 and P-STAT5 protein expression was significantly lower than that in thecontrol group(P<0.05 or P0.05).There was no signi-fieant difference of the mRNA expression of stat 3 and stat 5 between matrine plus AG490 group andmatrine group(P>O.05),and so was STAT3,STAT5,P-ERK,P-STAT3 and P-STAT5 protein expre-ssion(P>O.05).Conclusion Matrine could significantly down-regulate the mRNA expression of stat3and stat5 and protein expression of STAT3 and STAT5 in SMMC一7721 cells.So matrine could inhibit thesignaling transduction pathway of JAK-STAT and inhibit the proliferation of SMMC一7721 cells.
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[基金项目]
河北省科技领军人才资金资助项目(06547008D-4)