蛋白激酶C和N a^+/H^+交换在EGF促肝癌细胞生长中的作用

Effect of protein kinase C and Na^+/H^+ exchange on the proliferation of hepatoma cells induced by epidermal growth factor

目的研究蛋白激酶 C 和 Na^+/H^+交换在表皮生长因子(EGF)促人肝癌细胞生长中的作用.方法采用无血清 RPM11640培养基培养人肝癌细胞SMMC7721,给以 EGF 10^(-9)mol·L^(-1),以~3H-7hymidine(~3H-TdR)掺入法,检测肝癌细胞 DNA 合成的速率.并分别用蛋白激酶 C 阻滞剂 H-7 50μml·L^(-1)和 Na^+/H^+交换阻滞剂amiloride 0.1 mmol·L^(-1)阻断 EGF 对肝癌细胞的促生长作用.结果 EGF 10^(-9)mol·L^(-1)可显著促进肝癌细胞的生长,掺入值为(1880±281)·min^(-1)·孔^(-1),与对照组掺入值(1353±175)·min^(-1).孔^(-1)比较,差异有显著性(P<0.05).单纯 H-7或amiloride,掺入值分别为(1179±150)·min^(-1)·孔^(-1)和(1392±152)·min^(-1)·孔^(-1),与对照组差异无显著性(P>0.05),对肝癌细胞的生长无明显影响;而 H-7,amiloride 与 EGF 合用,掺入值分别为(1241±147)·min^(-1)·孔^(-1)和(1380±189)·min^(-1)·孔^(-1),与 EGF 组相比差异有显著性(P<0.01,P<0.05).结论 EGF 能显著促进肝癌细胞的生长,蛋白激酶 C 和 Na^+/H^+交换在 EGF 的促肝癌细胞生长中起关键作用.

AIM To study the effect of protein kinase C and Na^+/H^+ exchange on the proliferation of human hepatoma cell line SMMC7721 induced by epidermal growth factor(EGF). METHODS Human hepatoma cells SMMC7721 were cultured in RPMI1640 serum-free medium.EGF 10^(?)mol· L^(-1)was added,and the DNA synthesis rate was checked by the method of^3H-Thymidine(~3H-TdR)incorporation.Then protein kinase C inhibitor H-7 and Na^+/H^+ exchanger inhibitor amiloride were used respectively to block the effect of EGF on the hepatoma cells. RESULTS The proliferation of hepatoma cell line was stimulated by EGF 10^(-9)mol·L^(-1)significantly,the incorporations were(1880±281)·min^(-1).well^(-1)and(1353± 175)·min^(-1)·well^(-1)respectively(P<0.05).Protein kinase C inhibitor H-7 50μmol·L^(-1)or Na^+/H^+ exchanger inhibitor amiloride 0.1mmol·L^(-1) was used lonely,the incorporations were(1179±150)·min^(-1)·well^(-1)and(1392±152)·min^(-1). well^(-1)respectively,with no significant difference compared with the control(P>0.05),and no effect on the growth of hepatoma cells.But when H-7 or amiloride used with EGF, the incorporation was(1241±147)·min^(-1)·well^(-1)and(1380 ±189)·min^(-1)·well^(-1),had significant differences compared with EGF group(P<0.01,P<0.05). CONCLUSION The proliferation of hepatoma cell line was stimulated by EGF 10^(-9)mol·L^(-1).Protein kinase C and Na^+/ H^+ exchange play a critical role in the proliferation of hepatoma cells induced by EGF.