大肠癌细胞增殖中HGF/SF作用的观察

The function of HGF/SF in the proliferation of colorectal cancer cells

目的研究肝细胞生长因子/离散因子(HGF/SF)在诱导大肠癌细胞增殖中的作用。方法采用W estern B lot方法,检测HGF的受体c-met在受检大肠癌细胞株Caco-2,Colo3 2 0中的表达;观察Caco-2,Colo3 2 0中HGF/SF活化p 4 2/p 4 4MAPK和p 3 8MAPK的动态变化;应用[3H]-TdR,MTT方法观察p 4 2/p 4 4MAPK和p 3 8MAPK传导通路阻滞剂PD 9 8 0 5 9和SB 2 0 3 5 8 0对HGF/SF诱导的大肠癌细胞增殖的抑制作用。结果(1)c-met在Caco-2和Colo3 2 0中有表达。(2)HGF/SF激活p 4 2/p 4 4MAPK,p 3 8MAPK:2 0 ng/mL的HGF/SF处理细胞,p 4 2/p 4 4MAPK磷酸化在1 0m in达高峰(2.2 8±0.0 1);p 3 8MAPK变化与之相似(2.2 5±0.0 1)。(3)HGF/SF诱导大肠癌细胞的DNA合成增加依赖于p 4 2/p 4 4MAPK的激活,在2 4 h时点分别以2 0 ng/m lHGF/SF,不同浓度(1μmol/L,5μmol/L,1 0μmol/L)的PD 9 8 0 5 9和SB 2 0 3 5 8 0处理细胞,HGF/SF使胸腺啶吸收增加(P<0.0 1);PD 9 8 0 5 9以浓度依赖性抑制胸腺啶的吸收(P<0.0 1)。(4)HGF促进Caco-2细胞的增殖,而PD 9 8 0 5 9对这种增殖有抑制作用。结论HGF激活大肠癌细胞Caco-2和Colo3 2 0中p 4 2/p 4 4MAPK和p 3 8MAPK;p 4 2/p 4 4MAPK参与HGF/SF诱导的大肠癌细胞Caco-2有丝分裂;HGF促进大肠癌细胞Caco-2增殖;HGF/SF和p 4 2/p 4 4MAPK在大肠癌细胞中发挥作用可能有细胞选择性。

Objective To study the function of hepatocyte growth factor/scatter factor （HGF/SF） in the proliferation of colorectal cancer cells. Methods The expression of c-met, the receptor of HGF, was detected in Caco-2 and Colo320 cell lines by Western blot. The activation of p42/p44MAPK and p38 MAPK induced by HGF in these two cell lines was observed. Observation of the effect of the inhibitor of p42 / p44 MAPK （ PD 98059 ） , p 38 MAPK （ SB 203580 ） on the inhibition of HGF - induced proliferation of Caco-2 and Colo320 cells were made by Using [^3H ] -TdR, MTY assay. Results （ 1 ） Both cell lines expressed the c-met. （2）HGF activated p42/p44 MAPK and p38 MAPK, and 20ng/ml HGF treated cells showed maximum activity in both to be within 10min. （ p42/p44MAPK,2.28 ± 0.01 ;p38MAPK, 2.25± 0.01 ） . （ 3 ） HGF was found to significantly increase [ ^3 H ] thymidine incorporation （ P 〈 0. 01 ） , when cells were pretreated with inhibitors of p42/p44MAPK （PD98059） in different doses （1 μmol/L ,5 μLmol/ L, 10μmol/L） , HGF-induced thymidine uptake was suppressed in a dose-dependent manner （ P 〈 0.01 ）. （4） MTY assay found HGF enhanced the proliferation of Caco-2 , however, PD98059 inhibited this proliferation. Conclusions The results demonstrate that HGF activates MAPK in both colorectal carcinoma cell lines, and p42/p44MAPK take part in the mitosis of the Caco-2 cells, while HGF promotes the proliferation of Caco-2 cells. The function of HGF/SF and p42/p44MAPK in colorectal cancer cells may have cellular selection.