重组人促血小板生成素对人肝细胞株L02增殖的影响

Effect of recombinant human thrombopoietin on proliferation in human hepatic cell line L02

目的探究重组人促血小板生成素(rhTPO)对健康人肝细胞株L02的影响。方法以健康人肝细胞株L02为研究对象,通过不同剂量rhTPO[对照组(0 U rhTPO)、低剂量组(150 U rhTPO)及高剂量组(750 U rhTPO)]处理L02细胞,观察各组L02细胞的存活率。采用Westernblotting检测各组细胞[A组(0 U rhTPO)、B组(15 U rhTPO)、C组(150 U rhTPO)及D组(1500 U rh TPO)]的细胞外信号调节激酶(ERK)及其磷酸化产物(p-ERK)蛋白的表达水平,并采用荧光定量PCR检测ERK信使RNA(mRNA)表达水平。结果各组L02细胞存活率的比较,差异有统计学意义(F=8.843,P=0.016)。进一步两两比较发现,低剂量组及高剂量组的存活率均显著高于对照组[(107.9±4.0)%、(112.0±1.6)%、(100.0±2.5)%],且高剂量组细胞的存活率更高(P均<0.05)。各组L02细胞间ERK蛋白及ERK mRNA表达水平的比较,差异均无统计学意义(F=1.140,P=0.392;F=0.437,P=0.733),而各组L02细胞间p-ERK蛋白表达水平的比较差异有统计学意义(F=31.400,P<0.001),且B组、C组及D组细胞的p-ERK蛋白表达水平均显著高于A组细胞(P均<0.05),而B组、C组及D组细胞p-ERK蛋白表达水平两两比较,差异均无统计学意义(P均>0.05)。结论rhTPO可通过激活ERK促进L02细胞数量增殖。

Objective To investigate the effect of recombinant human thrombopoietin(rhTPO) on healthy human hepatic cell line L02. Methods The survival rate of L02 cells was observed among a control group(0 U rhTPO), a low-dose group(150 U rhTPO) and a high-dose group(750 U rhTPO) according to different doses of rhTPO. The expression levels of extracellular signal regulated kinase(ERK) and phosphorylation-ERK(p-ERK) protein were detected by Westernblotting, and the expression level of ERK messenger RNA(mRNA) was examined by fluorogenic quantitative PCR in the L02 cells among group A(treated with 0 U rhTPO), group B(treated with 15 U rhTPO), group C(treated with 150 U rhTPO) and group D(treated with 1 500 U rhTPO).Results The survival rate of L02 cells was significantly different among the three groups(F =8.843, P = 0.016). It was much higher in the low-dose group and high-dose group than in the control group, and highest in the high-dose group [(107.9 ± 4.0)%,(112.0 ± 1.6)%,(100.0 ± 2.5)%;all P < 0.05]. However, there were no statistically significant differences in the expression levels of ERK protein and ERK mRNA among the four groups(F = 1.140, P = 0.392;F = 0.437, P =0.733). Besides, the expression level of p-ERK protein was significantly different among the four groups(F = 31.400, P < 0.001). It increased obviously in group B, group C and group D as compared with group A(all P < 0.05), while no significant differences were noted among group B, group C and group D(all P > 0.05). Conclusion rhTPO can promote the proliferation of L02 hepatocytes by activating ERK.