磷酸化胰岛素样生长因子结合蛋白-1对肝癌细胞体外生长抑制作用

Inhibited effect of phosphorylated insulin-like growth factor binding protein-l on hepetoma Cells in vitro

目的探讨胰岛素样生长因子结合蛋白－１（ＩＧＦＢＰ－１）对肝癌细胞选择性生长抑制作用和对胰岛素样生长因子Ⅱ（ＩＧＦⅡ）调解机理。方法采用从孕妇羊水中纯化的磷酸化结合蛋白－１（ＰＢＰ－１）和非磷酸化结合蛋白－１（ｎｐＢＰ－１），自肝细胞癌实体瘤和肝性腹水标本中获取瘤细胞。应用ＭＴＴ试验，地高辛标记的原位杂交和膜上斑点杂交试验，分别定性和定量测定各实验组中肝癌细胞生长和ＩＧＦⅡｍＲＮＡ表达的动态变化。结果实验表明，ＰＢＰ－１组细胞与对照组相比，同一时间细胞生长趋缓，于培养第２天即呈下降趋势（Ｐ＜０．０５）；原位杂交显示各组细胞在整个培养期间，均有ＩＧＦⅡｍＲＮＡ阳性信号。膜上斑点杂交经光密度扫描表明，ｐＢＰ－１组细胞的ＩＧＦⅡｍＲＮＡ表达平均光密度值与对照组和其他各组比较差异均有显著性（Ｐ＜０．０１），ＰＰＢＰ－１及其他组则无明显差异。结论ｐＢＰ－１在体外可抑制肝癌细胞的生长；ＩＧＦⅡｍＲＮＡ表达可能与ｍＢＰ—１的调节抑制作用有关。

Objective To elucidate the inhibited effect and regUlated mechanism of IGFBp-l on thecarcinoma cells. Methods The recombinant human IGFn (rhIGF ), phosphorylated and nonphosphorylated binding protein-l(pBp-l and npBp-l) that were purified from pregnant woman's amnoitic fluidwere used to cultivate cancer cells from 15 cases with hepatocellular carcinoma and ascites. The cellcounting and MTT methods were uest for detennining the dynamics change of proliferation on HCC, andthe in situ or dot hybridhation with Dboxin label used for the expression of IGF mRNA qualitativelyand quantitatively at different times. Results The cells grew more slowly, in pBp-l group, than incontrol group at the same time, which begun to decrease at tWo dsys(P<0.05) by MTT. In situ hybridization showed that there are hybridization Signal of IGFn mRNA in all groups during the cultrue, andscanning dot hybridization showed the OPTMD of IGF mRNA in pBp-l group was statistically statisticallyfrom control and other groups (P<0.01), while in npBp-l group there was no significant effect comperedwith other groups (PM0.05). Conclusion The pBp-l can inhibit the growth of hepatoma cells and theexpression of IGF mRNA, which may be regulated and inhibited by pBp-l.