67 kD层粘连蛋白受体对HepG2细胞增殖影响.docVIP

67 kD层粘连蛋白受体对HepG2细胞增殖影响 　【摘要】 目的 探讨67 kD层粘连蛋白受体(67LR)与肝癌细胞生长增殖的关系。 方法 流式细胞术检测67LR 转染的HepG2细胞稳定株细胞膜表面67LR的表达,采用CCK-8试剂盒测定细胞生长曲线,将不同细胞接种于裸鼠皮下,观察其成瘤能力的差异,采用免疫组织化学分析细胞增殖核抗原Ki-67的表达。 结果 67LR高表达的HepG2细胞株其细胞增殖速度明显降低,从第2 d开始有明显差异(Plt;0.01,n=3),在裸鼠体内成瘤能力下降,瘤体平均质量仅为对照组的35.7%,为67LR低表达组的42.9%(Plt;0.01,n=10)。移植瘤的Ki-67表达明显减少。 结论 67LR可以降低肝癌细胞的生长增殖。 【关键词】 受体,层粘连蛋白; 癌,肝细胞; 肿瘤细胞,培养的; 细胞增殖 ABSTRACT: Objective To investigate the effect of 67 kD laminin receptor(67LR) on the proliferation of hepatocellular carcinoma cell. Methods One subclone with high expression of membrance 67LR(LR4), and another subclone with low expression of membrance 67LR(LR6), besides their controll(pcDNA-1) were used. The expression of 67LR was detected by flow cytometric analysis. Cell viability was measured using cell counting Kit-8. Cells were subcutaneously injected into nude mice and the grown tumors were weighted after 6 weeks. The expression of Ki-67 in each tumors was detected by immunohistochemistry. Results The growth of LR4 cells was significantly slower than LR6 and pcDNA-1 since the second day(Plt;0.01,n=3). The average weight of transplanted tumor of LR4 in nude mice were lower than LR6 and pcDNA-1, just as 42.9% of LR6 group and 35.7% of pcDNA-1 group(Plt;0.01,n=10). The expression ki-67 of transplanted tumor of LR4 was also lower than LR6 and pcDNA-1. Conclusion The overexpression of 67LR can suppress the cell proliferation of HepG2 cells. KEY WORDS: receptors,laminin; carcinoma,hepatocellular; tumor cells,cultured; cell proliferation 层粘连蛋白是细胞外基质重要的糖蛋白,与肿瘤细胞表达的相应受体相互作用,影响肿瘤的侵袭和转移。目前已经发现有多种受体可以和层粘连蛋白结合,其中某些受体的表达水平与肝癌发生、发展及预后存在密切关系［1］。Ozaki等应用RT-PCR技术对肝癌中多种层粘连蛋白受体的表达进行研究,发现67 kD层粘连蛋白受体(67 kD laminin receptor,67LR)与整合素α6、β1的表达均显著高于非癌组织［2］。Zheng等发现转移性肝癌67LR无论在蛋白水平还是在mRNA水平均显著高于非转移肝癌［3］,表明67LR在肝癌的转移中起着重要作用。笔者探讨体外转染67LR后细胞膜表面67LR高表达的HepG2细胞生长增殖的变化。 1 材料与方法 1.1 材料 1.1.1 细胞培养 HepG2肝癌细胞(中国科学院上海细胞研究所细胞库),用含10%新生牛血清RPMI-1640培养液(美国Gibco BRL公司),置于体积分数为0.05的CO2、饱和水汽、37 ℃培养箱中培养。转染67LR的HepG2稳定细胞株LR4、LR6;