华蟾素对喉癌细胞株的诱导分化研究

The differentiation-inducing effect of cinobutacini on human laryngeal carcinoma cell line Hep-2

目的 研究华蟾素(Cinobutacini,CIN)对喉癌细胞的诱导分化作用。方法 体外培养喉癌细胞(Hep-2)放射性同位素掺人法检测CIN对Hep-2的生长抑制作用,原位杂交方法检测c-myc(细胞核内癌基因)的表达变化。结果 CIN低浓度在24小时内有促进Hep-2细胞分裂、增殖并诱导细胞分化作用;高浓度作用72小时则抑制Hep-2细胞生长。H-脱氧胸苷(H-TdR)掺入量检测提示,CIN短时间作用的Hep-2细胞cpm(每分钟脉冲数)值增高,而长时间作用组则cpm值下降。CIN作用48小时后,Hep-2细胞内c-myc蛋白表达较对照组低,但较平阳霉素组高。结论 中药华蟾素在低浓度和时间相对较短时对人喉癌Hep-2细胞具有诱导分化作用,当作用时间超过72小时,则表现为抑制肿瘤细胞生长作用。

Objective To investigate the differentiation-inducing effect of cinobutacini (CIN)on human laryngeal carcinoma cell line Hep-2. Methods Radioisotope 3H labeled TdR (3H-TdR) was used to determine the growth-inhibiting effect of CIN on Hep-2 by the incorporation method and in situ hybridization technique was taken to detect the changes in the expressive level of oncogene c-myc in these cells, with the morphological changes observed microscopically on the cultured cells in vitro. Results CIN could cause Hep-2 cells dividing to show a tendency of proliferating and a differentiation-inducing effect on the cells cultured together with it for 24 hours at lower concentration,while it showed an inhibitory effect on Hep 2 cultured together for 72 hours at higher concentration. These effects were shown as the content changes of 3H-TdR incorporated to the nucleus of Hep-2 cells. Moreover,the expressive level of c myc protein was significantly lower in the cells cultured together with CIN for 48 hours,than that in the controls, while it was higher than that in the cells cultured with pingyangmyinycin. Conclusions CIN can bring about differentiation-inducing effect on Hep-2 when cultured together with it at lower concentration for a shorter period of time. However, it causes an inhibitory effect on these cells when cultured together for more than 72h.