目的探讨弯齿琵甲含药血清对宫颈癌HeLa细胞增殖的影响及其抗肿瘤的作用机制。方法采用血清药理学方法制备不同浓度的弯齿琵甲含药血清,MTT法检测不同浓度含药血清对宫颈癌HeLa细胞增殖的影响;显微镜下观察W right's-G iem sa染色和吖啶橙-EB染色后细胞的形态学变化;DNA琼脂糖凝胶电泳检测凋亡细胞DNA断裂情况。结果宫颈癌HeLa细胞经不同剂量的弯齿琵甲含药血清分别作用24、48 h后,细胞增殖明显受到抑制,且呈时效、量效关系;倒置显微镜下观察可见细胞膜皱缩,细胞核固缩,核染色质边缘化,并出现凋亡小体;荧光显微镜下可见早期凋亡细胞呈黄绿色球状,晚期凋亡细胞呈红色球状;凋亡细胞经琼脂糖凝胶电泳可观察到DNA梯形条带。结论弯齿琵甲含药血清对宫颈癌HeLa细胞增殖具有明显的抑制作用,其作用机制与诱导细胞凋亡相关。
Objective To investigate the inhibitory effect of serum containing blaps femoralis Fischer-Waldheim on proliferation of HeLa cell line in vitro and its probable anti-tumor mechanism.Methods Rabbit serum containing drug was prepared by using serologic phamacology method,in which,Blaps femoralis Fischer-Waldheim was contained.MTT assay was used to evaluate the inhibition effect on HeLa cell growth.The apoptosis morphological changes were observed under the microscope with Wright's-Giemsa staining and acridine orange-EB staining of HeLa cells.The DNA agarose electrophoresis was performed to determine apoptosis and biochemical changes of HeLa cells.Results The different concentrations of serum containing Blaps femoralis Fischer-Waldheim had significant inhibitory effects on HeLa cells at 24 hours and 48 hours,which showed in a time-and dose-dependent manner.With inverted microscope,the HeLa cell showed the typical apoptosis morphological changes,such as cell membrane shrinkage,condensation of chromosomes and apoptotic bodies with complete membranel with fluorescence microscope,viable apoptotic cells were showed round in relly,non-viable apoptotic cells were showed round in red;Furthermore,the apoptosis of HeLa cell was conformed by DNA ladder formation on agarose electrophoresis.Conclusion Serum containing Blaps femoralis Fischer-Waldheim has a marked inhibitory effect on proliferation of HeLa cell line and could induce apoptosis of HeLa cell.