羟基喜树碱对前列腺癌PC-3细胞凋亡作用的研究

Hydroxycamptothecin Promotes the Apoptosis of Prostate Cancer Cell Line PC-3

目的:观察羟基喜树碱(HCPT)对前列腺癌PC-3细胞凋亡的影响,初步探讨其作用机制。方法:四甲基偶氮唑盐(MTT)检测不同浓度(1×10-1、1×10-2、1×10-3、1×10-4mg/ml)、不同作用时间(12、24、48h)下HCPT对PC-3细胞增殖的影响;吖啶橙/溴化乙锭(AO/EB)染色观察细胞凋亡情况;琼脂糖凝胶电泳观察凋亡细胞特征性DNA条带;流式细胞术测定HCPT诱导PC-3细胞凋亡率。结果:HCPT明显抑制PC-3细胞生长,呈时间、剂量依赖性,12、24、48h的IC50值分别为:6.50×10-2、2.35×10-2、5.31×10-3mg/ml;荧光显微镜下观察到经AO/EB染色的典型凋亡细胞;琼脂糖凝胶电泳可见凋亡细胞DNA呈规律的梯状条带;流式细胞术显示PC-3细胞凋亡率随HCPT浓度的增加而升高,浓度为1×10-3mg/ml时凋亡率达到最高峰(35.76%)。结论:HCPT可通过诱导凋亡较为明显地抑制PC-3细胞生长,但其作用机制目前尚需进一步研究。

Objective： To observe the effects of hydroxycamptothecin （HCPT） on the apoptosis of prostate cancer cell line PC-3 and to explore the possible mechanism. Methods： The influence of different concentrations （ 1 × 10^-1 , 1 × 10^-2, 1 ×10^-3 , 1 × 10^-4 mg/ml）of HCPT on PC-3 cell proliferation at different time （ 12, 24, 48 h）was determined by tetrazolium （MTF） assay. The morphologic changes of the apoptotic cells were observed by acridine orange / ethidium bromide dyeing. The DNA of the apoptotic cells was analyzed with agarose gel electrophoresis. The apoptosis rate of HCPT on prostate cancer cells was analyzed by flow cytometry （FCM）. Results： The growth of PC-3 was inhibited by HCPT in a time- and dose- dependent manner. The values of IC50 were 6.50× 10^-2 mg/ ml （ 12 h）, 2.35 × 10^-2 mg/ml （24 h） and 5.31 × 10^-3mg/ml （48 h） respectively. The typical apoptotic cells under the fluores- cence microscope showed budding phenomena and apoptotic bodies. And the DNA ladder was observed in ultraviolet light. FCM analysis showed that the apotosis rate of PC-3 cells increased with the increasing dose of HCPT, which reached the peak （35.76%） at 1 × 10^-3 mg/ml. Conclusion： HCPT could suppress PC-3 cell proliferation significantly by inducing the apoptosis of PC-3 cells. However, the mechanism is yet to be further studied.