摘要
目的:探讨锌对前列腺癌细胞PC-3M凋亡的影响。方法:分别选用不同浓度ZnSO4·7H2O(终浓度分别为6·25、15、25、50、100、150、200、250及300μmol·L-1)作用于PC-3M细胞24h,用MTT法筛选对PC-3M细胞增殖抑制的有效浓度,利用吖啶橙/溴化乙锭染色、DNAladder及流式细胞术检测PC-3M细胞凋亡情况。结果:MTT显示,锌的浓度在250μmol·L-1时,PC-3M细胞存活率显著低于对照组(P<0·01)。Zn2+浓度达250μmol·L-1时,相差显微镜下细胞形态变化明显,细胞变长,并伸出较长的突起,细胞分裂相明显减少,细胞数量较对照组明显减少。吖啶橙/溴化乙锭染色显示,对照组细胞形态为规则的圆形,细胞膜光滑无皱缩和发泡,呈现均匀的绿色,偶可见橙色细胞(自然凋亡细胞);Zn2+(250μmol·L-1)作用后,相差显微镜下可观察到较多早期凋亡细胞,细胞多为圆形,但细胞膜较对照组粗糙,细胞呈绿色,细胞核中有鲜绿色的斑点;也可观察到较多的晚期凋亡细胞,细胞形状不规则,细胞膜粗糙,有较多突起,被吖啶橙染成橙色,细胞中有鲜亮的橙色斑点。DNAladder可见梯形条带出现;流式细胞术显示亚二倍体峰的出现,细胞凋亡率为13·3%。上述各项指标均显示PC-3M细胞的凋亡水平明显高于对照组(P<0·05)。结论:高Zn2+可以抑制前列腺癌细胞生长并诱导前列腺癌细胞凋亡。
Objective To investigate the effects of zinc on apoptosis of prostate cancer cells （PC-3M）. Methods The cell proliferation was determined by MTT assay treated with different concentrations of ZnSO4·7H2O （end concentrations were 6.25, 15, 25, 50, 100, 150, 200, 250, and 300μmol·L^-1, respectively）, the cell morphological changes after exposed to zinc for 24 h was observed under phase-contrast microscope, the apoptosis was detected by AO/EB assay, the cell cycle and apoptotic peak were detected by flow cytometry, DNA fragmentation was observed by agarose gel electrophoresis （AGE）. Results PC-3M cells proliferation rate was obviously changed after exposed to different levels of zinc for 24 h. Zinc （250μmol·L^-1） inhibited proliferation of PC-3M cells （P〈0. 01）. Compared with control cells, the changes of morphological characteristics of PC-3M cells were found, including shrank and floated, decreased refraction in the cells exposed to zinc （250 μmol·L^-1） for 24 h. The confocal microscopy showed the changes of cells through AO/EB staining, The cellular membrane was integrity but vacuolization of the cytoplasm, chromatin was flavo-green, nuclear condensed, nuclear side collection, caryolysis, nuclear fragmented. The flow cytometry showed the hypodip-loid peak in G0/G1 phase after exposed to zinc （250μmol·L^-1） for 24 h. It had visible DNA fragmentation by AGE.Conclusion Zincwith high concentrationcan inhibit human prostate cancer cell growth and induce apoptosis of human prostate cancer cells.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2006年第5期808-811,共4页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅资助课题(20030710)
