陵水暗罗提取物暗罗素抗人肾癌细胞效应研究

Antitumor activity of Polyalthia memoralis extract zinc pyrithione in human kidney cancer cells lines

目的:研究陵水暗罗提取物暗罗素(ZPT)抑制肾癌细胞株的效应。方法:MTS法检测ZPT联合ZnCl_(2)对肾癌细胞786-O和769-P的增殖抑制效应,并计算两株细胞ZPT的IC_(50),同时检测ZPT联合ZnCl_(2)对两株肾癌细胞增殖抑制的浓度效应和时间效应;克隆形成实验检测ZPT联合ZnCl_(2)对两株细胞克隆形成的影响,流式细胞仪检测ZPT联合ZnCl_(2)对两株细胞凋亡的影响。结果:ZPT联合50μmol/L ZnCl_(2)可以抑制肾癌细胞株786-O和769-P的增殖活性,其效应呈剂量依赖性和时间依赖性,联合50μmol/L ZnCl_(2)处理72 h后,IC_(50)值分别为501.8 nmol/L和694.7 nmol/L。固定ZnCl_(2)浓度为50μmol/L,随着ZPT浓度增加,两株细胞的成活率逐渐下降,ZPT剂量为3 200nmol/L时,786-O和769-P细胞存活率分别为20.23%和24.75%。500 nmol/L和700 nmol/L ZPT联合50μmol/L ZnCl_(2)处理786-O和769-P细胞随处理时间增加细胞增殖抑制效应增强,处理时间为72 h后786-O和769-P细胞存活率分别为43.55%和38.16%。克隆形成实验显示500 nmol/L和700 nmol/L ZPT联合50μmol/L ZnCl_(2)处理786-O和769-P细胞10 d后,两株细胞的克隆形成率分别为对照组(DMSO)的40.24%和45.24%。流式细胞仪检测细胞凋亡水平显示:500 nmol/L ZPT联合50μmol/L ZnCl_(2)处理786-O 12 h后凋亡明显增加(DMSO vs ZPT+ZnCl_(2)组:1.67%vs 36.04%)。结论:暗罗素联合锌离子可以抑制肾癌细胞株增殖,其效应呈剂量和时间依赖性;同时抑制两株细胞的克隆形成率,增加肾癌细胞的凋亡。

Objectives: To investigate the antitumor activity of Polyalthia memoralis extract zinc pyrithione(ZPT)in human kidney cancer cells lines. Methods: Cell viability of ZPT plus 50 μmol/L ZnCl_(2) on786-O and 769-P cell lines was detected with MTS method. The average inhibitory concentration 50 values(IC_(50) values) were determined after 48-hour treatment. The dose-course and time-course effect of ZPT plus 50 μmol/L ZnCl_(2) in two cells lines was measured with MTS methods. The colony formation ratio and apoptosis of ZPT plus 50 μmol/L ZnCl_(2) on two cells lines were determined with colony formation assay and flow cytometry. Results: ZPT plus 50 μmol/L ZnCl_(2) could inhibit the proliferation of 786-O and 769-P cell lines with the IC_(50) values of 501. 8 nmol/L and 694. 7 nmol/L, respectively, after 48-hour treatment. The proliferation inhibition effect was time-and dose-dependent. The inhibition rates of 3. 2 μmol/L ZPT plus 50 μmol/L ZnCl_(2) on 786-O and 769-P cells respectively were20. 23% and 24. 75% respectively after 48-hour treatment. While the rates of 500 nmol/L ZPT for786-O and 700 nmol/L for 769-O in the presence of 50 μmol/L ZnCl_(2) were 43. 55% and 38. 16% respectively after 72-hour treatment. ZPT plus 50 μmol/L ZnCl_(2) inhibited the colony formation in two cell lines(DMSO vs ZnCl_(2) vs ZPT+ZnCl_(2): 100. 00% vs 98. 40% vs 40. 24% for 786-O cells, P<0. 01;100. 00% vs 104. 37% vs 45. 24% for 769-P cells, P<0. 01). The apoptosis ratio was enhanced in 786-O cell line after 12-hour treatment of 500 nmol/L ZPT+50 μmol/L ZnCl_(2)(Apoptosis ratio: 1. 31%, 1. 85%, 3. 69%, and 25. 15%, respectively in the groups of DMSO, ZnCl_(2), ZPT,and ZPT+Zn_(2), respectively, P<0. 01). Conclusion: ZPT plus 50 μmol/L ZnCl_(2) can inhibit the proliferation of 786-O and 769-P cell lines, and ZPT presents the antitumor activity in the human kidney cancer cell lines.