摘要
目的探讨唑来膦酸(Zoledronic acid,ZOL)对人直肠癌细胞株COLO320体外增殖和凋亡的影响,及其诱导凋亡可能的分子机制。方法采用CCK-8法检测不同浓度ZOL作用不同时间对COLO320细胞增殖的影响,并计算半数致死量(IC50)及增殖抑制率;采用流式细胞术(flow cytometry,FCM)分析ZOL对细胞凋亡的影响;采用Western blotting和实时定量PCR(Real-Time PCR)检测ZOL作用直肠癌COLO320细胞72 h后相关凋亡基因Bad、Bcl-2、Bax和Caspase-9的mRNA及蛋白表达水平。结果与对照组相比,ZOL处理组的直肠癌COLO320细胞增殖受到明显抑制,呈剂量和时间依赖性(P<0.05),24、48、72和96 h的IC50分别是209.4、103.6、74.1、65.6μmol/L;且ZOL对COLO320细胞增殖抑制作用呈时间和浓度依赖性。FCM示不同浓度ZOL处理直肠癌COLO320细胞72 h后的凋亡率较对照组均有不同程度升高,但20μmol/L浓度组与对照组之间差异无统计学意义(P>0.05),其余各组与对照组比较差异有统计学意义(P<0.05)。ZOL处理72 h后的COLO320细胞凋亡相关基因Bcl-2的表达水平下降,Bax、Bad、Caspase-9的表达升高。RT-PCR显示,Bcl-2 mRNA表达下调,Bax、Bad、Caspase-9 mRNA表达升高。结论 ZOL对人直肠癌COLO320细胞的增殖有抑制作用,对其凋亡有诱导作用。
Objective To investigate the effects and possible mechanisms of Zoledronic acid （ZOL） on the prolifera- tion and apoptosis of human rectal cancer COLO320 cell in vitro. Methods The proliferation of COLO320 cells exposed to different concentrations of ZOL for various periods was examined by the cell counting kit-8 （CCK-8） , the rate of inhi- bition and the half maximal inhibitory concentration （IC50） were calulated. Flow cytometry（FCM） showed ZOL induced apoptosis in COLO320 cells. The expressions of Bad, Bcl-2, Bax and Caspase-9 were measured by Western blotting and Real-Time PCR. Results Compared with the control group, ZOL treatment could remarkably inhibit the proliferation of rectal cancer COLO320 cells in a dose-and time-dependent manner （P 〈 0.05）. The values of IC50 were 209.4μ mol/L for 24 hours, 103.6 μmol/L for 48 hours, 74.1μmol/L for 72 hours, 65.6 μmol/L for 96 hours. FCM showed that the apoptosis rate of colorectal cancer cells COL0320 72 hours treated with different concentrations of ZOL were increased with different degrees compared with control group, but there was no significant difference between the 20 μmol/L con- centration group and the control group （P 〉 0.05） , there were statistically significant differences between control group and other groups （P 〈 0.05）. Meanwhile, Western blotting and Real-Time PCR results showed that the expression of Bcl-2 was down-regulated and expressions of Bad, Bax and Caspase-9 mRNA were up-regulated at a dose-dependent manner in COLO320 cells. Conclusion ZOL can inhibit the proliferation and induce apoptosis in rectal cancer CO- LO320 cells in vitro.
出处
《胃肠病学和肝病学杂志》
CAS
2016年第10期1171-1175,共5页
Chinese Journal of Gastroenterology and Hepatology
作者简介
田甜，硕士研究生，研究方向：消化道肿瘤的综合治疗。E—mail：814883815@qq．com
通讯作者：高超，教授，硕士生导师，主任医师，研究方向：消化道肿瘤的综合治疗。E-mail：gaochaoly@sina．com
