MLN2238对宫颈癌HeLa细胞生长抑制和放射增敏及其机制探讨  

Radiation sensitization MLN2238 on human HeLa cells of cervical cancer

作　　者：葛杨杨[1,2] 谷庆[1] 马长升[1,3] 蔡晶[2] 王骁[1] 曹建平[1] 

机构地区：[1]苏州大学医学部放射医学与防护学院.江苏省放射医学与防护重点实验室,江苏苏州215123 [2]南通市肿瘤医院放疗科,江苏南通226361 [3]山东省肿瘤医院放疗科,山东济南250117

出　　处：《中华肿瘤防治杂志》2014年第4期246-251,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81172597;81102078);江苏省高校重大科研项目(11KJA310001)

摘　　要：目的:探讨蛋白酶体抑制剂MLN2238对宫颈癌HeLa细胞生长抑制和放射增敏作用及其机制。方法:采用CCK-8法检测MLN2238对HeLa细胞生长的抑制效应,克隆形成实验检测MLN2238辐射敏感性的变化,流式细胞术检测细胞凋亡率的变化,线粒体荧光探针显像检测线粒体活性变化,蛋白质印迹法测定细胞中蛋白表达情况。结果:MLN2238对HeLa细胞有明显的抑制作用,0.05μmol/L抑制率为3.95%,0.1μmol/L为14.89%,0.5μmol/L为29.37%,1μmol/L为38.95%,5μmol/L为54.44%,10μmol/L为70.52%,30μmol/L为81.76%,其效应呈剂量依赖性,F=1172.02,P<0.001。0.1μmol/L的MLN2238对HeLa细胞有放射增敏作用,其放射增敏比(sensitizing enhancement ratio,SER)为1.40。MLN2238联合X射线对HeLa细胞存在交互作用,对照组细胞凋亡分数为(2.64±0.07)%,单药组为(2.76±0.38)%,单照4Gy组为(9.50±0.14)%,单照8Gy组为(21.04±0.04)%,联合4Gy组为(11.12±0.19)%,联合8Gy组为(26.18±0.35)%,细胞凋亡率逐渐增加,差异有统计学意义,F=20.23,P=0.002 2。0.1μmol/L的MLN2238可影响细胞线粒体活性,导致线粒体膜损伤加重,增加细胞凋亡。MLN2238与X射线联合作用能增加凋亡相关蛋白Bax/Bcl-2的表达。结论:MLN2238对宫颈癌HeLa细胞有生长抑制和放射增敏作用,MLN2238对宫颈癌HeLa细胞可能通过线粒体途径介导的凋亡而增强其辐射敏感性。OBJECTIVE： To investigate the effect of proteasome inhibitor MLN2238 on human cervical cancer HeLa cell growth inhibition,it＇s radiosensitivity and its mechanism. METHODS： HeLa cell growth inhibitory effect was detected by CCK-8;colony formation assay was used to detect the radiosensitivity changes We used the flow cytometry to detect the rate of apoptosis;Mitochondrial fluorescent probe imaging was used to detect the changes in mitochondrial activity;Determination of protein expression in cells were detected by Western Blot method. RESULTS： The results showed that MLN2238 significantly inhibited HeLa cells,with the inhibition rate of 3.95% by 0.05 /μol/L, 14.89% by 0.1 μmol/L, 29.37% by 0.5 μmol/L,38.95% by 1 μmol/L,54.44% by 5 μmol/L,70.52% by 10 μmol/L,81.76% by 30 μmol/L,and its effect was dose-dependent, MLN2238（0. 1 μmol/L） showed the radiosensitizina effects on HeLa cells,and the sensitizing enhancement ratio （SER） was 1.40. MLN2238 interacts with X-ray in HeLa cells. The ap optosis rate of the control group was （2.64±0.07）%, the MLN2238 group was （2.76 ±.38）%, the 4 Gy radiation sin gle group was （9.50 ± 0.14）%, the 8 Gy radiation single group was （21.04 ± 0.04）%, the 4 Gy combination group was （11.12±0.19） % ,the 8 Gy combination group was （26.18±35） % ,the apoptosis rate was increased,the difference was statistically significant （F= 20.23 ,P±O. 002 2）. 0.1 /μol/L MLN2238 aggravated the menbrane injury while it can in crease apoptosis, the combination of X-rays and MLN2238 can increase the expression of protein Bcl-2 and Bax. CONCLU- SION： MLN2238 can inhibite HeLa cells growth and has radiosensitizing effect on HeLa cells, which may mediated through the mitochondrial pathway of apoptosis.

关 键 词：宫颈肿瘤 MLN2238 辐射增敏药 细胞凋亡 线粒体 

分 类 号：R737.33[医药卫生—肿瘤]