X线辐射后肺腺癌A549细胞上清外泌体跨膜蛋白88表达与细胞增殖的相关性  

Expression of exosome TMEM88 protein in supernatant of A549 cells after X irradiation and its relationship to cell proliferation

作　　者：王立威[1] 佟志敏 胡媛媛[3] 金鹏[3] 李宁[3] 陈玉丙[3] 

机构地区：[1]长春市人民医院,吉林长春130000 [2]吉林省申邦细胞工程研究所,吉林长春130000 [3]吉林大学第二医院,吉林长春130000

出　　处：《中国生物制品学杂志》2017年第10期1038-1041,1045,共5页Chinese Journal of Biologicals

基　　金：吉林省科技发展计划项目自然科学基金(20160101104JC)

摘　　要：目的探讨X线辐射对肺腺癌A549细胞增殖的影响,并观察其上清外泌体跨膜蛋白(transmembrane protein88,TMEM88)的表达情况。方法取对数生长期的肺腺癌A549细胞,分别用0、0.5、1、1.5、2 Gy(剂量率1.75 Gy/time,300 Mu/min)照射细胞,培养6和12 h后,CCK-8法检测不同辐射剂量对细胞增殖的影响;收集细胞上清,超速离心法提取外泌体蛋白,Western blot法分析外泌体TMEM88蛋白的表达情况。结果辐照后培养6 h,2 Gy组A450值明显小于0 Gy组及0.5 Gy组(P<0.05);其他各剂量组与0 Gy组比较,A450值呈下降趋势,但差异无统计学意义(P>0.05);辐照后培养12 h,与0 Gy组比较,各剂量A450值均降低,且呈剂量依赖性,除0.5 Gy组外,其他各剂量组差异均有统计学意义(P<0.05)。辐射后培养6 h,随辐射剂量增加,TMEM88蛋白相对表达水平呈下降趋势,2 Gy组与0 Gy组比较,差异有统计学意义(P<0.05);辐射后培养12 h,随辐射剂量增加,TMEM88蛋白相对表达水平呈逐渐下降趋势,且呈剂量依赖性,各组间差异均有统计学意义(P<0.05)。结论 X线辐射可抑制肺腺癌A549细胞的增殖及细胞上清外泌体TMEM88蛋白的表达。Objective To investigate the effect of X irradiation on proliferation of lung adenocarcinoma A549 cells and observe the expression of exosome transmembrane protein 88（TMEM88） in the supernatant. Methods A549 cells in logarithmic growth phase were irradiated with X-ray at dosages of 0, 0. 5, 1. 0, 1. 5, 2. 0 Gy（dose rate： 1. 75 Gy/time,300 Mu/min）, further cultured for 6 and 12 h, and determined for proliferation level by CCK-8 method. The supernatant was collected, from which the exosomes were extracted by ultracentrifugation. The expression of TMEM88 protein was determined by Western blot. Results The A450 value of cells 6 h after irradiation with 2 Gy X-ray was significantly lower than those with 0 and 0. 5 Gy（P〈0. 05）. The A450 value decreased with the increasing irradiation dose, while showed no significant difference in various dosage groups（P〈0. 05）. However, the A450 value 12 h after irradiation decreased in a dose-dependent pattern, and those in various dosage groups except 0. 5 Gy group showed significant difference with that in0 Gy group（P〈0. 05）. The relative expression level of TMEM88 protein 6 h after irradiation showed a decreasing tendency with the increasing irradiation dosage, which showed significant difference in 2 and 0 Gy groups（P〉0. 05）.However, the expression level 12 h after irradiation showed a gradually decreasing tendency with increasing irradiation dosage, which showed significant difference in various dose groups（P〈0. 05）. Conclusion X irradiation inhibited the proliferation of lung adenocarcinoma cell line A549 and the expression of exosome TMEM88 protein in the supernatant.

关 键 词：外泌体 跨膜蛋白88 辐射 肺癌细胞 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]