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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-ray radiation on the proliferation of lung adenocarcinoma A549 cells and to observe the expression of transmembrane protein 88 (TMEM88) in supernatant of the lung. Methods A549 lung adenocarcinoma cells in logarithmic growth phase were irradiated with 0,0.5,1,1.5,2 Gy (dose rate of 1.75 Gy / time, 300 Mu / min). After cultured for 6 and 12 h, CCK- 8 method to detect the effects of different doses of radiation on cell proliferation; collect the supernatant of the cells, extract the exosomes by ultracentrifugation, and analyze the expression of exosome TMEM88 by Western blot. Results The A450 value of 2 Gy group was significantly lower than that of 0 Gy group and 0.5 Gy group (P <0.05) at 6 h after irradiation. Compared with 0 Gy group, the A450 value of other dose groups showed a decreasing trend, but the difference was not statistically significant (P> 0.05). Compared with 0 Gy group, the A450 values of all dosages were decreased in a dose-dependent manner after irradiation for 12 h, except the 0.5 Gy group, the differences were statistically significant (P <0.05). Compared with 0 Gy group, the relative expression level of TMEM88 protein showed a decreasing trend at 6 h after irradiation (P <0.05). After 12 h of irradiation, the expression of TMEM88 protein decreased with the increase of radiation dose , TMEM88 protein relative expression levels showed a downward trend, and in a dose-dependent manner, the differences between the groups were statistically significant (P <0.05). Conclusion X-ray radiation can inhibit the proliferation of lung adenocarcinoma A549 cells and the expression of TMEM88 protein in the supernatant of the cells.