2.45GHz微波非热效应诱导人宫颈癌细胞凋亡及其机制

目的 观察2.45 GHz微波非热效应对人宫颈癌HeLa细胞凋亡的影响,探讨不同照射功率与凋亡诱导效应之间是否存在量效关系及其机制.方法 HeLa细胞随机分为对照组、假照射组和低、中、高(10、15和20 mW/cm²)3个功率照射组,2.45 GHz微波照射10 min.MTT法检测HeLa细胞存活率;激光共聚焦显微镜观察各组HeLa细胞核形态变化,并统计异常细胞核比率;流式细胞仪检测细胞周期并分析sub-G₁期细胞所占比例;免疫印迹法观察Bax、Bcl-2和Caspase-3凋亡相关蛋白表达变化并以β-肌动蛋白为内参比较各组蛋白表达灰度变化.结果 MTT结果显示,对照组与假照射组、低功率照射组之间细胞存活率无明显变化,中、高功率照射组HeLa细胞存活率较假照射组明显降低(t=-16.47、-14.23,P<0.05);激光共聚焦观察经Hoechst单染的细胞核显示,低、中、高功率照射组异常细胞核比率与假照射组相比明显增高(t=9.37、11.25、8.47,P<0.05),且高功率组细胞核异常比率高于低、中功率组(t=15.32、10.25,P<0.05);流式细胞术结果显示低、中、高功率照射组sub-G₁期细胞所占比例均比假照射组升高(t=15.24、22.31、10.69,P<0.05);免疫组织化学结果显示Bcl-2的表达随着微波照射功率的增大而减弱,Bax和Caspase-3的表达随着照射功率的增强而加强.结论 2.45 GHz微波可诱导HeLa细胞凋亡,且与其功率密度增加呈正相关.

The mechanism of non-thermal effect of 2.45 GHz microwave on HeLa cell apoptosis

Objective To investigate the effects of 2.45 GHz microwave on the apoptosis of HeLa cells and explore the underlying mechanism.Methods HeLa cells were divided into five groups randomly: control group, sham-irradiation group, and three irradiation groups exposed with 10,15 and 20 mW/cm² 2.45 GHz microwave for 10 min. Cell survival was tested by MTT assay, cell nucleus fashion was observed by a laser scanning confocal microscope, cell cycle distribution was measured by flow cytometry, and cell protein expressions were detected by Western blot. Results The cell survival in control group, sham-irradiation group and low dose irradiation group had no difference but it became lower in the middle and high dose irradiated group(t=-16.47, -14.23, P<0.05). The abnormal rates of nuclear fashion in the low,middle and high dose irradiation groups were obviously higher compared with control group(t=9.37, 11.25, 8.47, P<0.05), while high dose irradiation group was higher than the low and middle dose group (t=15.32, 10.25, P<0.05). The rate of HeLa cells in sub-G₁ in the irradiation groups were higher than that in control group(t=15.24, 22.31, 10.69, P<0.05). The results of immunohistochemistry showed that the expression of Bax and Caspase-3 in irradiation groups was significantly increased, and the expression of Bcl-2 was decreased. Conclusions 2.45 GHz microwave can induce HeLa cells apoptosis with a positive correlation with microwave dose.