纳秒脉冲诱导肿瘤细胞缝隙连接通讯恢复的实验研究

Experimental Study of Tumor Cell Gap Junctional Intercellular Communication Recovery with Nanosecond Pulsed Electric Fields

将纳秒脉冲作用于人肝癌细胞(Hep-G2),通过荧光漂白恢复(FRAP)技术,检测荧光萃灭后肿瘤细胞的荧光强度变化情况,以研究纳秒脉冲对肿瘤细胞缝隙连接通讯(GJIC)功能变化的影响。经纳秒脉冲处理后,与对照组相比,处理组荧光恢复率显著增加,且随着时间的推移,所有处理组的荧光恢复率都达到对照组的3倍以上。此外,处理组肿瘤细胞的荧光漂白恢复率随脉冲个数的增加而升高。实验结果表明,纳秒脉冲促进了肿瘤细胞缝隙连接通讯功能的恢复,且在固定的脉冲宽度和幅值下,GJIC对纳秒脉冲作用的响应程度与施加的脉冲个数呈正相关。研究结果不仅为纳秒脉冲用于GJIC障碍性疾病的治疗提供了一条全新的途径,而且也深化了纳秒脉冲诱导肿瘤细胞凋亡机制的研究。

In order to study the effect of nanosecond pulsed electric fields （nsPEF） on gap junctional intercellular communication （GJIC） of tumor cell, nsPEF were put on Hep-G2 ceils, then changes of fluorescence intensity after cancellation were tested by fluorescence recovery after photobleaching （FRAP） technology. Significant changes of fluorescence intensity after cancellation were observed in treated groups compared with that in control group, and the rate of fluorescence in all treated group was at least as three times as that in control group with time evolution. In addition, the rate of fluorescence recovery after photobleaching went up with the increase of pulse number. It was revealed that the recovery of GJIC of tumor cells could be triggered by nsPEF, and the response of GJIC to nsPEF was positive correlation with pulse number. Our research not only provides a complete way to treat GJIC-handicap diseases, but also deepens the mechanism studies of tumor cell apoptosis induced by nsPEF.