复方苦参注射液对宫颈癌细胞体外放射增敏作用研究

Radiosensitization effect of Compound Kushen Injection on cervical cancer Hela cell cultured in vitro

目的研究复方苦参注射液对人宫颈癌Hela细胞株的体外放射增敏作用并初步探讨其作用机制。方法克隆形成法检测放射增敏作用及给药时序、浓度对细胞存活率的影响;DAPI染色法镜下观察细胞凋亡情况;流式细胞法检测细胞周期分布情况;Western blot法检测bcl-2、bax蛋白表达情况。结果复方苦参注射液与射线单独或联合作用后细胞存活率均较单纯放射组明显降低(P均<0.05),不同浓度、不同时序处理后细胞存活率比较差异有统计学意义,先照射后给药效果优于先给药后照射(P<0.05);复方苦参注射液与射线均能使Hela细胞阻滞于G2/M期,联用阻滞效果更加显著(P<0.05);复方苦参注射液与射线均能破坏细胞核,联用后细胞核皱缩、碎裂更加显著;复方苦参注射液与射线单独及联用均能上调bax蛋白表达(P均<0.05),下调bcl-2蛋白表达(P均<0.05)。结论复方苦参注射液可通过阻滞宫颈癌Hela细胞周期于G2/M期,下调bcl-2/bax的比值来诱导细胞发生凋亡并增强其对放射线的敏感性。

Objective It is to investigate the radiosensitization effect of Compound Kushen Injection on cervical cancer He -la cell and to primarily investigate its mechanism .Methods Clone forming assay was used to observe the radiosensitizing effect;Flow cytometry method was used to detect Hela cell cycle distribution; Apoptotic changes was measured by Fluores-cence microscope and DAPI staining; The expression of bcl -2 and bax protein was detected by Western blotting .Results Compared with radio alone,the cell survival rates was reduced by combination with Compound Kushen Injection （P〈0.05）. The cell survival rates were also depended on drug by various schedules and drug concentration （P〈0.05）.Compound Kush-en Injection alone or combined with radiation could arrest cell cycle in G 2/M phase（P〈0.01）,induce cell apoptosis, and the effects of combination were more significant （P〈0.05）.Compound Kushen Injection and radiation alone or combination all could damage the cell nucleus , the nudeus shrinkage and fragmentation was more significant in combination group （ P 〈0.05 ） .The levels of protein of bcl -2 was decreased and bax was increased by Compound Kushen Injection and radiation a -lone or combination （P〈0.05）.Conclusion Compound Kushen Injection has radiosensitization effect on Hela cell cultured in vitro ,which may be related to arrest in G 2/M phase, induce cell apoptosis and down-regulating the ratio of bcl-2/bax.