顺铂诱导人宫颈癌Hela细胞凋亡后氧自由基水平的变化

Variation of oxygen free radicals after apoptosis induced by cisplatin in human cervical cancer Hela cells

目的探讨顺铂诱导人宫颈癌Hela细胞凋亡后氧自由基水平的变化特点。方法应用MTT比色法、流式细胞术、电子显微镜观察化疗前后Hela细胞活性、细胞周期,以及凋亡形态的改变;用化学比色法检测细胞凋亡前后细胞内丙二醛(MDA)、超氧化歧化酶(SOD)和谷胱甘肽(GSH)的含量。结果顺铂处理Hela细胞48 h的IC_(50)值为3 mg/L,当DDP浓度在3 mg/L以下时,对Hela细胞无明显毒性作用,超过此浓度时,其毒性呈剂量效应关系(P<0.001);与亲本细胞相比较,Hela+DDP细胞的G_2期细胞数增多,为(90.80±8.37)%,而G_1、S期的细胞数明显减少,分别为(30.84±8.02,5.30±0.60)%。扫描及透射电镜下可观察到典型的凋亡早期细胞形态学改变;当DDP作用48 h后,细胞内SOD的活性比对照组显著降低(P<0.05),细胞内的脂质过氧化产物MDA的含量与对照组相比显著升高(P<0.01),抗氧化剂GSH的含量较对照组明显降低(P<0.05)。结论经DDP可诱导Hela细胞的凋亡,其机制可能与细胞的氧化损伤有关。

Objective To study the variation of the intracellular oxygen free radicals after apoptosis induced by cisplatin in human cervical carcinoma cells. Methods Human cervical cancer cell line （Hela） were treated with different concentrations of DDP.The activation of the cell line, the changes in cell cycle, and the morphological changes of apoptosis cells were detected by MTY colori- metric assay, flow cytometry and electron microscopy, and the contents of intraeellular malondialdehyde （MDA） , superoxide dismutase （SOD） and giutathione （GSH） were detected by chemical colorimetric method. Results According to the MTT, the IC50 of DDP was 3 mg/L at the 48 h point, and DDP had no significant cytoxicity on Hela cell line when its concentration was less than 3 mg/L; contra- rily, the effects were dose - dependent when the concentration of DDP was higher than 3 mg/L （P 〈 0.001 ）. Compared with control group, the percentage of G2 phase ceils increased （90.80± 8.37） % in Hela ＋ DDP group, while the number of GI and S phase cells decreased significantly, the values were（30.84±8.02,5.30 ± 0.60） %, and （ P 〈 0.01 ）, respectively. The typical morphological changes of the early apoptotic cells could be observed by scanning and transmission electron microscopy. Compared with the control group, the activities of SOD and GSH decreased significantly（ P 〈 0.05） at the 48 hours point in the DDP group, and the concentration of MDA increased remarkably（P 〈0.01 ）. Conclusions DDP induces the death of Hela cells; the mechanism may be related to the oxidative damage in cells.