顺铂诱导人宫颈癌细胞系P27表达及其超微弱发光测定的意义  

Cisplatin DDP induced P27 expression and ultra weak luminescence detection in human cervical carcinoma cell line

作　　者：杨海宁[1] 惠延平[1] 辛晓燕[2] 黄艳红[2] 王波 

机构地区：[1]第四军医大学西京医院病理科,陕西西安710032 [2]第四军医大学西京医院妇产科,陕西西安710032 [3]军事预防医学系流行病医学教研室,陕西西安710032

出　　处：《现代生物医学进展》2009年第3期414-417,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金(60471035)

摘　　要：目的:探讨化疗药物对肿瘤增殖活性的影响。方法:选择人宫颈癌细胞系Hela分为两组,分别采用MTT比色法分析测定顺铂处理Hela细胞的浓度;免疫组化SP法分别检测Hela细胞中P27蛋白表达;流式细胞仪分析加药前后细胞周期变化及凋亡情况;用IFFM-D型流动式化学发光仪检测细胞的超弱发光强度。结果:顺铂处理Hela细胞48 h的IC50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);流式细胞仪分析细胞周期可见与Hela细胞相比较,Hela+DDP细胞的G2期细胞数增多,而G1、S期的细胞数明显减少(P<0.01);从细胞凋亡检测显示Hela与Hela+DDP相比,细胞凋亡率在不同时间点明显升高,在24 h、48 h、72 h结果分别为(11.4±5.8、21.8±7.9、32.5±11.6)%。免疫组化结果显示Hela+DDP与Hela细胞相比细胞膜上P27蛋白高表达;在10-4mol/L鲁米诺及0.3%的双氧水(H_2O_2)条件下Hela细胞超弱发光强度高于用Hela+DDP细胞(P<0.001)。结论:超弱发光能够快速、准确、有效地反映肿瘤细胞氧化代谢特点和增殖活动,也可用于筛选敏感的化疗药物。Objective： To investigate the effect of chemotherapeutics on proliferation activity of tumor. Methods： Human cervical carcinoma Hela cells were divided into two groups： cells treated with or without DDP. MTT chromatometry method was used to evaluate the concentration of DDP for Hela cells. Immunochemistry method was used to detect the P27 expression. Cell flow cytometry was used to detect the change of cell cycle and apoptosis. IFFM-D chemoluminescence machine was used to detect the ultra weak luminescence intensity. Results： The IC50 of Hela cells treated with cis-DDP for 48h was 3mg/L. When cis-DDP concentration was lower than 3 mg/L, the cell toxicity showed dose-effect relation （P〈0.001）. Cell flow cytometry results showed that Hela cells treated with DDP appeared more G2 phase and less G1 and S phase than control cell （P〈0.01）; Compared with Hela treated with DDP, the apoptosis rates of Hela cells were increased signifcantly at 24 h, 48 h and 72 h （11.4± 5.8, 21.8± 7.9, 32.5± 11.6） %. P27 expression was increased in Hela cells treated with DDP. Luminous intensity of Hela cells treated with 10- 4 mol/L luminol and 0.3 % H2O2 were significantly higher than that of Hela cells treated with DDP （P〈0.001）. Conclusions： Ultra weak luminescence can reflect the metabolism and proliferation of tumor cells, which can be used for screening sensitive chemotherapy drugs.

关 键 词：宫颈肿瘤 超微弱发光 凋亡 P27表达 抗肿瘤药 

分 类 号：R737.3[医药卫生—肿瘤]