细胞分化剂对肿瘤细胞凋亡的诱导与耐药性的逆转

Induction of apoptosis and reversal of drug resistance in human tumor cell line KBV200 by cell differential agent Ⅱ

目的 探讨细胞分化剂 (CDA Ⅱ )诱导KBV2 0 0细胞调亡与逆转其耐药性的关系。方法 用四唑蓝比色法检测细胞存活率 ;用形态学 (光镜、透射电镜 )、生化学 [末端脱氧核苷酸转移标记法(Tunel法 ) ]、流式细胞术分析细胞凋亡。结果 CDA Ⅱ毒性作用很低。 1g/L和 2g/LCDA Ⅱ可使长春新碱 (VCR)对KBV2 0 0半数抑制浓度由 10 87.7nmol/L分别降低至 4 16.0nmol/L及 18.5nmol/L(P <0 .0 0 5及P <0 .0 0 1) ,2g/L逆转作用明显强于 1g/L(P <0 .0 5 ) ;CDA Ⅱ还与中药汉防已甲素有协同逆转作用。形态学及生化学均可观察到明显细胞凋亡 ,且与剂量相关。流式细胞术分析 ,CDA Ⅱ作用 96h后 ,1g/L、2g/L及 4g/L均可诱导细胞凋亡 ,凋亡指数分别为 (5 .4 3± 0 .4 3 ) %、(13 .5 0± 2 2 0 ) %和 (2 1.74± 0 .4 2 ) % ,各组间差异均有显著性 (P <0 .0 0 1,P <0 .0 5 ) ;2 0 0nmol/LVCR作用KBV2 0 0 96h后 ,未检出细胞凋亡 ;2 0 0nmol/LVCR和 1g/LCDA Ⅱ作用 96h后 ,凋亡指数为 (10 .88±0 .4 6) % ,较CDA Ⅱ (1g/L )单独作用 96h明显增高 (P <0 .0 0 1)。结论 CDA Ⅱ可逆转KBV2 0 0细胞耐药性与诱导其细胞凋亡 ,这种耐药性的逆转与CDA Ⅱ诱导细胞凋亡有关。

Objective To investigate the ability of cell differential agent Ⅱ (CDA Ⅱ) to reverse drug resistance and induce apoptosis in human tumor cell line KBV200. Methods Cell surviving fraction was detected by MTT assay; apoptosis in KBV200 was observed with light and electronic microscopys, and DNA end labeling method (Tunel).The apoptosis index (AI) was determined by flow cytometry (FCM). Results Cytotoxocity of CDA Ⅱ is low. However, 1 g/L and 2 g/L of CDA Ⅱ could increase cytotoxocity of VCR on KBV200 and IC 50 of VCR on KBV200 decreased from 1 087.7 nmol/L to 416.0 nmol/L and 18.5 nmol/L respectively ( P <0.005, and P <0.001). The reversal effect of 2 g/L of CDA Ⅱ was stronger than that of 1 g/L ( P <0.05); combination of CDA Ⅱ and TTD could increase the reversal effect. Apoptosis of KBV200 occurred after 96～120 hour treatment with CDA Ⅱ. The induction of apoptosis was related to the dosage of CDA Ⅱ. AI (%) of KBV200, analysed with FCM, was 5.43±0.43, 13.50±2.20 and 21.74±0.42 respectively, when KBV200 was treated by 1 g/L, 2 g/L and 4 g/L of CDA Ⅱ for 96 hours. There was significant difference between each group ( P <0 01, and P <0.05). Apoptosis of KBV200 was not induced by VCR (200 nmol/L) treatment for 96 hours alone. But AI (%) was 10.88±0.46, when KBV200 was treated with VCR (200 nmol/L) and CDA Ⅱ (1 g/L) for 96 hours. AI was significantly higher than treated with CDA Ⅱ alone ( P <0.005). Conclusion CDA Ⅱ could induce apoptosis and reverse durg resistance of KBV200. The reversal effect was related to the induction of apoptosis by CDA Ⅱ.