雷公藤红素逆转宫颈癌细胞顺铂耐药性实验研究  

An Active Component of Chinese Traditional Medicine Celastrol Reverses the Cisplatin-resistance of Cervical Cancer

作　　者：孙菊萍[1] 姜凯[2] 

机构地区：[1]浙江中医药大学附属第二医院检验科,杭州310005 [2]浙江省立同德医院检验科,杭州310012

出　　处：《浙江中西医结合杂志》2016年第7期622-626,共5页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的研究中药活性成分雷公藤红素逆转耐药宫颈癌细胞株Hela/R对顺铂的耐药性,并探讨其机制。方法采用顺铂梯度暴露法构建顺铂耐药宫颈癌细胞株Hela/R;采用MTT法检测雷公藤红素是否能增强顺铂对Hela/R的杀伤活性;Western blot试验检测常规Hela细胞及Hela/R细胞抗凋亡蛋白Bcl-2、Bcl-w及Bcl-xl表达水平;流式细胞术检测雷公藤红素联合顺铂对Hela/R细胞的凋亡诱导效应及对线粒体膜电位的影响。结果 1、2、4、8、16、32、64μmol/L顺铂对Hela及Hela/R细胞的细胞活力抑制率分别为:(18.6±1.5)%(Hela),(1.9±1.0)%(Hela/R);(23.9±2.4)%(Hela),(3.1±1.2)%(Hela/R);(47.8±3.9)%(Hela),(6.8±1.5)%(Hela/R);(63.4±5.4)%(Hela),(11.6±1.8)%(Hela/R);(72.7±5.9)%(Hela),(20.4±2.0)%(Hela/R),(85.7±6.7)%(Hela),(41.2±3.3)%(Hela/R);(91.8±7.9)%(Hela),(55.8±4.3)%(Hela/R),两组比较,P均<0.05。2μmol/L雷公藤红素对Hela/R细胞的细胞活力抑制率为(6.6±1.1)%;2μmol/L雷公藤红素分别加10、20、40μmol/L顺铂组Hela/R细胞的细胞活力抑制率优于单用10、20、40μmol/L顺铂组[(57.2±3.9)%比(12.4±1.2)%、(71.4±5.1)%比(27.8±1.9)%、(84.8±6.8)%比(45.2±3.1)%,P<0.05]。Hela/R细胞Bcl-w/β-actin表达水平高于Hela细胞[(70.9±4.9)比(22.1±1.3),P<0.05],Bcl-2/β-actin与Bcl-xl/β-actin表达水平差异无统计学意义(P>0.05)。与对照组比较,雷公藤红素可明显抑制Hela/R细胞Bcl-w/β-actin的表达[(29.3±2.6)比(68.9±4.5),P<0.05]。雷公藤红素增强顺铂对Hela/R细胞线粒体膜电位的损伤,促进细胞色素C的释放,而诱导Hela/R细胞发生凋亡。结论雷公藤红素体外能提高顺铂对耐药宫颈癌细胞的杀伤活性,其机制可能为通过抑制Bcl-w表达,提高促凋亡蛋白的活性,进而促进耐药宫颈癌细胞发生线粒体途径的凋亡。Objective To investigate the effect traditionl Chinese medicine celastrol on reversing the cisplatinresistance in cervical cancer cell line Hela/R and the underlying mechanism. Methods The cisplatin-resistant Hela/R cell line was established via stepped exposure to cisplatin. MTT assay was performed to explore whether the celastrol can enhance the cytotoxicity of cisplatin to Hela/R cells. The expression of Bcl-2, Bcl-w and Bcl-xl was evaluated by using western blot analysis. The mitochondrial membrane potential and apoptotic rate of Hela/R was determined by flow cytometry. Results The viability inhibition of Hela and Hela/R cells treated with cisplatin was as follows： 18.6%±1.5%（Hela） and 1.9%±1.0%（Hela/R） in 1μmol/L cisplatin group; 23.9%±2.4%（Hela） and3.1%±1.2%（Hela/R） in 2μmol/L cisplatin group; 47.8%±3.9%（Hela） and 6.8%±1.5%（Hela/R） in 4μmol/L cisplatin group; 63.4%±5.4%（Hela） and 11.6%±1.8%（Hela/R） in 8μmol/L cisplatin group; 72.7%±5.9%（Hela） and 20.4%±2.0%（Hela/R） in 16μmol/L cisplatin group; 85.7% ±6.7%（Hela） and 41.2% ±3.3%（Hela/R） in 32μmol/L cisplatin group; 91.8% ±7.9%（Hela） and 55.8% ±4.3%（Hela/R） in 64μmol/L cisplatin group; significant differences were found between two groups（all P〈0.05）. The viability inhibition of Hela/R cells treated with celastrol plus cisplatin was as follows： 6.6%±1.1% in 2μmol/L celastrol group; 12.4%±1.2% in 10μmol/L cisplatin group; 27.8%±1.9% in20μmol/L cisplatin group; 45.2%±3.1% in 40μmol/L cisplatin group; 57.2%±3.9% in 10μmol/L cisplatin＋ celastrol group; 71.45±5.1% in 20μmol/L cisplatin＋celastrol group; 84.8%±6.8% in 40μmol/L cisplatin＋ celastrol group; the viability inhibition was better in celastrol plus cisplatin groups than that in cisplatin single groups（all P〈0.05）. The level of Bcl-w/β-actin in Hela/R cells was higher than that in Hela cells（70.9±4.9 vs 22.1±1.3, P〈0.05）; the level of Bcl-2/β-actin and Bcl-xl/β-actin was not different be

关 键 词：耐药宫颈癌细胞株 顺铂 Hela/R 雷公藤红素 BCL-W 线粒体 凋亡 

分 类 号：R285[医药卫生—中药学]