论文部分内容阅读
采用大剂量冲击、间歇诱导法获得人肝癌顺铂(cisplatin,CDDP)多药耐药细胞系SK-Hep1/CDDP。利用线粒体通透转运孔道(mitochondrial permeability transition pore,mPTP)开放剂苍术苷(atractyloside,ATR)和抑制剂环孢素A(cyclosporin,CsA)分别干预SK-Hep1和SK-Hep1/CDDP细胞;免疫印迹法检测多药耐药基因Mdr-1和Bax表达水平;采用Annexin V/PI双标记法检测细胞凋亡率;采用荧光探针JC-1检测线粒体膜电位△Ψm的变化。探讨调控线粒体mPTP对人肝癌耐药细胞SK-Hep1/CDDP多药耐药的影响。结果提示,ATR可促进mPTP开放,加速△Ψm下降,同时降低Bax活性,增加SK-Hep1/CDDP细胞凋亡;CsA抑制mPTP开放,可减轻并延迟线粒体膜电位下降,使多药耐药细胞SK-Hep1/CDDP对CDDP诱导凋亡的耐受能力提高,同时增加Bax活性。但mPTP活性的变化对细胞Mdr-1蛋白表达水平无影响。mPTP的激活可能成为增加肿瘤细胞对化疗药物敏感性及逆转肿瘤细胞多药耐药的新方法。
The multi-drug resistant cell line SK-Hep1 / CDDP of human hepatocarcinoma cisplatin (CDDP) was obtained by high-dose impact and intermittent induction. The mitochondria permeability transition pore (mPTP) openers atractyloside (ATR) and cyclosporin A (CsA) were used to interfere SK-Hep1 and SK-Hep1 / CDDP cells respectively. The expression of multidrug resistance gene Mdr-1 and Bax was detected by flow cytometry. The apoptosis rate was detected by Annexin V / PI double labeling method. The change of mitochondrial membrane potential △ Ψm was detected by fluorescent probe JC-1. To investigate the effects of mitochondrial mPTP on multidrug resistance of human hepatocellular carcinoma cell line SK-Hep1 / CDDP. The results suggest that ATR can promote the opening of mPTP, accelerate the decrease of △ Ψm, decrease the activity of Bax and increase the apoptosis of SK-Hep1 / CDDP cells. CsA can inhibit the opening of mPTP and reduce and delay the decrease of mitochondrial membrane potential, -Hep1 / CDDP enhances tolerance of CDDP-induced apoptosis while increasing Bax activity. However, changes in mPTP activity had no effect on the expression of Mdr-1 protein. Activation of mPTP may become a new method to increase the sensitivity of tumor cells to chemotherapeutic drugs and reverse the multidrug resistance of tumor cells.