目的研究卵巢癌特异性结合肽(ovarian cancer specific targeting peptide,OSTP)与顺铂(cis-dichlorodiamineplatinum,DDP)偶联物(OSTP-DDP)对卵巢癌A2780细胞的靶向抑制作用。方法化学合成OSTP-DDP,体外培养卵巢癌A2780细胞,采用CCK-8...目的研究卵巢癌特异性结合肽(ovarian cancer specific targeting peptide,OSTP)与顺铂(cis-dichlorodiamineplatinum,DDP)偶联物(OSTP-DDP)对卵巢癌A2780细胞的靶向抑制作用。方法化学合成OSTP-DDP,体外培养卵巢癌A2780细胞,采用CCK-8法分别检测OSTP-DDP和DDP对卵巢癌A2780细胞的生长抑制作用;通过Annexin V-FITC凋亡试剂盒分别检测OSTP-DDP和DDP对卵巢癌A2780细胞的周期和凋亡作用。结果通过质谱分析和高效液相色谱(HPLC)分析,提示成功合成OSTP-DDP。CCK-8法检测显示,OSTP-DDP与DDP分别以不同浓度(10、20、40、80、160、320μmol·L^(-1))处理卵巢癌A2780细胞24、48、72 h后,均可对该细胞的生长起到抑制作用,且呈时间、浓度依赖性。且OSTP-DDP的作用强于DDP(P<0.05),提示OSTP-DDP具有靶向抑制细胞生长作用。流式细胞术检测结果显示,经OSTP-DDP和DDP处理后,细胞周期均被阻滞于G_1期,作用72 h后,OSTP-DDP对卵巢癌A2780细胞的周期抑制作用强于DDP,差异具有统计学意义(P<0.05)。作用24、48、72 h后,OSTP-DDP对卵巢癌A2780细胞凋亡的作用强于DDP(P<0.01)差异具有统计学意义,提示OSTP-DDP具有较强的靶向诱导细胞凋亡作用。结论 OSTP-DDP对卵巢癌A2780细胞的生长具有靶向抑制作用,OSTP作为化疗药物靶向载体治疗卵巢癌有良好的应用前景。展开更多
To investigate the relationship between MDR1 and MDR3 gene and drug resistance to cisplatin of ovarian cancer cells. Two siRNAs （MDRI, MDR3） which specifically targeted MDRI and MDR3 genes were transfered into A2780...To investigate the relationship between MDR1 and MDR3 gene and drug resistance to cisplatin of ovarian cancer cells. Two siRNAs （MDRI, MDR3） which specifically targeted MDRI and MDR3 genes were transfered into A2780/DDP cells. Then double staining with Annexin- V-FITC/PI was used to detect cell apoptosis by the flow cytometry （FCM）. A2780/DDP cell viability was determined by MTT. MDR1 and MDR3 mRNA were assessed by RT-PCR. Caspase-3 protein was detected by Western blotting. Transfection of MDRI and MDR3 siRNA into A2780/DDP cells failed to reverse the drug-resistance of A2780/DDP cells to cisplatin （P〉0.05）. No significant difference in the apoptosis efficiency was observed between the MDR1 and MDR3 siRNA, pSuppressor- Neo vector transfection cells and untreated cells （P〉0.05）. In the presence of cisplatin of different concentrations, the viability of A2780/DDP cells was not significantly decreased after the transfection No changes in MDR1 and MDR3 mRNA were found in MDRI and MDR3 siRNA-transfected A2780/DDP cells. As compared with pSuppressorNeo and untreated groups, no significant difference existed in the expression of MDR1 and MDR3 mRNA （P〉0.05）. The expression of caspase-3 protein in MDR1 and MDR3 siRNA transfected A2780/DDP cells was not significantly increased. It is concluded that multidrug resistance induced by cisplatin in ovarian carcinoma cell lines is not due to overexpression of MDR1 and MDR3 gene. The drug resistance of ovarian carcinoma cells to cisplatin is not mediated by P-glycoprotein.展开更多
文摘目的研究卵巢癌特异性结合肽(ovarian cancer specific targeting peptide,OSTP)与顺铂(cis-dichlorodiamineplatinum,DDP)偶联物(OSTP-DDP)对卵巢癌A2780细胞的靶向抑制作用。方法化学合成OSTP-DDP,体外培养卵巢癌A2780细胞,采用CCK-8法分别检测OSTP-DDP和DDP对卵巢癌A2780细胞的生长抑制作用;通过Annexin V-FITC凋亡试剂盒分别检测OSTP-DDP和DDP对卵巢癌A2780细胞的周期和凋亡作用。结果通过质谱分析和高效液相色谱(HPLC)分析,提示成功合成OSTP-DDP。CCK-8法检测显示,OSTP-DDP与DDP分别以不同浓度(10、20、40、80、160、320μmol·L^(-1))处理卵巢癌A2780细胞24、48、72 h后,均可对该细胞的生长起到抑制作用,且呈时间、浓度依赖性。且OSTP-DDP的作用强于DDP(P<0.05),提示OSTP-DDP具有靶向抑制细胞生长作用。流式细胞术检测结果显示,经OSTP-DDP和DDP处理后,细胞周期均被阻滞于G_1期,作用72 h后,OSTP-DDP对卵巢癌A2780细胞的周期抑制作用强于DDP,差异具有统计学意义(P<0.05)。作用24、48、72 h后,OSTP-DDP对卵巢癌A2780细胞凋亡的作用强于DDP(P<0.01)差异具有统计学意义,提示OSTP-DDP具有较强的靶向诱导细胞凋亡作用。结论 OSTP-DDP对卵巢癌A2780细胞的生长具有靶向抑制作用,OSTP作为化疗药物靶向载体治疗卵巢癌有良好的应用前景。
文摘To investigate the relationship between MDR1 and MDR3 gene and drug resistance to cisplatin of ovarian cancer cells. Two siRNAs （MDRI, MDR3） which specifically targeted MDRI and MDR3 genes were transfered into A2780/DDP cells. Then double staining with Annexin- V-FITC/PI was used to detect cell apoptosis by the flow cytometry （FCM）. A2780/DDP cell viability was determined by MTT. MDR1 and MDR3 mRNA were assessed by RT-PCR. Caspase-3 protein was detected by Western blotting. Transfection of MDRI and MDR3 siRNA into A2780/DDP cells failed to reverse the drug-resistance of A2780/DDP cells to cisplatin （P〉0.05）. No significant difference in the apoptosis efficiency was observed between the MDR1 and MDR3 siRNA, pSuppressor- Neo vector transfection cells and untreated cells （P〉0.05）. In the presence of cisplatin of different concentrations, the viability of A2780/DDP cells was not significantly decreased after the transfection No changes in MDR1 and MDR3 mRNA were found in MDRI and MDR3 siRNA-transfected A2780/DDP cells. As compared with pSuppressorNeo and untreated groups, no significant difference existed in the expression of MDR1 and MDR3 mRNA （P〉0.05）. The expression of caspase-3 protein in MDR1 and MDR3 siRNA transfected A2780/DDP cells was not significantly increased. It is concluded that multidrug resistance induced by cisplatin in ovarian carcinoma cell lines is not due to overexpression of MDR1 and MDR3 gene. The drug resistance of ovarian carcinoma cells to cisplatin is not mediated by P-glycoprotein.
