论文部分内容阅读
目的研究吉西他滨(Gem)诱导胰腺癌PANC-1细胞耐药与C-IAP2的关系。方法用间歇浓度递增法诱导胰腺癌细胞株PANC-1对吉西他滨耐药,将胰腺癌细胞株分为亲本株PANC-1组和耐药株PANC-1/Gem组。用四唑盐比色法检测细胞的半抑制浓度(IC_(50))值,用细胞计数法检测细胞倍增时间,用流式细胞术检测细胞周期,用免疫荧光法检测C-IAP2免疫荧光表达,用免疫印迹法检测C-IAP2表达变化。结果耐药株PANC-1/Gem组的IC_(50)值显著高于亲本株PANC-1组的IC_(50)值(P<0.05)。耐药株PANC-1/Gem组的倍增时间为32 h明显长于亲本株PANC-1组23 h(P<0.05)。2组细胞株在S期的比例差异无统计学意义(P>0.05)。与亲本株PANC-1组比较,耐药株PANC-1/Gem组表达出的C-IAP2荧光明显增强(P<0.05),C-IAP2蛋白表达水平显著升高(P<0.05)。结论间歇浓度递增法可诱导胰腺癌细胞株PANC-1对吉西他滨耐药,C-IAP2的上调表达在PANC-1/Gem耐药过程中起到一定作用。
Objective To investigate the relationship between gemcitabine-induced pancreatic cancer PANC-1 cell resistance and C-IAP2. Methods Pancreatic cancer cell line PANC-1 was induced to resist gemcitabine by intermittent concentration increasing method. Pancreatic cancer cell lines were divided into parental strain PANC-1 and resistant strain PANC-1/Gem. The semi-inhibitory concentration (IC_(50)) of the cells was measured with tetrazolium salt colorimetric method. The cell doubling time was measured by cell counting method. The cell cycle was detected by flow cytometry and the immunofluorescence assay was used to detect the immunofluorescence of C-IAP2. The expression of C-IAP2 was detected by immunoblotting. Results IC_(50) value of the resistant strain PANC-1/Gem was significantly higher than that of the parental strain PANC-1 (P<0.05). The doubling time of the drug-resistant PANC-1/Gem group was 32 h, which was significantly longer than that of the parental PANC-1 group (P<0.05). There was no significant difference in the proportion of S cell lines between the two groups (P>0.05). Compared with the parental strain PANC-1, the fluorescence of C-IAP2 expressed in the PANC-1/Gem group was significantly increased (P<0.05), and the expression of C-IAP2 protein was significantly increased (P<0.05). Conclusion The intermittent concentration increasing method can induce pancreatic cancer cell line PANC-1 to resist gemcitabine, and up-regulation of C-IAP2 plays a role in PANC-1/Gem resistance.