论文部分内容阅读
目的观察沉默细胞凋亡抑制蛋白1(c IAP-1)基因表达对胰腺癌细胞HS766T放疗敏感性的影响。方法将HS766T细胞随机分为对照组、空载组和观察组,观察组和空载组分别转染沉默c IAP-1基因质粒c IAP-1-RNAi和阴性基因质粒NC-RNAi,对照组不转染处理,分别采用Real-time PCR法和Western blotting法检测细胞中的c IAP-1 mRNA及蛋白;取观察组c IAP-1稳定沉默HS766T细胞及其对照组HS766T细胞,用CCK-8法测算2、4、6、8 Gy X线照射24 h的细胞存活率,流式细胞仪测算6 Gy X线照射12、24、48 h的细胞凋亡率。结果观察组较空载组和对照组c IAP-1 mRNA及蛋白相对表达量降低(P均<0.05),空载组和对照组c IAP-1 mRNA及蛋白相对表达量无统计学差异。随着照射剂量增加,细胞存活率呈下降趋势;同等照射剂量下,观察组细胞存活率低于空载组和对照组(P均<0.05),空载组和对照组细胞存活率无统计学差异。随着照射时间的延长,各组细胞凋亡率呈现递增趋势;同一时点内,观察组细胞凋亡率高于空载组和对照组(P均<0.05),空载组和对照组细胞凋亡率无统计学差异。结论沉默c IAP-1基因表达,可抑制胰腺癌细胞存活、促进细胞凋亡,增强胰腺癌细胞的放疗敏感性。
Objective To investigate the effect of c IAP-1 gene silencing on the radiosensitivity of pancreatic cancer cell line HS766T. Methods HS766T cells were randomly divided into control group, no-load group and observation group. The control group and the no-load group were transfected with the c IAP-1 gene plasmid c IAP-1-RNAi and the negative gene plasmid NC-RNAi respectively. The expression of c IAP-1 mRNA and protein in the cells were detected by Real-time PCR and Western blotting respectively. Stably silenced the HS766T cells and its control HS766T cells by c IAP-1 in the observation group, CCK-8 method The cell viability was calculated at 2,4,6,8 Gy X ray irradiation for 24 h, and the apoptotic rate was measured by flow cytometry at 12, 24 and 48 h after 6 Gy X-ray irradiation. Results Compared with no-load group and control group, the relative expression of c-IAP-1 mRNA and protein in the observation group decreased (P <0.05). There was no significant difference in c IAP-1 mRNA and protein expression in the no-load group and the control group. With the increase of radiation dose, cell survival rate decreased. Under the same radiation dose, the survival rate of observation group was lower than that of no-load group and control group (all P <0.05), while no-cell survival rate difference. With the prolongation of irradiation time, the apoptotic rate of each group showed an increasing trend. In the same time point, the apoptosis rate of the observation group was higher than that of the control group and the no-load group (P <0.05) There was no significant difference in apoptosis rate. Conclusion The silencing of c IAP-1 gene expression can inhibit the survival of pancreatic cancer cells, promote apoptosis and enhance the radiosensitivity of pancreatic cancer cells.