摘要
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 observe the effect of silencing cellular inhibitor of apoptosis protein 1（c-IAP1） on the sensitivity of pancreatic cancer HS766 T cells to radiotherapy. Methods HS766 T cells were randomly divided into the control group,empty vector group,and observation group. The observation group and the empty vector group were transfected with silencing cIAP-1 gene plasmid cIAP-1-RNAi and negative gene plasmid NC-RNAi,and the control group was not transfected. The cIAP-1 mRNA and protein levels of cells were detected by real-time PCR and Western blotting. The HS766 T cells with stably silenced cIAP-1 in the observation group and HS766 T cells in the control group were collected to determine the survival rate of cells after 2,4,6 and 8 Gy X ray irradiation for 24 h by CCK-8,as well as the apoptotic rate of cells after6 Gy X ray irradiation for 12,24 and 48 h by flow cytometry. Results The relative expression of cIAP-1 mRNA and protein in the observation group was lower than that in the empty vector group and the control group（P〈0. 05）,and there was no significant difference in the relative expression of cIAP-1 mRNA and protein between the empty vector group and the control group. With the increase of radiation dose,the survival rate of cells decreased; under the same irradiation dose,the cell survival rate in observation group was lower than that in the empty vector group and the control group（P〈0. 05）,and the cell survival rate showed no significant difference between the empty vector and control group. Over the time of irradiation,the apoptotic rate showed an increasing trend; at the same time point,the apoptotic rate of cells in the observation group was higher than that of the empty vector group and the control group（P〈0. 05）,and there was no significant difference between the empty vector and control group. Conclusion Silencing cIAP-1 gene expression can inhibit the survival of pancreatic cancer cells,promote apoptosis and enhance the sensitivity of pancreatic cancer cells to radiotherapy.
出处
《山东医药》
CAS
北大核心
2017年第34期12-15,共4页
Shandong Medical Journal
关键词
胰腺癌
细胞凋亡抑制蛋白1
HS766T细胞
放射疗法
敏感性
pancreatic carcinoma
cellular inhibitor of apoptosis protein 1
HS766T cells
radiotherapy
sensitivity
