预照射联合siRNA抑制小鼠肺癌Survivin基因表达的研究
发布时间：2019-06-01 11:01
【摘要】：背景与目的:肿瘤放化疗的效果与肿瘤细胞凋亡成正相关。Survivin是凋亡抑制基因,在肺癌中过度表达,降低其表达可增加肺癌细胞凋亡。RNA干扰可以特异、高效地封闭该基因的表达。肿瘤组织接受一定剂量的照射后,也可以提高基因转导率。本研究旨在探讨预照射联合瘤内注射si RNA对小鼠肺癌Survivin基因表达的影响。方法:将皮下移植瘤小鼠随机分为4组:未处理组(A组)、单纯si RNA瘤内注射组(B组)、单纯放射组(C组)和4 Gy预照射+si RNA瘤内注射组(D组)。小鼠经上述不同处理2 d后用颈椎脱臼法处死,剥离皮下移植瘤,采用实时荧光定量PCR(quantitative real-time PCR,q RT-PCR)和蛋白[质]印迹法(Western blot)检测Survivin基因在m RNA和蛋白水平的表达情况;采用流式细胞术检测细胞周期及细胞凋亡。结果:B组、C组、D组Survivin基因在m RNA和蛋白水平的表达均较A组减少,且D组与A组Survivin基因在m RNA和蛋白水平的差异有统计学意义(P=0.036);D组处于S期的细胞比例为(2.70±0.34)%,较B组[(8.93±0.75)%]和C组[(6.71±0.51)%]明显减少,且与A组S期细胞的比例相比,差异有统计学意义(P=0.034);D组肿瘤组织的细胞凋亡率为(25.6±0.65)%,较其他3组肿瘤组织的细胞凋亡率明显增多,差异有统计学意义(F=78.82,P0.05)。结论:预照射可增强si RNA的转导率,使肺癌组织中Survivin基因表达水平降低,促进细胞凋亡,进而增加放疗敏感性。
[Abstract]:Background & objective: the effect of radiotherapy and chemotherapy is positively correlated with apoptosis of tumor cells. Survivin is an apoptosis inhibitor gene, which is overexpressed in lung cancer and its expression can increase apoptosis of lung cancer cells. RNA interference can be specific. The expression of the gene was blocked efficiently. After a certain dose of irradiation, the gene transfer rate of tumor tissue can also be improved. The purpose of this study was to investigate the effect of pre-irradiation combined with intratumoral injection of si RNA on Survivin gene expression in mouse lung cancer. Methods: subcutaneous tumor transplantation mice were randomly divided into 4 groups: untreated group (group A), simple si RNA intratumoral injection group (group B), simple radiation group (group C) and 4 Gy pre-irradiation si RNA intratumoral injection group (group D). After 2 days of treatment, the mice were killed by cervical dislocated method. The tumor was peeled off and real-time fluorescence quantitative PCR (quantitative real-time PCR, was used. Q RT-PCR) and protein (Western blot) were used to detect the expression of Survivin gene at m RNA and protein level. Cell cycle and apoptosis were detected by flow cytometry. Results: the expression of Survivin gene at m RNA and protein level in group B, group C and group D was lower than that in group A, and the difference in m RNA and protein level between group D and group A was statistically significant (P 鈮，
本文编号：2490221
[Abstract]:Background & objective: the effect of radiotherapy and chemotherapy is positively correlated with apoptosis of tumor cells. Survivin is an apoptosis inhibitor gene, which is overexpressed in lung cancer and its expression can increase apoptosis of lung cancer cells. RNA interference can be specific. The expression of the gene was blocked efficiently. After a certain dose of irradiation, the gene transfer rate of tumor tissue can also be improved. The purpose of this study was to investigate the effect of pre-irradiation combined with intratumoral injection of si RNA on Survivin gene expression in mouse lung cancer. Methods: subcutaneous tumor transplantation mice were randomly divided into 4 groups: untreated group (group A), simple si RNA intratumoral injection group (group B), simple radiation group (group C) and 4 Gy pre-irradiation si RNA intratumoral injection group (group D). After 2 days of treatment, the mice were killed by cervical dislocated method. The tumor was peeled off and real-time fluorescence quantitative PCR (quantitative real-time PCR, was used. Q RT-PCR) and protein (Western blot) were used to detect the expression of Survivin gene at m RNA and protein level. Cell cycle and apoptosis were detected by flow cytometry. Results: the expression of Survivin gene at m RNA and protein level in group B, group C and group D was lower than that in group A, and the difference in m RNA and protein level between group D and group A was statistically significant (P 鈮，
本文编号：2490221
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