【摘要】：背景與目的:腫瘤放化療的效果與腫瘤細胞凋亡成正相關。Survivin是凋亡抑制基因,在肺癌中過度表達,降低其表達可增加肺癌細胞凋亡。RNA干擾可以特異、高效地封閉該基因的表達。腫瘤組織接受一定劑量的照射后,也可以提高基因轉(zhuǎn)導率。本研究旨在探討預照射聯(lián)合瘤內(nèi)注射si RNA對小鼠肺癌Survivin基因表達的影響。方法:將皮下移植瘤小鼠隨機分為4組:未處理組(A組)、單純si RNA瘤內(nèi)注射組(B組)、單純放射組(C組)和4 Gy預照射+si RNA瘤內(nèi)注射組(D組)。小鼠經(jīng)上述不同處理2 d后用頸椎脫臼法處死,剝離皮下移植瘤,采用實時熒光定量PCR(quantitative real-time PCR,q RT-PCR)和蛋白[質(zhì)]印跡法(Western blot)檢測Survivin基因在m RNA和蛋白水平的表達情況;采用流式細胞術(shù)檢測細胞周期及細胞凋亡。結(jié)果:B組、C組、D組Survivin基因在m RNA和蛋白水平的表達均較A組減少,且D組與A組Survivin基因在m RNA和蛋白水平的差異有統(tǒng)計學意義(P=0.036);D組處于S期的細胞比例為(2.70±0.34)%,較B組[(8.93±0.75)%]和C組[(6.71±0.51)%]明顯減少,且與A組S期細胞的比例相比,差異有統(tǒng)計學意義(P=0.034);D組腫瘤組織的細胞凋亡率為(25.6±0.65)%,較其他3組腫瘤組織的細胞凋亡率明顯增多,差異有統(tǒng)計學意義(F=78.82,P0.05)。結(jié)論:預照射可增強si RNA的轉(zhuǎn)導率,使肺癌組織中Survivin基因表達水平降低,促進細胞凋亡,進而增加放療敏感性。
[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，

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