本文關(guān)鍵詞： RNA干擾 MCF-7細胞 小干擾RNA 血管內(nèi)皮生長因子 survivin　出處：《青島大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：探討靶向血管內(nèi)皮生長因子(VEGF)的siRNA下調(diào)VEGF表達后對人乳腺癌MCF-7細胞凋亡及凋亡相關(guān)基因survivin的影響 方法：體外設(shè)計并合成靶向VEGF mRNA的siRNA,采用LipofectamineTM2000轉(zhuǎn)染人乳腺癌細胞MCF-7,實驗設(shè)空白對照組、空脂質(zhì)體組、陰性對照SCR組、3個siRNA濃度梯度組(50、100、200nnmol/L),轉(zhuǎn)染24小時后,AnnexinV/PI雙染結(jié)合流式細胞儀測定細胞凋亡；半定量RT-PCR、細胞免疫組化技術(shù)檢測轉(zhuǎn)染前后MCF-7細胞survivin基因表達的變化。 結(jié)果：靶向VEGF基因的siRNA轉(zhuǎn)染MCF-7后,與空白對照組比較,50nmol/L組細胞凋亡率增加(p0.05),100、200nmol/L組細胞凋亡率顯著增加(p0.01),但空脂質(zhì)體組、陰性對照SCR組與空白對照組比較無統(tǒng)計學(xué)意義(p0.05);survivin mRNA在各siRNA轉(zhuǎn)染組表達較之空白對照組均下調(diào)(p均0.05,其中100nmol/L和200nmol/L轉(zhuǎn)染組p均0.01),空脂質(zhì)體組、陰性對照SCR組與空白對照組比較無統(tǒng)計學(xué)意義(p0.05)；survivin蛋白表達水平在各濃度轉(zhuǎn)染組下調(diào)(p均0.05,其中100nmol/L和200nmol/L轉(zhuǎn)染組p均0.01),而與未處理組比較,陰性對照SCR組與空脂質(zhì)體組差異無統(tǒng)計學(xué)意義(p0.05)。 結(jié)論：靶向VEGF基因的siRNA轉(zhuǎn)染MCF-7后誘導(dǎo)細胞凋亡,且與轉(zhuǎn)染劑量有一定依賴關(guān)系；其分子機制與凋亡相關(guān)基因survivin表達有關(guān)。
[Abstract]:Objective: to investigate the effect of siRNA targeting vascular endothelial growth factor (VEGF) on apoptosis and apoptosis-related gene survivin in human breast cancer MCF-7 cells after down-regulation of VEGF expression. Methods: siRNAs targeting VEGF mRNA were designed and synthesized in vitro. Human breast cancer cell line MCF-7 was transfected with LipofectamineTM2000. In the negative control SCR group, three siRNA concentration gradient groups were divided into three groups. After 24 hours of transfection, Annexin V / Pi double staining combined with flow cytometry was used to detect apoptosis, and semi-quantitative RT-PCR was used to detect the changes of survivin gene expression in MCF-7 cells before and after transfection. Results: after transfection of siRNA targeting VEGF gene into MCF-7, the apoptotic rate of 50 nmol / L group increased significantly compared with that of blank control group, but the apoptosis rate increased significantly in the 200nmol / L group, but in the empty liposome group. There was no significant difference between the negative control group and the blank control group in the expression of survivin mRNA in each siRNA transfection group. The expression of survivin mRNA in each siRNA transfection group was significantly lower than that in the blank control group (P < 0.05), including 100nmol / L and 200nmol / L transfection group (p < 0.01) and empty liposome group (P < 0.05). There was no significant difference between the negative control group and the blank control group in the expression of p0.05 SCR protein. The down-regulated expression level of the protein in each concentration of transfection group was 0.05, including 100nmol / L and 200nmol / L transfection group (p 0.01), but compared with the untreated group (P < 0.05). There was no significant difference between SCR group and empty liposome group in negative control group (p 0.05). Conclusion: siRNA targeting VEGF gene can induce apoptosis of MCF-7 cells in a dose-dependent manner, and its molecular mechanism is related to the expression of apoptosis-related gene survivin.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R346

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