本文關鍵詞： 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》2011年碩士論文　論文類型：學位論文

【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基因survivin的影響 方法：體外設計并合成靶向VEGF mRNA的siRNA,采用LipofectamineTM2000轉染人乳腺癌細胞MCF-7,實驗設空白對照組、空脂質體組、陰性對照SCR組、3個siRNA濃度梯度組(50、100、200nnmol/L),轉染24小時后,AnnexinV/PI雙染結合流式細胞儀測定細胞凋亡；半定量RT-PCR、細胞免疫組化技術檢測轉染前后MCF-7細胞survivin基因表達的變化。 結果：靶向VEGF基因的siRNA轉染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),但空脂質體組、陰性對照SCR組與空白對照組比較無統(tǒng)計學意義(p0.05);survivin mRNA在各siRNA轉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轉染組p均0.01),空脂質體組、陰性對照SCR組與空白對照組比較無統(tǒng)計學意義(p0.05)；survivin蛋白表達水平在各濃度轉染組下調(diào)(p均0.05,其中100nmol/L和200nmol/L轉染組p均0.01),而與未處理組比較,陰性對照SCR組與空脂質體組差異無統(tǒng)計學意義(p0.05)。 結論：靶向VEGF基因的siRNA轉染MCF-7后誘導細胞凋亡,且與轉染劑量有一定依賴關系；其分子機制與凋亡相關基因survivin表達有關。
[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.
【學位授予單位】：青島大學
【學位級別】：碩士
【學位授予年份】：2011
【分類號】：R346

【參考文獻】

相關期刊論文 前6條

1 孫志清;項鋒鋼;王文華;;人腦星形細胞瘤組織Survivin與VEGF表達及意義[J];齊魯醫(yī)學雜志;2008年06期

2 閻錫蘊;;腫瘤新生血管及分子靶向治療新策略[J];生物物理學報;2010年03期

3 張曉;徐文華;葛銀林;侯琳;李泉;;VEGF基因特異性siRNA轉染后對人乳腺癌細胞增殖和凋亡的影響[J];細胞與分子免疫學雜志;2007年01期

4 張曉靜;葛銀林;侯琳;李泉;;KDR為靶的siRNA抑制乳腺癌細胞增殖的體內(nèi)外研究[J];細胞與分子免疫學雜志;2008年01期

5 賈如江;侯麗艷;張有成;呂志誠;;VEGFR-3 siRNA體外對人結腸癌細胞生長的抑制作用[J];世界華人消化雜志;2008年16期

6 賈如江;侯麗艷;劉奇;馮運章;;VEGFR-3 siRNA對人結腸癌細胞黏附力和侵襲性的抑制作用[J];中國腫瘤生物治療雜志;2008年06期

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