本文關(guān)鍵詞： c-Myc 反轉(zhuǎn)錄病毒載體 過表達 增殖 凋亡　出處：《東北林業(yè)大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：c-Myc為原癌基因,為正常組織器官發(fā)育過程中所必需。它主要通過特異或非特異性與靶基因DNA結(jié)合位點相結(jié)合,啟動或抑制靶基因的表達,調(diào)節(jié)細胞生長、分化等生命活動。有研究發(fā)現(xiàn),c-Myc基因在染色體上發(fā)生擴增或激活突變會引起細胞增殖加快,細胞侵襲能力加強,導(dǎo)致胰腺腫瘤的發(fā)生、發(fā)展及惡化；在p細胞中過表達c-Myc也可誘導(dǎo)細胞凋亡,減少β細胞群體數(shù)量。為了進一步研究c-Myc對于p細胞的調(diào)控機制,本研究通過用293GP細胞包裝帶有c-Myc基因的逆轉(zhuǎn)錄病毒載體,制備假性逆轉(zhuǎn)錄病毒,用其侵染胰腺β細胞系MIN6。通過對整合c-Myc基因的MIN6細胞G418篩選、擴增獲得穩(wěn)定過表達c-Myc基因的MIN6細胞克隆,并進一步分析c-Myc過表達引起MIN6細胞的生命活動變化。 結(jié)果表明,通過抗性藥物G418的篩選,獲得整合逆轉(zhuǎn)錄病毒基因的多個MIN6細胞克隆RT-PCR檢測結(jié)果表明,這些細胞能穩(wěn)定過表達c-Myc基因,同時發(fā)現(xiàn),在過表達c-Myc的M1N6細胞中可以檢測到mtR-17-92基因簇的表達上調(diào)；通過分析過表達c-Myc克隆細胞的生長曲線,我們發(fā)現(xiàn),過表達c-Myc會引起MIN6細胞生長加快,細胞周期變短；通過細胞染色,我們發(fā)現(xiàn)某些過表達c-Myc的MIN6細胞凋亡有明顯增加。這些研究結(jié)果表明,在小鼠β細胞系中穩(wěn)定過表達c-Myc會誘導(dǎo)MIN6細胞增殖加快、凋亡加劇,其機制有待于進一步的實驗探索。
[Abstract]:C-Myc is a proto-oncogene that is necessary for the development of normal tissues and organs. It activates or inhibits the expression of target genes and regulates cell growth through the binding of specific or non-specific DNA binding sites to target genes. Some studies have found that the amplification or activation of c-Myc gene on chromosomes will accelerate cell proliferation, enhance the ability of cell invasion, and lead to the occurrence, development and deterioration of pancreatic tumors. Overexpression of c-Myc in p cells also induced apoptosis and reduced the number of 尾 cells. In order to further study the regulation mechanism of c-Myc on p cells, the retrovirus vector with c-Myc gene was packaged with 293GP cells. Pseudoretrovirus was prepared and infected with human pancreatic 尾 cell line MIN6.The MIN6 cell clones with stable overexpression of c-Myc gene were amplified by G418 screening of MIN6 cells integrated with c-Myc gene. Furthermore, the changes of life activity of MIN6 cells induced by c-Myc overexpression were further analyzed. The results showed that through the screening of the resistant drug G418, RT-PCR clones of multiple MIN6 cells integrating retrovirus gene were obtained. The results showed that these cells were able to overexpression c-Myc gene stably, and found that, Overexpression of c-Myc in M1N6 cells can be detected by up-regulation of mtR-17-92 gene cluster. By analyzing the growth curve of overexpression c-Myc clone cells, we found that overexpression of c-Myc can accelerate the growth of MIN6 cells and shorten cell cycle. We found that the apoptosis of some MIN6 cells with overexpression of c-Myc was significantly increased. These results indicated that overexpression of c-Myc in mouse 尾 cell lines could induce the proliferation and apoptosis of MIN6 cells, and the mechanism needed to be further explored.
【學(xué)位授予單位】：東北林業(yè)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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