靶向性RNA干擾對(duì)MCF-7細(xì)胞與MSC細(xì)胞hTERT基因表達(dá)和細(xì)胞生長的影響.pdf

1、瀘州醫(yī)學(xué)院碩士學(xué)位論文靶向性RNA干擾對(duì)MCF7細(xì)胞與MSC細(xì)胞hTERT基因表達(dá)和細(xì)胞生長的影響姓名：段軼鋆申請學(xué)位級(jí)別：碩士專業(yè)：腫瘤分子遺傳學(xué)指導(dǎo)教師：稅青林20090501為攜帶DF3啟動(dòng)子而無hTERT干擾序列的質(zhì)粒pGenesil10mir30DF3shRNAHK：CMV對(duì)照組為攜帶CMV啟動(dòng)子的hTERT干擾質(zhì)粒pGenesil10mir30CMVshRNAhTERT；空白對(duì)照組僅含等量PBS液。以構(gòu)建的質(zhì)粒分別轉(zhuǎn)染乳腺癌

2、MCF7細(xì)胞和間充質(zhì)干細(xì)胞。轉(zhuǎn)染后12h，24h，48h，72h通過流式細(xì)胞儀檢測細(xì)胞生長凋亡情況；MTT法檢測細(xì)胞增殖情況；于轉(zhuǎn)染后48hELISA法檢測hTERT蛋白表達(dá)情況。結(jié)果：1重組質(zhì)粒pGenesil10mir30DF3shRNAhTERT經(jīng)酶切鑒定分析證實(shí)目的序列己插入到預(yù)計(jì)位點(diǎn)，符合設(shè)計(jì)要求，重組質(zhì)粒構(gòu)建成功。2重組質(zhì)粒pGenesil10mir30DF3shRNAhTERT酶切鑒定分析證實(shí)DF3啟動(dòng)子已經(jīng)成功替代原質(zhì)粒

3、中CMV啟動(dòng)子。3ELISA法檢測結(jié)果顯示：乳腺癌MCF7細(xì)胞的實(shí)驗(yàn)組和CMV對(duì)照組hTERT蛋白濃度顯著下降。同時(shí)MSC細(xì)胞CMV對(duì)照組hTERT蛋白濃度顯著下降而實(shí)驗(yàn)組hTERT蛋白濃度無顯著變化。4MTT結(jié)果顯示：乳腺癌MCF7細(xì)胞的實(shí)驗(yàn)組和CMV對(duì)照組細(xì)胞在2472h范圍內(nèi)細(xì)胞增殖顯著抑制；而MSC細(xì)胞僅在CMV對(duì)照組增殖顯著抑制，MSC細(xì)胞實(shí)驗(yàn)組細(xì)胞生長無顯著抑制。5流式細(xì)胞儀檢測細(xì)胞凋亡乳腺癌MCF7細(xì)胞實(shí)驗(yàn)組和CMV對(duì)照組