本文選題：SWAP-　切入點：神經(jīng)膠質(zhì)瘤　出處：《江蘇醫(yī)藥》2016年21期 　論文類型：期刊論文

【摘要】：目的構(gòu)建下調(diào)及過表達SWAP-70的慢病毒載體,并檢測其在神經(jīng)膠質(zhì)瘤細胞中的表達效果。方法采用PCR擴增目的基因片段,構(gòu)建SWAP-70慢病毒表達載體,與慢病毒轉(zhuǎn)染系統(tǒng)共轉(zhuǎn)染至293T細胞。收集病毒顆粒上清液,感染神經(jīng)膠質(zhì)瘤細胞。熒光顯微鏡下觀察細胞中綠色熒光蛋白的表達情況,并采用Western blot法鑒定SWAP-70蛋白表達水平。結(jié)果成功構(gòu)建下調(diào)及過表達SWAP-70的慢病毒載體,感染神經(jīng)膠質(zhì)瘤細胞的效率達90%左右。Western blot結(jié)果顯示,下調(diào)SWAP-70的神經(jīng)膠質(zhì)瘤細胞中SWAP-70蛋白表達降低(P0.01),而過表達SWAP-70的神經(jīng)膠質(zhì)瘤細胞中內(nèi)源性及外源性SWAP-70蛋白均有表達。結(jié)論成功建立下調(diào)及過表達SWAP-70的慢病毒載體,并有效轉(zhuǎn)染神經(jīng)膠質(zhì)瘤細胞,為膠質(zhì)瘤的基因治療研究提供參考。
[Abstract]:Objective to construct a lentivirus vector for down-regulation and overexpression of SWAP-70 and to detect its expression in glioma cells. Methods SWAP-70 lentivirus expression vector was constructed by PCR amplification of the target gene fragment. Co-transfection with lentivirus transfection system to 293T cells. The supernatant of virus particles was collected to infect glioma cells. The expression of green fluorescent protein (GFP) in the cells was observed under fluorescence microscope. The expression level of SWAP-70 protein was identified by Western blot. Results Lentivirus vector was successfully constructed for down-regulation and overexpression of SWAP-70. The efficiency of infected glioma cells was about 90%. The down-regulation of SWAP-70 protein expression in glioma cells with SWAP-70 decreased the expression of SWAP-70 protein, while the expression of endogenous and exogenous SWAP-70 protein was found in glioma cells with overexpression of SWAP-70. Conclusion Lentivirus vector for down-regulation and overexpression of SWAP-70 is successfully established. And transfection of glioma cells to provide a reference for gene therapy of glioma.
【作者單位】： 徐州醫(yī)科大學(xué)研究生學(xué)院;徐州醫(yī)科大學(xué)神經(jīng)系統(tǒng)疾病研究所;徐州醫(yī)科大學(xué)附屬醫(yī)院神經(jīng)外科;
【基金】：國家自然科學(xué)基金青年基金(81402074)
【分類號】：R739.41

【相似文獻】

相關(guān)期刊論文 前10條

1 陳贊,楊立莊,葉偉,王伊龍,趙鵬,李永利,康軍,鄭永日,曹陽;神經(jīng)膠質(zhì)瘤細胞熱休克蛋白抗原肽復(fù)合物的研究[J];中華實驗外科雜志;2003年05期

2 張玲華;;神經(jīng)膠質(zhì)瘤的診斷與治療現(xiàn)狀[J];實用醫(yī)技雜志;2007年24期

3 ;顱面神經(jīng)膠質(zhì)瘤[J];國外醫(yī)學(xué).耳鼻咽喉科學(xué)分冊;1997年02期

4 薛德麟,雷霆;要重視神經(jīng)膠質(zhì)瘤的治療[J];中國臨床神經(jīng)外科雜志;2002年01期

5 肖敏 ,吳淼 ,曾冬竹 ,饒蕓;樹突狀細胞瘤苗誘導(dǎo)的抗神經(jīng)膠質(zhì)瘤作用體外實驗研究[J];現(xiàn)代醫(yī)藥衛(wèi)生;2003年01期

6 戴捷,于\，

本文編號：1606780