本文選題：血清反應因子 + 血清反應因子N端片段��； 參考：《臨床與實驗病理學雜志》2017年10期

【摘要】：目的探討血清反應因子全長(SRF-Full)及N端片段(SRF-N,1-254aa)對TGF-β1誘導c-Kit+心臟干細胞(cardiac stem cell,CSC)分化的影響。方法從c DNA文庫擴增大鼠SRF-Full及SRF-N表達序列并克隆入酶切線性化慢病毒載體GV358(Ubi-MCS-3FLAG-SV40-EGFP-IRES-puromycin),轉化感受態(tài)細胞篩選出陽性克隆并測序鑒定。將SRF-Full及SRF-N過表達質粒與病毒包裝輔助質粒共轉染工具細胞HEK293T,包裝慢病毒。磁激活細胞分選法分離乳SD大鼠c-Kit+CSC,將SRF-Full及SRF-N過表達慢病毒感染CSC并經(jīng)TGF-β1誘導,定量PCR分析下游分化基因mRNA水平的變化。結果成功擴增出SRF-Full及SRF-N編碼序列并正確連接入線性化載體,獲得的陽性轉化子經(jīng)測序無誤。將轉化子質粒與病毒包裝輔助質粒共轉染HEK293T后,獲得滴度為2×108TU/m L的慢病毒顆粒,Western blot檢測到預期Flag融合蛋白。重組慢病毒感染CSC后細胞核中見e GFP信號。TGF-β1誘導CSC出現(xiàn)心肌細胞分化基因(Nkx2.5、Gata4、c Tn I)及平滑肌細胞分化基因(SM22α)的表達,內皮細胞分化(v WF)無影響,SRF-Full促進CSC的心肌細胞分化,而SRF-N則抑制這種分化。結論成功構建SRFFull及SRF-N過表達重組慢病毒質粒。SRF-Full促進而SRF-N減弱TGF-β1誘導CSC的心肌細胞分化。
[Abstract]:Objective to investigate the effects of SRF-Full and N-terminal SRF-NU (1-254 aa) on the differentiation of c-Kit cardiac stem cells induced by TGF- 尾 _ 1. Methods Rat SRF-Full and SRF-N expression sequences were amplified from the cDNA library and cloned into the vector GV358 Ubi-MCS-3FLAG-SV40-EGFP-IRES-puromycinin. The positive clones were screened and sequenced. SRF-Full and SRF-N overexpression plasmids were co-transfected with viral packaging assistant plasmid HEK293T to package lentivirus. The c-Kit CSCs of SD rats were isolated by magnetic activated cell sorting method. SRF-Full and SRF-N overexpression lentiviruses were infected with CSC and induced by TGF- 尾 1. The mRNA levels of downstream differentiation genes were analyzed by quantitative PCR. Results the SRF-Full and SRF-N coding sequences were successfully amplified and correctly ligated into the linearized vector. The obtained positive transformants were sequenced correctly. After the transformant plasmid was co-transfected into HEK293T with virus packaging assistant plasmid, the expected Flag fusion protein was detected by Western blot with a titer of 2 脳 108TU / mL lentivirus particles. E GFP signal. TGF- 尾 1 induced the expression of differentiation gene Nkx2. 5 and smooth muscle cell differentiation gene SM22 偽 in the nucleus of recombinant lentivirus infected with CSC, but endothelial cell differentiation v WF did not affect SRF-Full in promoting the differentiation of CSC cardiomyocytes. SRF-N inhibits this differentiation. Conclusion SRFFull and SRF-N overexpression recombinant lentivirus plasmids. SRF-Full promoted SRF-Full and SRF-N attenuated TGF- 尾 1 induced myocardial differentiation of CSC.
【作者單位】： 廣東醫(yī)科大學基礎醫(yī)學院病理學系;廣東醫(yī)科大學基礎醫(yī)學院病理生理學教研室;海南醫(yī)學院第一附屬醫(yī)院心血管研究所;
【基金】：國家自然科學基金(81460042) 廣東省教育廳科技創(chuàng)新項目基金(KJCX20130088) 廣東省“揚帆計劃”高層次人才項目(4YF16007G) 2015年國家留學人員科技活動擇優(yōu)資助項目
【分類號】：R54

【相似文獻】

相關期刊論文 前10條

1 張玉勤;;細胞分化再活化治療腫瘤[J];國外醫(yī)學情報;1991年01期

2 陳思;黃欣;陳少華;楊力建;鄭海濤;李揚秋;;BCL11B基因在T細胞分化和增殖中調節(jié)作用的研究[J];中國病理生理雜志;2010年10期

3 陳銘珍,葉中綠,蔡康榮,黃秀蘭,陳日玲,程涵蓉;IL-15對MDS患者CD34~+細胞誘導增殖分化和抗凋亡作用[J];中國實驗血液學雜志;2005年04期

4 劉文賓;王兆鉞;;CD4~+ T細胞在慢性ITP中的作用[J];血栓與止血學;2008年03期

5 馬文珠;;心梗不可怕,神奇細胞來幫忙[J];祝您健康;2004年01期

6 汪萍;李曉軍;黃宇烽;賈麗;馬百坤;楊家新;;分化抑制因子基因在血液系統(tǒng)惡性腫瘤細胞中的表達[J];臨床檢驗雜志;2006年02期

7 仲惟昆;;T細胞和B細胞及其臨床意義[J];湖北醫(yī)學院學報;1982年04期

8 周宏;萬海英;李冬;;尿多酸肽誘導急性早幼粒細胞白血病細胞株凋亡的實驗研究[J];同濟大學學報(醫(yī)學版);2012年04期

9 n澘諦閼，

本文編號：2045587