全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

携SH2-Caspase8融合基因重组腺病毒的构建及对K562细胞增殖的影响

, PP. 1887-1892

Keywords: 细胞增殖,SH,Caspase,重组腺病毒,构建,K细胞,bcr-abl?

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的构建并鉴定携SH2-Caspase8融合基因的重组腺病毒AdE-SC-EGFP及其突变体,观察其对K562细胞增殖的抑制作用。方法采用RT-PCR、重叠PCR扩增SH2-Caspase8融合基因,克隆至腺病毒穿梭载体pAdTrack-CMV中,构建穿梭质粒pAdT-SC-EGFP,进行酶切和测序鉴定。将PmeⅠ酶切后的穿梭质粒转化pAdEasy-BJ5183感受态,通过细菌内同源重组产生复制缺陷型重组腺病毒质粒pAdE-SC-EGFP及其突变体,?PacⅠ酶切后将其转染AD293细胞进行包装,PCR和Westernblot鉴定重组腺病毒AdE-SC-EGFP,验证后扩增病毒并测定滴度。结果PCR检测、酶切和测序结果均表明腺病毒穿梭质粒pAdT-SC-EGFP和重组腺病毒质粒pAdE-SC-EGFP构建成功;PCR检测、EGFP表达检测结果证明重组腺病毒AdE-SC-EGFP包装成功,扩增后,滴度可达1.5×1012pfu/ml。感染K562细胞后,Westernblot检测目的蛋白的表达,MTT和半固体集落形成实验检测其对K562细胞生长增殖的影响。结论成功构建并鉴定了携带SH2-Caspase8融合基因的重组腺病毒AdE-SC-EGFP及其突变体;MTT和克隆形成实验证明重组腺病毒AdE-SC-EGFP对BCR-ABL阳性的K562细胞有明显的增殖抑制作用。

References

[1]  刘双凤, 胡晶, 曹唯希, 等. 携SH2-Caspase8融合基因重组腺病毒的构建及对K562细胞增殖的影响[J].第三军医大学学报,2011,33(18):1887-1892.
[2]  周坤,杨辉,周伟,等.药物阻断mGlu3受体通路对脑胶质瘤干细胞增殖与分化的影响[J].第三军医大学学报,2012,34(17):1701.  Zhou Kun,Yang Hui,Zhou Wei,et al.Effect of drug blockade of type-3 metabotropic glutamate receptor on proliferation and differentiation in glioma stem cells[J].J Third Mil Med Univ,2012,34(18):1701. [2]林力,邓碧,寿铸,等.硫链丝菌肽对人喉癌Hep-2细胞生长及FoxM1表达的影响[J].第三军医大学学报,2012,34(20):2086.  Lin Li,Deng Bi,Shou Zhu,et al.Effect of thiostrepton on proliferation and FoxM1 expression in human laryngocarcinoma Hep-2 cell line[J].J Third Mil Med Univ,2012,34(18):2086. [3]王恒,王勇,江晓霁,等.莪术醇通过JAK3/STAT信号途径抑制Jurkat细胞增殖[J].第三军医大学学报,2013,35(05):408.  Wang Heng,Wang Yong,Jiang Xiaoji,et al.Proliferation-inhibiting effect of curcumol on human T-cell leukemia cell line Jurkat via blocking JAK3/STAT pathway[J].J Third Mil Med Univ,2013,35(18):408. [4]邵新宏,于游,张才全.miR-34a靶向调控NOTCH1基因对SW480细胞增殖的影响[J].第三军医大学学报,2012,34(22):2297.  Shao Xinhong,Yu You,Zhang Caiquan.Inhibitory effects of miR-34a on NOTCH1 gene expression and SW480 cell proliferation[J].J Third Mil Med Univ,2012,34(18):2297. [5]倪振洪,王滨,丁雯,等.左旋棉酚通过ERK通路诱导Daudi细胞发生自噬及意义[J].第三军医大学学报,2012,34(23):2384.  Ni Zhenhong,Wang Bin,Ding Wen,et al.(-)-gossypol induces autophagy in Daudi cells through ERK signal pathway[J].J Third Mil Med Univ,2012,34(18):2384. [6]赵亮,李静,魏强,等.人PGI基因siRNA慢病毒质粒的构建及对白血病细胞增殖的影响[J].第三军医大学学报,2013,35(01):20.  Zhao Liang,Li Jing,Wei Qiang,et al.Construction of a lentiviral vector expressing human phosphoglucose isomerase gene siRNA and its influence on leukemia cell proliferation[J].J Third Mil Med Univ,2013,35(18):20. [7]郑英如,李力,刘宝英,等.人可溶性增殖诱导配体促HeLa细胞增殖作用的研究[J].第三军医大学学报,2007,29(20):1951.  ZHENG Ying-ru,LI Li,LIU Bao-ying,et al.Soluble a proliferation-induced ligand promotes proliferation of HeLa cells[J].J Third Mil Med Univ,2007,29(18):1951. [8]夏珊,李世荣,刘剑毅,等.大黄素对人增生性瘢痕成纤维细胞的增殖抑制及细胞内游离Ca2+的作用[J].第三军医大学学报,2007,29(20):1982.  XIA Shan,LI Shi-rong,LIU Jian-yi,et al.Effect of emodin on proliferation and intracellular free calcium of hypertrophic scar fibroblasts in vitro[J].J Third Mil Med Univ,2007,29(18):1982. [9]左国伟,何焕玲,李星星,等.腺病毒介导BMP-2和BMP-6对人骨肉瘤细胞株MG63和U2OS的作用研究[J].第三军医大学学报,2007,29(18):1768.  ZUO Guo-wei,HE Huan-ling,LI Xing-xing,et al.Effects of recombinant BMP-2 and BMP-6 adenoviruses on human osteosarcoma cell line MG63 and U2OS[J].J Third Mil Med Univ,2007,29(18):1768. [10]冉擘力,何国祥,王耿,等.p27基因对血管平滑肌细胞中连接蛋白43表达的影响[J].第三军医大学学报,2008,30(05):424.  RAN Bo-li,HE Guo-xiang,WANG Geng,et al.Effect of p27 gene on expression of connexin43 in vascular smooth muscle cells[J].J Third Mil Med Univ,2008,30(18):424.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133