| 人Notch1受体胞内段重组腺病毒构建及转染 |
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| 引用本文： | 胡彦华,吴德全,姜明山,金光鑫,张雪松,胡彦建,李红影.人Notch1受体胞内段重组腺病毒构建及转染[J].胃肠病学和肝病学杂志,2014(3):319-322. |
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| 作者姓名： | 胡彦华 吴德全 姜明山 金光鑫 张雪松 胡彦建 李红影 |
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| 作者单位： | [1]哈尔滨医科大学附属第二医院普通外科,黑龙江哈尔滨150086 [2]哈尔滨医科大学附属第一医院急诊外科 ,黑龙江哈尔滨150086 [3]哈尔滨医科大学附属第二医院消化内科 ,黑龙江哈尔滨150086 [4]黑龙江中医药大学生化教研室,黑龙江哈尔滨150086 |
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| 基金项目： | 黑龙江省教育厅科学技术研究基金面上项目（12521178） |
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| 摘 要： | 目的构建人Notch1受体胞内段重组腺病毒并转染人肝癌细胞株HepG2,为Notch信号对肝癌侵袭转移的影响研究奠定实验基础。方法构建Notch1受体胞内段(notch intracellular domain,NICD)基因的重组质粒pDC316-CMV-EGFP-CMV-Notch1-Myc,采用AdMax腺病毒包装系统将重组质粒与骨架质粒pBHGlox_E1,3Cre共转染293细胞,同源重组产生重组腺病毒Ad-EGFP-NICD-Myc,RTPCR鉴定重组腺病毒EGFP及NICD基因表达。重组腺病毒转染人肝癌细胞株HepG2,荧光显微镜观察EGFP及NICD蛋白的表达。结果重组腺病毒Ad-EGFP-NICD-Myc经鉴定基因表达正确,此病毒转染人肝癌细胞株HepG2后阳性表达EGFP及NICD蛋白。结论人Notch1受体胞内段重组腺病毒成功构建,为进一步Notch信号对肝癌侵袭转移的影响研究奠定了实验基础。
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| 关 键 词： | Notch信号通路 重组腺病毒 转染 肝癌 治疗 |
Construction and transfection of recombinant adenoviruses with human Notchl re- ceptor intracellular domain gene |
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| HU Yanhua,WU Dequan,JIANG Mingshan,JIN Guangxin,ZHANG Xuesong,HU Yanjian,LI Hongying.Construction and transfection of recombinant adenoviruses with human Notchl re- ceptor intracellular domain gene[J].Chinese Journal of Gastroenterology and Hepatology,2014(3):319-322. |
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| Authors: | HU Yanhua WU Dequan JIANG Mingshan JIN Guangxin ZHANG Xuesong HU Yanjian LI Hongying |
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| Affiliation: | 1. Department of General Surgery, the Second Affiliated Hospital of Harbin Medical University, Harbin 150086 ; 2. De- partment of Emergency Surgery, the First Affiliated Hospital of Harbin Medical University; 3. Department of Gastroen- terology and Hepatology, the Second Affiliated Hospital of Harbin Medical University; 4. Department of Biochemistry, Heilongjiang University of Chinese Medicine, China) |
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| Abstract: | Objective To investigate the effects of Notch signaling on the invasion and metastasis of hepatocellular carcinoma cells through constructing recombinant adenovirus with Notchl receptor intracellular domain and infecting hu- man HepG2 cell lines with the virus. Methods Recombinant shuttle vector pDC316-CMV-EGFP-CMV-NICD-Myc was generated firstly. The Myc-tagged NICD expression recombinant adenoviruses （Ad-EGFP-NICD-Myc） were produced in 293 cells following cotransfection with pDC316-CMV-EGFP-CMV-NICD-Myc shuttle vector and pBHGlox_E1, 3Cre packaging vector. Notch intracellular domain （NICD） gene and enhance green gluorescent protein （EGFP） gene analy- sis of Ad-EGFP-NICD-Myc were taken by RT-PCR. Ad-EGFP-NICD-Myc was transfected into HepG2 in vitro, and then the expression of EGFP and NICD were detected by fluorescence microscope. Results Ad-EGFP-NICD-Myc was suc- cessfully constructed. After infection of Ad-EGFP-NICD-Myc recombinant adenoviruses, the expression of EGFP and NICD in HepG2 were positive. Conclusion Construction of Ad-EGFP-NICD-Myc will provide a basis for a further stud- y of the effect of Notch signaling on the invasion and metastasis of hepatocellular carcinoma ceils. |
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| Keywords: | Notch signaling Recombinant retrovirus Transfection Hepatocellular carcinoma Treatment |
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