人DLL1真核表达质粒的构建及过表达DLL1抑制人口腔鳞癌细胞SCC15的增殖

发布时间：2018-04-10 02:26

本文选题：Notch信号通路　切入点：人DLL　出处：《暨南大学学报(自然科学与医学版)》2017年03期

【摘要】：目的:构建人全长DLL1基因真核表达质粒及过表达DLL1对人口腔鳞癌细胞增殖的影响.方法:PCR扩增人全长DLL1基因,酶切和测序鉴定后克隆至真核表达载体pCMV-Tag4并构建真核重组质粒DLL1/pC MVTag4,瞬时转染HEK293T细胞并通过实时荧光定量PCR(Q-PCR)和Western blot鉴定DLL1的表达;进一步检测DLL1在SCC15等3株人口腔鳞癌细胞中的表达;DLL1/pCMV-Tag4瞬时转染SCC15细胞,Q-PCR和Western blot检测过表达及MTT检测DLL1过表达对细胞SCC15增殖的影响.结果:真核重组质粒DLL1/pCMV-Tag4在HEK293T细胞中成功表达;DLL1在口腔鳞癌细胞中的表达高于正常口腔细胞;DLL1/pCMV-Tag4在SCC15细胞中获得表达且过表达DLL1抑制SCC15细胞的增殖.结论:人全长DLL1分子在HEK293T及SCC15细胞中获得表达,过表达DLL1抑制口腔鳞癌细胞SCC15的增殖.
[Abstract]:Aim: to construct the eukaryotic expression plasmid of human full-length DLL1 gene and the effect of overexpression of DLL1 on the proliferation of human oral squamous cell carcinoma cells.Methods the full-length human DLL1 gene was amplified by 1: PCR and cloned into eukaryotic expression vector pCMV-Tag4 after restriction endonuclease digestion and sequencing. The eukaryotic recombinant plasmid DLL1/pC MV Tag4 was constructed. The recombinant plasmid DLL1/pC MV Tag4 was transiently transfected into HEK293T cells. The expression of DLL1 was identified by real-time fluorescence quantitative PCR PCR Q-PCR and Western blot.The expression of DLL1 in three human oral squamous cell carcinoma (SCC15) cells was further detected by Q-PCR, Western blot and DLL1 overexpression in SCC15 cells transfected with DLL1 / pCMV-Tag4 and the effect of MTT on SCC15 proliferation.Results: the expression of DLL1 in HEK293T cells was higher than that in normal oral squamous cell carcinoma cells (DL1 / pCMV-Tag4), and the overexpression of DLL1 inhibited the proliferation of SCC15 cells.Conclusion: human full-length DLL1 was expressed in HEK293T and SCC15 cells, and overexpression of DLL1 inhibited the proliferation of SCC15 in oral squamous carcinoma cells.
【作者单位】：南方医科大学检验与生物技术学院生物治疗研究所;珠海南医大生物医药公共服务平台有限公司;
【基金】：国家高技术研究发展技术(863计划)项目(2014AA020909) 国家自然科学基金-广东省联合基金重点项目(U1401223) 国家自然科学基金面上项目(81470831) 广东省科技计划创新载体建设项目(2013B090800036);广东省科技计划项目(2015A040404021;2016B090919019)
【分类号】：R739.8

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