杨小燕,金皎,黄璟,许键炜,吴莎莎,马健娟,李燕,庹媛媛,杨红兰,潘海新,胡绍燕,何志旭
YANG Xiaoyan,JIN Jiao,HUANG Jing,XU Jianwei,WU Shasha,MA Jianjuan,LI Yan,TUO Yuanyuan,YANG Honglan,PAN Haixin,HU Shaoyan,HE Zhixu

• 1:贵州医科大学附院
• 2:贵州医科大学干细胞与组织工程实验中心
• 3:苏州大学附属儿童医院

摘要(Abstract)：

目的:构建人S100A8基因高表达的重组慢病毒表达载体。方法:以正常人骨髓单个核细胞的c DNA为扩增模板,克隆S100A8基因全长的c DNA,与p MD19-T载体连接后进行测序鉴定;将p LVX-IRES-puro目的载体与测序正确的S100A8片段进行连接,筛选阳性克隆后进行菌落PCR、双酶切及测序鉴定。结果:p LVXS100A8重组质粒双酶切及菌落PCR产物与目的基因相关片段S100A8 c DNA大小一致,成功克隆S100A8基因,S100A8基因成功插入到p LVX-IRES-puro。结论:成功构建了人S100A8基因高表达的重组慢病毒表达载体。
Objective: To construct the recombinant lentiviral expression vector of human S100 A8 gene high expression. Methods: The marrow mononuclear cells c DNA as template in the normal human bone,the full-length c DNA of cloned S100 A8 gene was connected to the p MD19-T vector and identified by sequencing. The p LVX-IRES-puro target vector was connected with the sequenced S100 A8 fragment. After screening the positive clones,colony PCR,double enzyme digestion and sequencing were performed. Results: PLVX-S100 A8 recombinant plasmid double enzyme digestion and colony PCR products have the same size of target gene fragments related to S100 A8 c DNA. The S100 A8 gene was successfully cloned and inserted into p LVX-IRES-puro. Conclusions: The recombinant lentiviral expression vector of human S100 A8 gene high expression is successfully constructed.

关键词(KeyWords)： 基因;克隆细胞;高表达;慢病毒载体;构建
gene;clone cell;overexpression;lentiviral vector;construction

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅联合基金[黔科合LH(2016)7240号]

作者(Author): 杨小燕,金皎,黄璟,许键炜,吴莎莎,马健娟,李燕,庹媛媛,杨红兰,潘海新,胡绍燕,何志旭
YANG Xiaoyan,JIN Jiao,HUANG Jing,XU Jianwei,WU Shasha,MA Jianjuan,LI Yan,TUO Yuanyuan,YANG Honglan,PAN Haixin,HU Shaoyan,HE Zhixu

DOI: 10.19367/j.cnki.1000-2707.2017.11.007

参考文献(References)：

• [1]Kerkhoff C,Klempt M,Sorg C.Novel insights into structure and function of MRP8(S100A8)and MRPl4(S100A9)[J].Biochim Biophy Acta,1998(2):200-211.
• [2]Gebhardt C,Németh J,Angel P,et al.S100A8 and S100A9 in inflammation and cancer[J].Biochem Pharmacol,2006(11):1622-1631.
• [3]Geetha S.S100A8 and S100A9:New Insights into their roles in malignancy[J].Journal of Innate Immunity,2012(4):31-40.
• [4]张学梅,艾飞艳,沈守荣.外源性S100a8和S100a9与结肠癌侵袭转移的关系研究[J].医学信息,2014(4):312-313.
• [5]Rhee DK,Park SH,Jang KY.Molecular signatures associated with transformation and progression to breast cancer in the isogenic MCF10 model[J].Genomics,2008(6):419-428.
• [6]Grebhardt S,Müller-Decker K,Bestvater F,et al.Impact of S100A8/S100A9expression on prostate cancer progression in vitro and in vivo[J].J Cell Physiol,2014(5):661-671.
• [7]Qin F,Song Y,Li Z,et al.S100A8/A9 induces apoptosis and inhibits metastasis of Cas Ki human cervical cancer cells[J].Pathol Oncol Res,2010(3):353-360.
• [8]Wan SG,Taccioli C,Jiang Y,et al.Zinc deficiency activates S100A8 inflammation in the absence of COX-2 and promotes murine oral-esophagealtumor progression[J].Int J Cancer,2011(2):331-345.
• [9]De Ponti A,Wiechert L,Schneller D,et al.A pro-tumorigenic function of S100A8/A9 in carcinogen-induced hepatocellular carcinoma[J].Cancer Lett,2015(2):396-404.344-351.
• [10]Spijkers-Hagelstein JA,Schneider P,Hulleman E,et al.Elevated S100A8/S100A9 expression causes glucocorticoid resistance in MLL-rearranged infant acute lymphoblastic leukemia[J].Leukemia,2012(6):1255-1265.
• [11]Yang L,Yang M,Zhang H,et al.S100A8-targeting siRNA enhances arsenic trioxide-induced myeloid leukemia cell death by down-regulating autophagy[J].Int J Mol Med,2012(1):65-72.
• [12]Yang M,Zeng P,Kang R,et al.S100A8 contributes to drug resistance by promoting autophagy in leukemia cells[J].PLo S One,2014(5):e97242.

文章评论(Comment)：