人肠凝集素-1真核表达载体的构建及其在胃癌细胞中的表达  

Construction of eukaryotic expression vector of human intelectin-1 and its expression in gastric cancer cells

作　　者：齐盟[1] 张桓瑜[1] 郑丽端[2] 戚腾[1] 童强松[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院外科,武汉430022 [2]华中科技大学同济医学院附属协和医院病理科,武汉430022

出　　处：《中华实验外科杂志》2012年第3期388-390,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30200284、30600278、30772359、81071997、81072073）;教育部新世纪优秀人才支持计划资助项目（NCET-06-0641）;教育部回国人员基金资助项目（2008889）

摘　　要：目的构建人肠凝集素-1（ITLN-1）基因的真核表达载体，观察其在胃癌细胞中的表达。方法采用逆转录聚合酶链反应（1iT-PCR）法扩增ITLN-1eDNA，定向克隆至真核表达载体pEGFP—N1，测序鉴定；在脂质体的介导下，重组质粒瞬时转染人胃癌细胞株SGC-790172h后，West—ernblot法检测细胞培养上清中ITLN．1表达，四甲基偶氮唑盐比色（MTr）法、Transwell小室实验检测癌细胞增殖、侵袭活性。结果真核表达载体pEGFP—ITLN-1转染SGC-7901细胞72h后，ITLN-1蛋白表达上调5．72倍（P〈0．01），细胞增殖活性降低52．8％（P〈0．01），细胞侵袭能力下调58．1％（P〈0．01）。结论成功构建人ITLN-1真核表达载体，转染胃癌细胞过表达后，能抑制癌细胞的增殖和侵袭活性。Objective To construct the eukaryotic expression vector of human intetectin-1 （1TLN-1） and observe its expression in gastric cancer cells. Methods ITLN-1 cDNA was amplified by reverse tran- scription polymerase chain reaction （RT-PCR） , inserted into eukaryotic vector pEGFP-N1, and validated by nucleic acid sequencing. Under the induction of Lipofectamine 2000, the recombinant was transfected into SGC-7901 cells for 72 hours. The expression level of ITLN-1 in medium supernatant was detected by western blotting. 2-（4, 5-dimethyhriazol-2-yl）-2, 5-diphenyl tetrazolium bromide （MTI＇） colorimetry and transwell assay were applied to measure the in vitro proliferation and invasion of cancer cells. Results Seventy-two hours post-transfection of pEGFP-ITLN-1 into SGC-7901 cells, the ITLN-1 protein level was upregulated by 5.72 times （ P 〈 0. 01 ）. Meanwhile, the cellular proliferation and invasive activity were downregulated by52. 8% （P〈0.01） and 58. 1% （P〈0. 01）, respectively. Conclusion The expression vector of ITLN-1 was successfully constructed, and induced the ITLN-1 overexpression in gastric cancer cells, resulting in decrease of proliferation and invasion of cancer cells.

关 键 词：肠凝集素-1 胃癌 基因表达 

分 类 号：R735[医药卫生—肿瘤]