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岭南现代临床外科 ›› 2019, Vol. 19 ›› Issue (01): 44-47.DOI: 10.3969/j.issn.1009-976X.2019.01.009

• 论著与临床研究 • 上一篇    下一篇

质粒介导LAPTM4B过表达胰腺癌细胞株PANC?1的构建及鉴定

张光涛1，梁悦2，李明华3   

  1. 北京大学深圳医院1.肝胆胰外科；2.药学部；3.医学中心，广东深圳 518036
  • 通讯作者: 张光涛

Construction and verification of plasmid mediated LAPTM4B over expressing pancreatic cancer cell line PANC?1

ZHANG Guangtao1， LIANG Yue2， LI Minghua3   

  1. 1. Hepatobiliary Pancreatic Surgery，Peking university Shenzhen Hospital，Shenzhen 518030，China； 2. Pharmacy Department， Peking University Shenzhen Hospital，Shenzhen 518030，China；3.Peking University Shenzhen Hospital， Shenzhen 518030， China.
  • Online:2019-02-20 Published:2019-02-20

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摘要： ［摘要］ 目的 筛选低表达LAPTM4B?35细胞株并构建质粒介导的LAPTM4B过表达胰腺癌细胞株并进行鉴定。方法 通过RT?PCR及Western?Blot 方法检测3种胰腺癌细胞LAPTM4B的表达，筛选出现低表达LAPTM4B细胞株。通过质粒稳定转染胰腺癌细胞PANC?1，pcDNA3.0?AE过表达LAPTM4B基因，应用RT?PCR，Western?blot进行鉴定。结果 成功扩增提取pcDNA3.0?AE质粒，转染并筛选稳定细胞株。RT?PCR及Western?Blot 显示相对空白对照，稳定株的过表达LAPTM4B?35明显。结论 成功构建筛选低表达细胞株，并建立稳定过表达 LAPTM4B的胰腺癌细胞株。

关键词: PANC?1, LAPTM4B, 质粒, 稳定株, 胰腺癌

Abstract: ［Abstract］ Objective To select low expression LAPTM4B?35 cell lines and construct and verify the stable PANC?1 cell line infected with LAPTM4B over?expressing plasmid. Methods Via RT?PCR and western?Blot， the expression of LAPTM4B in three kinds of pancreatic cancer cells was detected. The low expression LAPTM4B cell lines were screened. Overexpression of LAPTM4B?35 in PANC?1 cells by plasmid stable transfection was identified by RT?PCR and Western?blot. Results PcDNA3.0?AE plasmid was successfully amplified and transfected， and stable cell lines were screened. RT?PCR and western?blot showed that the overexpression of LAPTM4B?35 in stable strains was significantly higher than that of the blank control group. Conclusion The low expression cell line was successfully screened and the stable cell line PANC?1 overexpressing LAPTM4B?35 was established.

Key words: stable cell line, plasmid, pancreatic cancer, PANC?1, LAPTM4B?35

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