Cloning and analyzing the activity of the promoter of the MUC1 gene in Panc-1 cells
10.3760/cma.j.issn.1007-8118.2014.02.015
- VernacularTitle:MUC1黏蛋白启动子的克隆及在胰腺癌Panc-1细胞中的转录活性
- Author:
Yaozhen PAN
;
Chengyi SUN
;
Lei ZHAN
;
Hao ZHANG
;
She TIAN
;
Hong ZHANG
;
Nan JIANG
- Publication Type:Journal Article
- Keywords:
Pancreatic cancer;
MUC1 Promoter;
Dual luciferase assays
- From:
Chinese Journal of Hepatobiliary Surgery
2014;20(2):137-141
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the MUC1 promoter's role in driving gene expression in pancreatic cancer and its therapeutic significance.Methods Two plasmids were made.The plasmid pEGFP-MUC1N1 contained MUC1 promoter fragment connected to the pEGFP-N1 vector with the EGFG reporter gene.The pShuttle-MUC1-EGFP plasmid contained MUC1 promoter fragment and EGFP reporter gene connected to pShuttle plasmid.Lipofectamine 2000 was used to transfect the two plasmids into cells of MUC1-positive human pancreatic cell line Panc-1 and MUC1-negative human cervical carcinoma Hela.Fluorescence microscopy and flow cytometry compared the specificity and activity of the MUC1 promoter and CMV promoter.Results Reporter gene EGFP-positive cells 48 hours after transfection with pEGFP-MUC1-N1 and pShuttleMUC1-EGFP plasmid were 69.6％ and 63.6％ respectively,in Panc-1 cells,and 4.2％ and 3.7％ respectively,in Hela cells.Conclusions MUC1 promoter can drive reporter gene activity in MUC1-positive tumor cells targeting functional expression.There is potentially a use of targeted therapy in pancreatic cancer at the genetic level.
