Effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line
- VernacularTitle:反义MBD1基因真核表达载体转染对人胆管癌细胞MBD1基因表达的影响
- Author:
Shi ZUO
;
Zhanfei LI
;
Jian LUO
;
Wei GUO
;
Lining XU
;
Minfeng LIU
;
Jingqing DONG
;
Shengquan ZOU
- Publication Type:Journal Article
- Keywords:
DNA,Antisense;
Bile Duct Neoplasms/etiology;
Transfection;
Gene Expression
- From:
Chinese Journal of General Surgery
2000;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the effect of transfection of antisense MBD1 gene eukaryotic expression vector on the expression of MBD1 gene in human cholangiocarcinoma cell line QBC-939.Methods The(constructed) antisense MBD1 gene eukaryotic expression vector was transfected into the human(cholangiocarcinoma) cell line QBC-939 using lipofectamine transfection reagents,and positive cell clones were obtained using G418 selection after transfection.The constructed recombinant vector was transfected into(QBC-939) cells successfully and was confirmed by amplifying the exogenous neo~R gene with PCR method.The expression level of MBD1 gene mRNA and protein was detected by RT-PCR and FCM methods respectively.Results Following the transfection,the MBD1 gene mRNA level in human cholangiocarcinoma cell line QBC-939 decreased from 0.912?0.022 to 0.215?0.017,and the MBD1 gene protein level also(decreased) from(80.19?5.05)% to(35.11?4.05)%.There were very significant differences on the expression both at the transcription and post-transcription levels of MBD1 gene between non-tranfection group and the antisense MBD1 gene eukaryotic expression vector transfection group(P
