Expression and transfection of human papillomavirus 16E7 gene in human laryngeal carcinoma HEp-2 cell line
- VernacularTitle:人乳头状瘤病毒16E7在人喉癌细胞系中的转染及表达
- Author:
Jie QIU
- Publication Type:Journal Article
- Keywords:
papillomavirus,human;
laryngeal neoplasms;
transfection;
gene expression
- From:
Academic Journal of Second Military Medical University
1999;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct an eukaryotic expression vector harboring human papillomavirus 16E7(HPV16E7),and to study the effect of HPV16E7 on the biological character of human laryngeal carcinoma cell line HEp-2.Methods: The full sequence of human papillomavirus 16E7 gene was amplified from the plasmid of pET28a-HPV16E7 and was then cloned into pcDNA3.1/myc-His(-)B.The new plasmid,pcDNA3.1/myc-His-HPV16E7,was identified by sequencing and restriction digestion and was introduced into the laryngeal carcinoma cell line HEp-2 by electroporation;the morphology of the transfected cells was observed by the phase contrast microscope and the cell cycle was studied by flow cytometry.Western blot analysis was used to detect the expression of HPV16E7 proteins.Results: The plasmid,pcDNA3.1/myc-His-HPV16E7,was successfully constructed and was introduced into HEp-2 cells.The transfected cells proliferated actively with a prolonged S phase,and HPV16E7 proteins were detected in them.Conclusion: HPV16E7 can increase the tumorigenetic activity of HEp-2 cells.The construction of recombinant pcDNA3.1/myc-His-HPV16E7 paves a way for further study of laryngeal carcinoma.
