Establishment of ovarian SKOV3 cell line stably expressing PES1 by Tet-on inducible system
10.3760/cma.j.issn.1673-422X.2012.06.020
- VernacularTitle:利用Tet-on可诱导系统构建稳定表达PES1的SKOV3细胞株
- Author:
Jieping LI
;
Qinren ZHUANG
;
Xiaopeng LAN
;
Guobin ZENG
;
Xiaofeng LUO
- Publication Type:Journal Article
- Keywords:
Gene expression;
Ovarian neoplasms
- From:
Journal of International Oncology
2012;39(6):465-468
- CountryChina
- Language:Chinese
-
Abstract:
Objective To further research the biological functions of PES1,the ovarian SKOV3 cell line with inducible stable PES1 expression is established by using Tet-on system.Methods PES1 was cloned into pTRE-Tight vector via PCR and its expression was identified. After transfected the regulating plasmid pTet-on,SKOV3 cells were screened with G418 and re-transfected pTRE-Tight-PES1.The positive cell clones were screened out with hygromycin and were induced by doxycycline (Dox) to definite the best induction concentration.Growth velocity of SKOV3 cells stably expressing PES1 induced by Dox was detected with viola crystallina.Results The SKOV3 cells with inducible PES1 expression were screened out after the cells were transfected pTRE-Tight-PES1 constructed.Dox could dose-dependently induce the PES1 expression with the concentration under 2 mg/L,and 2 mg/L of Dox induced the highest PES1 expression.Growth velocity of SKOV3 cells transfected pTRE-Tight has no significant difference between the SKOV3 cells transfected nothing induced with Dox.However,the SKOV3 cells transfected pTRE-Tight-PES1 grew faster than the cells transfected pTRE-Tight or without transfection in the fourth day (P =0.001 ).Conclusion The inducible stable PES1 expression SKOV3 cells are successfully established and could be used to be an effective cell model to research the biological functions of PES1.The expression of PES1 could promote the growth of SKOV3 cells.
