Inhibitory effects of recombinant adnovirus carrying wild type PTEN gene on endometrial carcinoma cells: an in vitro study / 第二军医大学学报

ABSTRACT

Objective:To observe the expression of Ad-PTEN in human endometrial carcinoma cell line RL95-2 after in vitro infection and investigate the mechanism by which Ad-PTEN inhibits tumor cell proliferation and induces apoptosis. Methods: Ad-PTEN was constructed through a bacterial homologous recombinant system; the expression of Ad-PTEN in RL95-2 cells was determined by RT-PCR, Western blotting and immunohistochemical staining. The efficiency of adenovirus mediated gene transfer of Ad-PTEN was determined by X-gal staining. The inhibitive effect of Ad-PTEN on RL95-2 cells proliferation was determined by cell growth analysis and MTT assay; the morphologic and ultrastructural changes of RL95-2 cells transfected with Ad-PTEN were observed by the light and electron microscopy; and Flow cytometry was used to study the cell cycle and apoptosis. Results: The expression of Ad-PTEN mRNA and protein in RL95-2 cells was confirmed by RT-PCR, Western blot and cell immunohistochemical staining. The efficiency of adenovirus mediated Ad-PTEN gene transfer was 100% when the multiplicities of infection (MOI) was 50. Exogenous PTEN gene significantly suppressed the growth of RL95-2 cells and iduced the apoptosis. Ad-PTEN could also induce cell cycle arrest (G_(0)/G_(1) ) and activated caspase-3 . Conclusion: The constructed Ad-PTEN transfection system is highly efficient in introducing wild type PTEN gene into human endometrial carcinoma RL95-2 cells. Ad-PTEN can strongly inhibit cell proliferation and induce apoptosis in RL95-2 cells, which may be associated with cell cycle arrest (G0/G_(1)) and the activation of caspase-3.