The Investigation Of Rosiglitazone On The Growth Inhibition And Apoptosis Induction In Human Cervical Cancer Hela Cells

Objective: To investigate the rosiglitazone on induction of growth inhibition and apoptosis of human cervical cancer Hela cells line, and its mechanism in vitro.Methods: Human cervical cancer Hela cells were cultured in vitro. MTT assay was used to determine the effect of rosiglitazone on the viability of Hela cells. Fluorescence after AO/EB staining was used to observe the morphologic changes of apoptosis induced by rosiglitazone in Hela cells line. DNA agarose gel electrophoresis was used to test apotosis induced by rosiglitazone in Hela cells line. Flow cytometry (FCM) using PI staining was used to analyse the Sub-G1 cell population treated with rosiglitazone. COX-2 was determined using cell immunofluorescence.Results: In preliminary experiment, Treatment of Hela cells with various concentrations of 1.0, 10.0, 100.0, 200.0, 500.0μmol/L rosiglitazone for 24, 48, 72 hours, respectively. MTT assay showed that the best time is 48 hours, and efficacy of both 200.0, 500.0μmol/L of rosiglitazone for inhibition of viability in Hela cells was nearly equal, there was no significant. In subsequent experiments, we used the experimental concentration gradient 1.0, 10.0, 100.0μmol/L and incubated for 48 hours. The results shown that the inhibitory rateof relative ecell viability was (15.42±2.2)%, (54.34±2.78)%,(82.31±4.87)%, respectively, and its IC50 was 9.66μmol/L. Typical morphologic changes of apoptosis in Hela cells could be observed after treatment with rosiglitazone by fluorescence microscope using AO/EB fluorescence staining. DNA agarose gel electrophoresis shown that DNA ladder bands could appear after treatment with rosiglitazone at 100.0μmol / L for 48h. Flow cytometry(FCM) analysis after PI stainning indicated that the percentage of sub-G1 cell population in Hela cells was( 5.15±0.48 )%,(17.7±0.81)%,(38.9±0.98)% respectively after treatment with rosiglitazone at the concentrations of 1.0, 10.0, 100.0μmol/L for 48h. FITC fluorescence labeling immunocytochemistry showed treatment with 1.0, 10.0, 100.0μmol/L of rosiglitazone reduced positive cells for COX-2 protein expression to(39.59±0.32)%, (30.36±0.81)% , (20.12±0.46)%, in a concentration dependent manner.Conclusion:1) Rosiglitazone posses the inhibitory effect of the viability of Hela cells, in a dose-dependent manner.2) Rosiglitazone can significantly induce apoptosis of Hela cells, in a dose-dependent manner.3) Inhibition of cell viability and induction of apoptosis by rosiglitazone in human cervical cancer Hela cells may be associated with downregulation of intracellular COX-2 protein expression.