Inhibition of proliferation and transforming growth factor beta3 protein expression by peroxisome proliferators-activated receptor gamma ligands in human uterine leiomyoma cells
Zhang, C-hua.; Wen, Z-qing.; Li, J-feng.; Li, C-zhong.; Shi, M.; Yang, G-wen.; Lan, S-min.; Zhu, Y.; Wang, F.; Zhang, Y-jing.; Wang, Y-ying.; Zhang, H.
Chinese Medical Journal 121(2): 166-171
2008
ISSN/ISBN: 0366-6999 PMID: 18272045 Document Number: 622591
Background Rosiglitazone is known as the most potent and specific peroxisome proliferators-activated receptor y (PPAR-y) ligand. It has potentially far-reaching effects on pathophysiological processes, from cancer to atherosclerosis and diabetes. However, it is not clear whether rosiglitazone affects the protein expression of transforming growth factor 133 (TGF-beta 3) and the cell proliferation in human uterine leiomyoma cells in vitro.Methods Human uterine leiomyoma tissues were dissected and cultured. Cells were divided into 5 groups: one control group and other four groups with different concentrations of rosiglitazone (10(-7), 10(-8), 10(-9) and 10(-10) mol/L). Cells were cultured for 72 hours in serum-free Dulbecco's modified Eagle's medium. MTT reduction assay was used to detect the cell proliferation. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of PPAR-y and TGF-beta 3. Immunofluorescence staining was used to detect the expressions of PPAR-y and TGF-beta 3 proteins.Results MTT reduction assay indicated that the treatment with rosiglitazone (from 10-7 to 10-9 mol/L) resulted in an inhibition of the cell growths after 72 hours (P<0.01). RT-PCR analysis revealed that 10-7 mol/L rosiglitazone significantly affected the gene expression at 72-hour: PPAR-y mRNA expression was up-regulated and TGF-beta 3 mRNA was down-regulated and rosiglitazone at the concentration of 10-7 mol/L affected these most effectively (P<0.01). Immunofluorescence staining demonstrated that treatment with 10-7 mol/L rosiglitazone resulted in the significant changes of PPAR-y and TGF-beta 3 protein expressions compared with the other treatment groups and the control group at 72-hour (P<0.01). All the effects of rosiglitazone on uterine leiomyoma cells were dose- and time-dependent in vitro.Conclusions The present study demonstrates that the PPAR-y activator, rosiglitazone, inhibits the cell proliferation partly through the regulations of PPAR-y and TGF-beta 3 expressions. The cross-talk between the signal pathways of PPAR-y and TGF-beta 3 may be involved in the process.