Zhang Chun-hua, Wen Ze-qing, Li Jian-feng, Li Chang-zhong, Shi Min, Yang Gui-wen, Lan Shou-min, Zhu Yong, Wang Fei, Zhang Yao-jing, Wang Ying-ying, Zhang Hui
Department of Obstetric and Gynecology, Shandong University Shandong Provincial Hospital, Jinan, Shandong 250021, China.
Chin Med J (Engl). 2008 Jan 20;121(2):166-71.
Rosiglitazone is known as the most potent and specific peroxisome proliferators-activated receptor gamma (PPAR-gamma) 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 beta3 (TGF-beta3) and the cell proliferation in human uterine leiomyoma cells in vitro.
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-gamma and TGF-beta3. Immunofluorescence staining was used to detect the expressions of PPAR-gamma and TGF-beta3 proteins.
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-gamma mRNA expression was up-regulated and TGF-beta3 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-gamma and TGF-beta3 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.
The present study demonstrates that the PPAR-gamma activator, rosiglitazone, inhibits the cell proliferation partly through the regulations of PPAR-gamma and TGF-beta3 expressions. The cross-talk between the signal pathways of PPAR-gamma and TGF-beta3 may be involved in the process.
罗格列酮是已知最强效且最具特异性的过氧化物酶体增殖物激活受体γ（PPAR-γ）配体。它对从癌症到动脉粥样硬化和糖尿病等病理生理过程可能具有深远影响。然而，罗格列酮是否影响体外培养的人子宫平滑肌瘤细胞中转化生长因子β3（TGF-β3）的蛋白表达及细胞增殖尚不清楚。
解剖并培养人子宫平滑肌瘤组织。细胞分为5组：1个对照组和其他4个不同浓度罗格列酮组（10⁻⁷、10⁻⁸、10⁻⁹和10⁻¹⁰mol/L）。细胞在无血清的杜尔贝科改良伊格尔培养基中培养72小时。采用MTT比色法检测细胞增殖。逆转录聚合酶链反应（RT-PCR）用于检测PPAR-γ和TGF-β3的mRNA表达。免疫荧光染色用于检测PPAR-γ和TGF-β3蛋白的表达。
MTT比色法表明，罗格列酮处理（10⁻⁷至10⁻⁹mol/L）72小时后导致细胞生长受到抑制（P < 0.01）。RT-PCR分析显示，10⁻⁷mol/L罗格列酮在72小时时显著影响基因表达：PPAR-γ mRNA表达上调，TGF-β3 mRNA表达下调，且10⁻⁷mol/L的罗格列酮对此影响最为有效（P < 0.01）。免疫荧光染色表明，与其他处理组和对照组相比，10⁻⁷mol/L罗格列酮处理72小时后导致PPAR-γ和TGF-β3蛋白表达发生显著变化（P < 0.01）。罗格列酮对子宫平滑肌瘤细胞的所有作用在体外均呈剂量和时间依赖性。
本研究表明，PPAR-γ激活剂罗格列酮部分通过调节PPAR-γ和TGF-β3的表达来抑制细胞增殖。PPAR-γ和TGF-β3信号通路之间的相互作用可能参与了这一过程。
