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Mechanism of Podophyllotoxin rheinate (RH-01) on human osteoma sarcomatosum cells growth

WAN Zong-ming, CHEN Hong, CAO Bo, WANG Ying, MOU Hong-jun(Department of Pharmacognosy, Medical College of Chinese Peoples Armed Police Forces, Tianjin 300162, China)  
Objective To study the effects of Podophyllotoxin rheinate (RH-01) on human osteoma sarcomatosum (HOS) cells. Methods HOS Cells were cultured in MEM alone or exposed to different concentrations of RH-01. The growth inhibition was analyzed by MTT assay. The bone-targeted effect of RH-01 was evaluated by values of hydroxyapatite ceramic adsorption. The flow cytometry (FCM), fluorescence microscopy, and RT-PCR assay were employed to detect the action mechanism of RH-01. Results RH-01 Remarkably inhibited HOS cells growth, its IC50 value was 0.18 μmol/L at 72 h. Values of hydroxyapatite ceramic adsorption towards AcM and RH-01 were (8.1±0.15) and (14.8±0.11) μmol/g, respectively, which demonstrated that RH-01 had much stronger bone-targeted function than AcM. After the HOS cells with RH-01 diluted by Hoechst 33342 solution, it showed typically morphologic change compared with the control group under fluorescence microscope. Results indicated that RH-01 dramatically induced an accumulation in G2/M phase of cell cycle at 24 h by FCM analysis. RT-PCR Assay showed that the expression of cyclin D1, CDK4 mRNA obviously decreased in a concentration dependent manner. Conclusion The compound RH-01 could obviously inhibit the growth of HOS cell line and have a very strong bone-afinity feature in vitro.
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