Study On The Apoptosis Of Acute Myeloid Leukemia SHI-1Cells Induced By Puerariae Radix Flavones

[Backgroup and Objective]Puerarin is one of the main compositions of Xue-Fu-kang, which is sourced from the NO.1hospital affiated to Nanjing Univesity of Chinese Medicine, creating in1989based on disintoxicating and blood flow promoting,could effectively in cure of chronic myeloid leukemia and acut myeloid leukemia. Rencently years research team experiments results manifested PRF with concentration of50-200μg/ml could selectively inhibit the proliferation of differate cell lines of AML.in the study, we used SHI-1cells which have MLL-AF6leukemiagenesis fusion gen and p53gene abornomal as the objiect to explore the effect of PRF on proliferation and apoptosis in SHI-1and explore its possible mechanism.[Methods]1)0-200μg/ml PRF treate SHI-1cells for24,48and72hours, the cells proliferation inhibitory rates were analyzed by MTT assay, and chosed profectable concentration.2)0、50、100、150、μg/ml PRF treat SHI-1cells for48hours, the morphology changes were detected microscopically by Wright’s-Giemsa staining, cells early-stage apoptotic rates were detected by FITC-AnnexinⅤ/PI double staining with flow cytometry(FCM), cells cycle were detected by FCM.3)0、50、100、150μg/ml PRF treat SHI-1cells for48hours, MLL-AF6were detected by RT-PCR.[Results]MTT assay shows PRF (0-200μg/ml) can significantly inhibited the proliferation of SHI-1cell, the inhibiting effect conformed to time and dose-dependence, by spass software analyze SHI-150%inhibiting concentration is136.87μg/ml at48h. After being treated with PRF (0、50、100、150150μg/ml) for48h, the classical morphologic characteristics of apoptosis was demonstrated under the light microscope. FITC-Annexinv/PI show the eariy apoptosis of the SHI-1cells was elevated, which as dose-dependent manner. Cell cycle analyses showed that cells in G1phase decreased, while S phase increased as the dose of PRF increased, cells were arrested in S-phase. RT-PCR demonstrate as PRF concentration increased, MLL-AF6has no significantly changes.[Conclusion]PRF (≤200μg/ml) might inhibit SHI-1cells proliferation in vitro in time and...