Study Cytotoxicity Effect And Mechanism Of Matrine On Human Osteosarcoma Cell Line MG-63

Background Osteosarcoma is the most frequent primary bone malignancy affecting children and young adults, with the feature of growth fast, easy to metastasis and high degree of malignancy. The five-year survival rate for patients with osteosarcoma was estimated at 20% before twenty years. Recently, with current protocols including a combination of neoadjuvant chemotherapy and limb salvage surgery, the five-year disease free survival has increased to over 60%. Despite this substantial improvement, approximately 40% of patients will either present with metastasis at the time of diagnosis or develop metastasis over the course of the disease. Thus, further therapeutic improvement and research for new drugs sensitive to osteosarcoma cells may increase the survival rate. Matrine is the main effective component of the Sophora flavescens Ait. Recently, it has been demonstrated that matrine hasevident cytotoxicity against variety cancer cells including K562 cells, HepG2 cells, SMMC-7721 cells, et al. in vitro. The mechanism of cytotoxicity is that matrine can inhibit proliferation and induce apoptosis of cancer cells. Objective of our study is to observe the growth inhibitory and apoptosis-inducing effects of matrine on the human osteosarcoma cell line MG-63 in vitro.OBJECTIVE To study the cytotoxicity effect and mechanism of matrine on human osteosarcoma cell line MG-63.METHODS The proliferation effect of MG-63 cell line cultured with matrine in different concentrations and variable durations, at the same time was detected by MTT assay. Alteration of cell cycle of different concentration treating and control groups at the third day was analyzed by Flow cyctometry (FCM) . In addition, the apoptosis was detected by analyzing the alteration of cell DNA content. The morphological change of MG-63 cell cultured with matrine in different concentrations was investigated by electronic microscopy.RESULTS Matrine can inhibit proliferation of MG-63 cells. The effect of proliferation inhibition was more prominent with the treated time prolonging and the drug concentration increasing, which showed a time-dependent and dose-dependent manner.. Increased G1/G0 phase rate, decreased S phase rate and pre-G1 cells were showed by FCM after matrine-treated MG-63 cells. In addition, FCM analysis showed that therewere sub-G1 peaks(apoptosis peaks) in all treatment groups. In the control group and 0.25, 0.50, 0.75, 1.0, 1.25mg/ml treatment groups, the rate of apoptotic cells to total cells were 1.23%, 6.71%, 21.50%, 32.92%, 45.79%, 51.90%, respectively. In the experiment range, the apoptosis-inducing effect of matrine was related with drug concentration. Electron microscope revealed the characteristic apoptosis alterations, shrinking cellular morphology, integrity cell membrane, karyopycnosis, fragment of nucleus, chromatin condensation and margination, crescent nucleus, cytoplasmic vacuoles.CONCLUSION Matrine has cytotoxicity against human osteosarcoma cell line MG-63. The mechanism includes that matrine can effectively inhibit the proliferation of MG-63 cell line, arrest the cell cycle at G0/G1 phase and induce the apoptosis of MG-63 cell line. The effect of matrine displays a time-dependent and dose-dependent manner.