The Effect Of Noscapine Combination Of Cisplatin On Inhibition Proliferation And Apoptosis Of Human Osteosarcoma Cell Line MG-63

Background:Osteosarcoma (OS) is the most common primary bone tumor in children and young adults. With current therapies of OS, such as extensive surgical excision, radiotherapy and neoadjuvant chemotherapy, five-year survival rate has increased up to 50%-60%. Platinum-based drugs, such as cisplatin, are an important class of the most active chemotherapeutic agents that are widely used to OS. Cisplatin cytotoxicity involves the formation of intra and interstrand DNA adducts, and the resulting DNA damage triggers apoptosis.Noscapine has been primarily used as antitussive agent for several decades. Recent researches found that noscapine and its derivatives cause growth arrest of tumor cells in mitosis and induce apoptosis of tumor cells in vitro by interfering tubulin conformation and microtubule dynamics. As a matter of facts that noscapine and its derivatives show effectiveness against the growth of several tumor cells and little or no toxicity to human body, they have the potential to be an effective chemotherapeuticagent for the treatment of human cancers. However, the detailed mechanism of anticancer effect of noscapine at the molecular level was not exactly defined. This question, which is the mian purpose of present experiment, attracted our interest.Objectives:Noscapine, a phthalideisoquinoline alkaloid derived from opium, it has been primarily used as antitussive agent for several decades. Current researches indicate that it may also be one of the effective tumor suppressive agents. Hence, this present study was carried out to investigate the proliferation inhibition and apoptosis induction effect of noscapine on human osteosarcoma MG-63 cancer cells and the possible mechanisms.Methods:The MG-63 cells were treated with noscapine at various concentrations combination of cisplatin in present study. The MTT assay was used to assess cytostatic activity induced by noscapine and cisplatin. Flow cytometry were adopted to evaluate the cell apoptosis after noscapine treatment. Western blotting was used to demonstrate expression levels of involved protein such as Bcl-2, Bax, caspase-3.All the detection items in this study were repeated at least 3 times. Statistical analysis was done using SPSS software. The data was expressed as mean±SD and the statistical significance of the differences between control and noscapine treated cells was determined by a two-tailed Student's t test. The Spearman correlation test was used to analyze the correlation between reagents' concentrations and inhibition rates. P-value <0.05 was considered as significant.Results:The MTT assay indicated that the MG-63 cells proliferation could be dose-dependently inhibited by noscapine. Flow cytometry showed that noscapine significantly induced apoptosis in MG-63 cell line and this phenomenon was confirmed by DNA fragmentation analysis, As the cells displayed disintegrated nuclei and nonrandom DNA fragmentation. According to the results of Western Blot, bcl-2 and survivin were significantly downregulated in cells treated with the combination of noscapine and ciaplatin. Moreover, the different effects expressions of caspase-3 and PARP were detected by Western blot analysis. P-c-jun were upregulated,and p-erk were downregulated.Conclusions:In this study, the apoptosis induced effect of combination of celecoxib and cisplatin on human ostosarcoma MG-63 cell line was analyzed. The noscapine can inhibit growth, also can induce apoptosis of MG-63 cells. The findings presented here reveal that noscapine can induced marked apoptosis mainly by influencing JNK pathway and several Bcl-2 family members especially the balance of Bcl-2/Bax, which would trigger caspases-induced apoptosis. These data provide preclinical support for chemotherapeutic approach with noscapine for the treatment of ostosarcoma. All these results indicate that noscapine may represent a novel class of chemotherapeutic agents for ostosarcoma..