| Osteosarcoma is the most common primary tumor of bone tissue.It is more common and more malignant in young people.Currently,the treatment of osteosarcoma is mainly surgical and adjuvant chemotherapy.The formation and development of tumor is a multi factor,multi stage and gradual process.Early diagnosis and timely intervention are essential to the prognosis of the patients.In recent years,the study of cell proliferation and cell proliferation,cell signal transduction and cell signal has become a hot spot.With the development of oncology research,gene targeting therapy is possible,and it has brought hope for the attack of osteosarcoma.Gene targeting therapy is a molecular biological intervention to interfere in the development process of osteosarcoma,and to inhibit the malignant biological characteristics of tumors,so as to achieve the purpose of curing osteosarcoma.Therefore,the exploration and discovery of the new molecular targets for the treatment of osteosarcoma will provide data support and theoretical basis for gene targeting therapy.The latest research has found that nuclear receptorco-activator 5 protein(NCOA5)is a nuclear receptor co regulation factor,which has 579 amino acids that are widely expressed.It is also an estrogen receptor and nuclear receptor coactivator.NCOA5 is a kind of estrogen that mediates estrogen in human cells.NCOA5 can also regulate gene defects,which can lead to cancer and diabetes.Curcumin is a kind of natural plant polyphenols,which is extracted from the turmeric of the ginger plant and also in other ginger plants.It is widely found in the rhizomes of the Chinese traditional Chinese medicine curcuma.It has been reported that curcumin can inhibit invasion and metastasis of tumor cells.In curcumin,whether curcumin can prevent tumor invasion and metastasis through NCOA5 target has not been reported clearly.The effect of curcumin with different concentrations on the expression of NCOA5 in human osteosarcoma cell line MG-63 was studied,and the mechanism of invasion and metastasis of osteosarcoma was further explored,providing a theoretical basis for curcumin in the clinical treatment of osteosarcoma.This study explored and studied the role of NCOA5 in the development of osteosarcoma and the mechanism of curcumin inhibiting the invasion of osteosarcoma MG-63.It has important scientific significance for further exploration of new molecular therapeutic targets of osteosarcoma.Part I Study on the relationship between NCOA5 expression and postoperative prognosis of osteosarcoma patientsTo investigate the relationship between the expression of nuclear receptorco-activator5 protein(NCOA5)and the prognosis of postoperative patients with osteosarcoma.Human osteosarcoma were collected from 145 patients and normal bone tissues were collected from 100 individuals as controls.Immunohistochemistry technique(IHC)and reversed transcriptast-polymerase chain reaction(RT-PCR)were employed to measure nuclear receptorco-activator 5 protein in cases of human osteosarcoma.IHC staining showed that osteosarcoma nuclear receptorco-activator 5 protein positive rate were 26.21%.The positive rate of normal bone tissues were 82.00%,which were significantly higher than that of human osteosarcoma tissues.RT-PCR showed that the positive rates of NCOA5 m RNA expression in human osteosarcoma were 17.24%.The positive rate of normal bone tissues were 84.00%,which were significantly higher than that of human osteosarcoma tissues.The survival of patients with osteosarcoma not only was related to the expression of NCOA5 protein,but also related to the tumor location and differentiation and staging.We have come to the conclusion that NCOA5 m RNA and protein expression are consistent in human osteosarcoma tissues.The expression of NCOA5 is low in human osteosarcoma tissues,while its expression is high in normal bone tissues.The low expression of NCOA5 is closely related to the human osteosarcoma.which was aslo associated with postoperative survival of humanosteosarcoma.Part II Study on the mechanism of curcumin on NCOA5 gene and MG-63 cellTo investigate the regulatory effect of curcumin on expression of NCOA5 in human osteosarcoma as well as the possible mechanism of curcumin in prevention and treatment of human osteosarcoma.Highly invasive MG-63 cells were treated with curcumin at various dosages.The cytotoxic effects of treatment with 5,10,15,20,25,30,35,40 and 50umol/L curcumin for 24,48 and 72 h on MG-63 cells were measured by MTT assay.The treatment with curcumin at a dosage of more than 15 umol/L for more than 24 h inhibited the growth of MG-63 cells in time-and dose-dependent patterns.The treatment with curcumin at a dosage of not less than 15 umol/L for 24 h showed no significant cytotoxic effect on the cells,while the survival rate of cells was more than 85%.The invasion abilities of cells treated with 5,10 and 15 umol/L curcumin were measured by Transwell test,while the adhesive abilities by cell adhesion assay.The invasion and adhesive abilities of cells decreased gradually with the increasing curcumin concentration.The curcumin at a concentration of 15 umol/L showed the strongest inhibitory effect on the two abilities.The effect of 5,10 and 15 umol/L curcumas on expression levels of NCOA5 were determined by western blot,while those on transcription level of NCOA5 m RNA by RT-PCR.Curcumin showed significantly dose-dependent inhibitory effect on the transcription level of NCOA5 m RNA.We have come to the conclusion that curcumin inhibited the invasive abilitiy of MG-63 cells by inhibiting the activation of expression of NCOA5 as a target gene of the pathway.To sum up,the expression of NCOA5 plays an important role in the development of osteosarcoma.Curcumin can promote the expression of NCOA5 and inhibit the invasiveness of MG-63 cells,thus inhibiting the occurrence and development of osteosarcoma.And it also can be used as a potential molecular target for the treatment of osteosarcoma. |
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