| Background and Objectives:Osteosarcoma is one of the malignant tumors with high mortality among adolescents in the world,and the mortality rate is about 60% of the juvenile malignant tumor.In recent years,with the use and improvement of surgery combined with chemotherapy,the 5-year survival rate has improved,but still less than 60%.Osteosarcoma cells are frequently associated with tumor after mutation gene expression changes in gene expression,exploring the molecular mechanism and its malignant potential is to control bone intrusive tumor invasive growth,to prevent the recurrence of the key.CHI3L1 gene is found in human chromosome 1q31-q32,and the gene group leader is about 10.9kb,with 10 exons.Most tissues of human body,such as fat and bone,have CH13L1 gene.Chitinase like protein(YKL-40)is a product its genetic code,research reports in recent years,a variety of tumor proliferation,differentiation,infiltration,in the process of angiogenesis and invasion and metastasis are the proteins involved in.In recent years about the malignant degree of osteosarcoma between high tumor related research may have all the time,the research CHI3L1 gene expression in osteosarcoma related content rarely,and have their own confine.And this study was to further explore expression of the CHI3L1 gene and what role it plays in osteosarcoma,looking for a new index for osteosarcoma condition assessment,as well as the maybe molecular target therapy and gene therapy in the future provide the basis and the reference.In this study,the main purpose of this study was to observe the expression of CHI3L1 gene in osteosarcoma cells,and then to improve the expression of CHI3L1 gene in osteosarcoma cells.The first part of the experiment was to detect the expression of gene in human osteosarcoma.The second part was to detect the differences in mRNA and protein expression of CHI3L1 in human osteosarcoma cell line MG63,U20 S and human osteoblasts hFOB1.19.The third part is to compare the differences between the osteosarcoma cell model and the normal osteosarcoma cell in vitro and in vitro apoptosis by constructing a model of osteosarcoma of CHI3L1 gene.Methods:In the first part of the experiment,three patients with osteosarcoma were randomly selected and the total RNA of normal cancellous bone tissue was extracted from the same patient,and the difference was found by gene chip analysis(CHI3L1).Then,the expression of 21 cases of osteosarcoma and the adjacent tissue was further identified by real-time fluorescence quantitative PCR and fluorescence in situ hybridization.The second part of the experiment: the bone sarcoma cell line MG63,U2 OS cell lines hFOB1.19 and people development as the research object,using rt-pcr and Western immunoblot method to detect different cell lines in the mRNA and protein expression of CHI3L1 gene.Transwell experiment,CCK8 cell count experiment and flow cytological analysis were used to detect the proliferation activity and cell cycle of MG63 and U2 OS after intervention of CHI3L1.Results:1.The results showed that the expression of CHI3L1 in osteosarcoma decreased significantly.The results of real-time fluorescence quantitative PCR and fluorescence in situ hybridization showed that the expression of CHI3L1 in the 20 osteosarcomas was significantly higher than that of the tumor parenchyma.2.The expression of CHI3L1 gene in human osteosarcoma cell line MG63 and U2 OS increased,and corresponding protein expression was increased.3.Transfection pcDNA3.1/CHI3L1 fusion plasmid in MG63 and U2 OS can quicken cell cycle and significantly promote cell proliferation.Conclusion:1.The expression of CHI3L1 as a tumor suppressor gene was decreased in osteosarcoma.2.CHI3L1 is used as a tumor suppressor gene to express and function in osteosarcoma. |
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