The Study On The Expression Of KiSS-1 And GPR54 Gene In Osteosarcoma

[Objective] The aim of this study is to evaluate the mRNA and protein expression levels of KiSS-1 and GPR54 gene in osteosarcoma cell lines and tissue specimen, explore the relationship between the expression of KiSS-1 or GPR54 gene and the biologic behavior and prognosis of ostosarcoma, and further analyze the relationship between KiSS-1 gene and GPR54 gene in osteosarcoma.[Methods] 1.The osteosarcoma cell lines MG-63, Saos-2, U-2 OS and osteoblast cell line hF-OB1.19 are cultured in. 2. Expression levels of KiSS-1 and GPR54 gene mRNA in 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS) and normal osteoblast cell line (hF-OB 1.19) were examined using semi-quantitative RT-PCR and immunochemistry staining. 3. Expression levels of the protein products of KiSS-1 and GPR54 gene in 3 types of osteosarcoma cell lines (MG-63, Saos-2 and U-2 OS) and normal osteoblast cell line (hF-OB 1.19) were examined using immunochemistry staining. 4. Transwell assay was used to detect the invasion ability of 3 types of osteosarcoma cell lines. 5. Immunochemistry staining was used to examin the expression levels of the protein products of KiSS-1 and GPR54 gene in 44 cases of osteosarcoma and 12 cases of osteochondroma paraffin specimens.[Results] 1. The expression level of KiSS-1 mRNA and protein in normal osteoblast cell line hF-OB1.19 is significant higher than in 3 types of osteosarcoma cell lines. With the malignant degree increased, the expression level of KiSS-1 mRNA and protein decreased in the 3 types of osteosarcoma cell lines. 2. Transwell assays displayed a gradually increased aggressive phenomenon in osteosarcoma cell lines U-2 OS, Saos-2 and MG-63. 3. Patients with KiSS-1 protein positive expression had a shorter median metastasis time than patients with KiSS-1 protein negative expression, and the difference was significant. 4. Patients with KiSS-1 protein positive expression had a shorter median survival time than patients with KiSS-1 protein negative expression, and the difference was significant. 5. GPR54 mRNA and protein expressed strongly in all the 3 types of osteosarcoma cell lines and normal osteoblast cell line hF-OB 1.19, but no significant difference existed between each other. 6. Patients with GPR54 protein positive expression had a shorter median survival time than patients with GPR54 protein negative expression, and the difference was significant. 7. Patients with both KiSS-1and GPR54 protein positive expression had a higher mortality rate than patients with both KiSS-1and GPR54 protein negative expression, and the difference was significant.[Conclusions] 1. The expression level of KiSS-1 mRNA and protein in normal osteoblast cell is significant higher than in 3 types of osteosarcoma cell. With the gradually increased aggressive ability, the expression level of KiSS-1 mRNA and protein decreased in the 3 types of osteosarcoma cell lines, which indicate KiSS-1 gene maybe play a role of supression of metastasis in osteosarcoma cell lines. 2. The expression level of KiSS-1 is higher in osteosarcoma patients than in benign osteochondroma patients, the expression level of KiSS-1 is positve correlation with the metastasis and negative correlation with the survival time in osteosarcoma patients. 3. GPR54 mRNA and protein expressed strongly in all the 3 types of osteosarcoma cell lines and normal osteoblast cell line hF-OB 1.19, but no significant difference existed between each other. 4. The expression level of GPR54 is higher in osteosarcoma patients than in benign osteochondroma patients, and the expression level of GPR54 is negative correlation with the survival time in osteosarcoma patients. 5. Both KiSS-1and GPR54 protein positive expression is negative correlation with the survival time in osteosarcoma patients.