The Effect And Mechanism Of S100A13on Growth And Invasion Of Human Osteosarcoma Cells

Objective:To investigate the effect and mechanism of S100A13on growth and invasion of human osteosarcoma cells.Methods:1.The expression of S100A13mRNA in18cases of human osteosarcoma tissues and corresponding18cases of non-malignant normal bone tissues were examined by Real-time PCR;The expression of S100A13protein in8cases of human osteosarcoma tissues and corresponding8cases of non-malignant normal bone tissues were examined by Western-blot; The relationship between the expression level of S100A13mRNA and the clinical staging and prognosis of osteosarcoma were analysed.2. The expression level of S100A13protein in four human osteosarcoma cell lines(MG-63、U2-OS、HOS、Saos-2) and human osteoblastic cell line (hFOB1.19) were examined by Western blot, and the maximum expression cell line was selected for for subsequent experiments; The efficiency of transfection and interference were evaluated after siRNA targeting S100A13gene sequences by lentivirus transfection; Tumor cell growth curve experiments and MTT test were carried out to determine whether the interference the expression of S100A13in osteosarcoma cell affect the growth and proliferation of tumor cells; The scratch test were carried out to determine whether the interference the expression of S100A13in osteosarcoma cell affect the migration of tumor cells; Matrigel invasion assay were carried out to determine whether the interference the expression of S100A13in osteosarcoma cell affect the invasion of tumor cells; Tumor cell inoculation experiments in mice in vivo imaging to detect whether the interference the expression of S100A13in osteosarcoma cell affect the ability of tumorigenic.3.Western-blot and ELISA assay were used to examine the secretion of FGF-1in the culture supernatant after downregulating the expression of S100A13in osteosarcoma cell; Western-blot was used to examine the activation of downstream MAPK signaling pathways in osteosarcoma cells.Results:1.The expression of S100A13mRNA and protein in osteosarcoma tissue were significantly increased than that in normal tissue; The expression of S100A13mRNA in stage II were significantly increased than that in stage I; The S100A13high expression group was associated with better disease-free survival and overall survival time than S100A13low expression group.2.The expression level of S100A13protein in four human osteosarcoma cell lines(MG-63、U2-OS、HOS、Saos-2) were increased compared with the human osteoblastic cell line (hFOB1.19); Among the four osteosarcoma cell lines, the expression level of S100A13protein in the U2-OS cell line is the highest; The efficiency of transfection and interference were good in osteosarcoma cell by lentivirus transfection; The growth of osteosarcoma cell were dramatically inhibited after si-S100A13transfection compared with the scramble group;The proliferation of osteosarcoma cell were dramatically inhibited from the after si-S100A13transfection compared with the scramble group; The scratch test showed that the migration distance of osteosarcoma cell in the si-S100A13group were decreased compared with the scramble group; The number of invasion of osteosarcoma cell in the si-S100A13group were decreased compared with the scramle group; Tumor cell inoculation experiments in mice in vivo imaging showed that the ability of tumorigenic malignant behavior were inhibited after si-S100A13transfection.3.The FGF-1secretion in osteosarcoma cell showed significantly decreased in the si-S100A13group compared with the scramble group; The phosphorylation of FGFR-1、FRS2、c-raf、MEK1/2and Erk1/2(P-FGFR、P-FRS2、P-c-raf、P-MEK1/2and P-Erk1/2)showed decreased in the si-S100A13group compared with scramble group, whereas the total protein expression of FGFR-1、FRS2、MEK1/2and Erk1/2have no difference.Conclusion:1.The expression of S100A13mRNA and protein in human osteosarcoma tissue were significantly higher than that in normal tissue; The expression level of S100A13mRNA correlated with the clinical staging and prognosis of osteosarcoma.2.The downregulated the expression of S100A13in osteosarcoma cell inhibit the growth and proliferation of tumor cells, invasion and metastasis of tumors in vitro, and inhibit the ability of these tumorigenic malignant behavior in vivo;3.S100A13facilitate the FGF-1secretion and activation of downstream MPAK signaling pathways to promote cell proliferation and growth, invasion and metastasis, and tumor formation ability.