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The Effect of SPARC Gene Over-expression on Biological Characteristics of Human Ovarian Carcinoma Cells with Directional High Lymphatic Metastasis
Xie Yihong1, Zhong Yanping2, Wang Qi1, Li Li1, Zhang Xinying2, Ruan Heyun2, Li Danrong1*
1Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China;2Medical Scientific Research Centre of Guangxi Medical University, Nanning 530021, China
Abstract: The aim of this study was to investigate the effect of secreted protein acidic and rich in cysteine gene (SPARC) overexpression on biological characteristics of human ovarian carcinoma cells with directional high lymphatic metastasis (SKOV3-PM4). SPARC gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into SKOV3-PM4 cells to upregulate the expression of SPARC gene. The expression level of SPARC was detected at both mRNA and protein levels by Real-time PCR and Western blot. The protein distribution was observed by cell immunofluorescence under laser confocal microscopy. The proliferative ability of SKOV3-PM4 cells was detected by cell counting method and colony formation test， and the cell cycle was measured by flow cytometry. The abilities of cell invasion and migration were examined by Transwell assay. The results showed that SPARC was in perinuclear region and the cytoplasm. Compared with the untransfected cells， cell growth curve and cell colony formation showed that the proliferative ability of SKOV3-PM4-SPARC cells was significantly inhibited (P<0.05). There were no significant differences in cell cycle distribution. The invasion and migration capabilities of SKOV3-PM4-SPARC were also significantly decreased (P<0.05). The abilities of proliferation and invasion of epithelial ovarian cancer cells with directional high lymphatic metastasis were inhibited by SPARC gene over-expression. SPARC gene may play an important role in biological function as a tumor suppressor gene.
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