摘 要 目的：探讨人肿瘤转移抑制基因KiSS-1对人骨肉瘤MG63细胞侵袭、迁移能力的影响。方法:构建KiSS-1表达质粒pSNAV2.0-KiSS-1。pSNAV2.0-KiSS-1质粒转染骨肉瘤MG63细胞，经G418筛选稳定表达KiSS-1基因的MG63细胞。应用real-time PCR、Western blotting检测KiSS-1 mRNA和蛋白的表达。Transwell小室法检测MG63细胞的侵袭力，millicell小室、细胞划痕愈合实验检测KiSS-1基因对MG63细胞迁移能力的影响。结果:成功建立pSNAV2.0-KiSS-1质粒并稳定转染MG63细胞（MG63-KiSS-1细胞），MG63-KiSS-1细胞高表达KiSS-1 mRNA和蛋白。转染KiSS-1质粒后的MG63细胞侵袭力显著降低（P<0.05）；millicell法、细胞划痕愈合实验证实转染KiSS-1质粒后的MG63细胞迁移力也明显降低（P<0.05）。结论:KiSS-1基因能显著抑制人骨肉瘤MG63细胞的侵袭和迁移能力，在骨肉瘤的转移中起重要作用。

Abstract Objective:To investigate the effects of tumor metastasis-suppressor gene KiSS-1 on the invasion and migration abilities of osteosarcoma MG63 cells. Methods: KiSS-1 expression plasmid pSNAV2.0-KiSS-1 was constructed and transfected into MG63 cells. MG63 cells stably transfected with pSNAV2.0-KiSS-1 (named MG63-KiSS-1) were selected by G418. KiSS-1 mRNA and protein expression in MG63-KiSS-1 cells was examined by real-time PCR and Western blotting analysis, respectively. The invasion ability of MG63 cells was detected by transwell assay. The effects of KiSS-1 on the invasion and migration abilities of MG63 cells were measured by millicell assay and cell scratch healing assay. Results: MG63 cells stably transfected with pSNAV2.0-KiSS-1 were successfully established. MG63-KiSS-1 cells highly expressed KiSS-1 protein. The invasion ability of MG63 cells was significantly decreased after pSNAV2.0-KiSS-1 transfection (P<0.05). The migration ability of MG63 cells was also significantly inhibited after pSNAV2.0-KiSS-1 transfection as examined by millicell assay and cell scratch healing assay (P<0.05). Conclusion: KiSS-1 gene can significantly inhibit the invasion and migration abilities of osteosarcoma MG63 cells, which may play a key role in metastasis of osteosarcoma.

卫生部科学研究基金-福建卫生教育联合攻关计划（No.WKJ2008-2-042）