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目的探讨肿瘤转移抑制基因Kiss-1抑制肿瘤转移的机制。方法通过基因转染技术,将携带Kiss-1基因的pEGFP-N2-Kiss-1质粒转染入YTMLC-90细胞,同时将不携带Kiss-1基因的空质粒转染组和空白对照组作为对照;Millicell小室法检测三组YTMLC-90细胞的侵袭能力,用real time PCR及Western blot方法检测转染后YTMLC-90细胞中Kiss-1、NF-κB及MMP-9 mRNA及蛋白的表达情况。结果成功将Kiss-1基因转染入YTMLC-90细胞后,pEGFP-N2-Kiss-1质粒转染组的侵袭能力较空质粒转染组和空白对照组明显降低,差异有统计学意义(P<0.05);转染了Kiss-1基因的肺鳞癌细胞组Kiss-1 mRNA及蛋白的表达水平明显高于空质粒转染鳞癌细胞组及空白对照组,而NF-κB和MMP-9 mRNA及蛋白的表达水平明显低于空质粒转染鳞癌细胞组及空白对照组,差异均有统计学意义(P<0.05)。结论 Kiss-1基因对YTMLC-90细胞侵袭能力有抑制作用,Kiss-1基因可能通过抑制NF-κB的表达而降低了MMP-9的表达,阻止其对细胞基底膜的降解,从而阻止肿瘤的转移。
Objective To investigate the mechanism of inhibition of tumor metastasis by Kiss-1, a tumor metastasis suppressor gene. METHODS: The pEGFP-N2-Kiss-1 plasmid carrying Kiss-1 gene was transfected into YTMLC-90 cells by gene transfection technique. Meanwhile, the empty plasmid transfection group carrying no Kiss-1 gene and blank control group were used as controls. Millicell chamber method was used to detect the invasion ability of three groups of YTMLC-90 cells. Real time PCR and Western blot were used to detect the expression of Kiss-1, NF-κB and MMP-9 mRNA and protein in transfected YTMLC-90 cells. Results After successfully transfecting Kiss-1 gene into YTMLC-90 cells, the invasive ability of pEGFP-N2-Kiss-1 plasmid transfection group was significantly lower than that of empty plasmid transfection group and blank control group, and the difference was statistically significant (P<0.05). <0.05);Kiss-1 gene transfected lung squamous cell carcinoma group Kiss-1 mRNA and protein expression levels were significantly higher than the empty plasmid transfected squamous cell carcinoma group and blank control group, and NF-κB and MMP-9 The mRNA and protein expression levels were significantly lower than the empty plasmid transfected squamous cell carcinoma group and the blank control group, and the differences were statistically significant (P<0.05). Conclusion The Kiss-1 gene can inhibit the invasive ability of YTMLC-90 cells. The Kiss-1 gene may reduce the expression of MMP-9 by inhibiting the expression of NF-κB and prevent its degradation to the basement membrane of the cells, thus preventing the tumor Transfer.