The Inhibition Effects Of Kiss-1 To The Nasopharyngeal Carcinoma Transplantatiton Tumor In Nude Mice

Objective:(1)Explore the inhibition effect of Kiss-1 gene in nasopharyngeal carcinoma metastasis.(2)Transfer the Kiss-1 gene into the body through transfection technique,and evaluate its application value in the prevention and cure of nasopharyngeal.Methods :(1)Construct the recombinant lentiviral plasmid which contain Kiss-1 gene and transfect the lower expression of Kiss-1 gene,NPC cell line(SUNE-1-5-8F),filtrate is with puromycin,obtain stable cells of Kiss-1 gene high expression and cultivate,passage,cry preserve and resuscitate the stable cells.(2)Test the Kiss-1 protein expression level before and after the transfection with Western blot,and test the Kiss-1 m RNA expression level before and after the transfection with fluoresceace quantitative reverse transcription polymerase chain reaction.(3)Inoculate the oxter of 40 nude mice with nasopharynged carcinoma cells(SUNE-1-5-8F)transfected with Kiss-1 and nasopharynged carcinoma cells non transfected with Kiss-1 respectively.Divide the 40 nude mice into two groups,each group 20 mice,and observe the condition of tumor growth 2-3 times a week.Put the nude mice to death when they were raised to about two months,and dissect the tumors,the livers,the lungs and other organs,carry out an immunohistochemical check on the transplantation tumor,verify whether Kiss-1 have transferred into nasopharyngeal carcinoma or not at the organization level,carry out a pathological examination on the liver,lungs of nude mice.Compare the differences of transplantation tumor between two groups of nude mice,and whether the differences make a difference.Results :(1)We transferred Kiss-1 gene into the SUNE-1-5-8F successfully and completed the cultivation,batches,frozen storage and recovery of transfected cells.(2)Compared to the empty plasmid transfection group and control group,the expression of Kiss-1 protein and m RNA of transfection group significantly increased.In addition,the result of the immunohisto chemical check on the transfected groups and the non transfected groups shows that the Kiss-1 gene in transfected groups is more than that in non transfected groups.Thus Kiss-1 gene successfully transferred into the SUNE-1-5-8Fis verified from the gene,protein,and organization level.(3)The pathological examination on livers,lungs shows the pulmomary metastasis is less in transfected group(one)than that in non transfected group(eight)(p<0.05)Liver metastasis in both two groups of nude mice was not found.There is no statistical significance of two sets nude mice axillary growing tumor.Conclusion: This experiment has used the stable nasopharyngeal carcinoma nude mouse lung transfer model for the research of nasopharyngeal carcinoma and Kiss-1 gene.Active Kiss-1 can be expressed by gene transfection method,and inhibit metastasis of plant tumor.