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目的:采用RNA干扰技术下调nm23-H1基因在鼻咽癌6-10B细胞中的表达,探讨nm23-H1表达下调对6-10B细胞生物学行为的影响。方法:采用脂质体法将nm23-H1基因siRNA(nm23-H1 siRNA)瞬间转染6-10B细胞,Western印迹检测转染细胞中nm23-H1蛋白的表达水平,然后利用MTT法、流式细胞术和Transwell小室实验分别检测转染6-10B细胞的增殖、细胞周期和体外迁移侵袭等生物学行为的变化;测序检测6-10B细胞nm23-H1基因有无S120G点突变。结果:nm23-H1 siRNA有效地下调nm23-H1基因的表达,nm23-H1 siRNA转染6-10B细胞的增殖能力增强,S期细胞增多,体外迁移和侵袭能力增强(P<0.05)。6-10B细胞nm23-H1基因无S120G点突变。结论:nm23-H1基因具有抑制人鼻咽癌细胞6-10B增殖和体外迁移侵袭的作用。
OBJECTIVE: To investigate the effect of nm23-H1 expression down-regulation on the biological behavior of 6-10B cells by down-regulating the expression of nm23-H1 gene in 6-10B cells. Methods: Transfection of nm23-H1 gene siRNA (nm23-H1 siRNA) into 6-10B cells was performed by lipofectamine 2000. The expression of nm23-H1 protein in transfected cells was detected by Western blotting. Then MTT assay, flow cytometry Transwell assay was used to detect the biological behavior of 6-10B cells, such as proliferation, cell cycle and invasion and migration in vitro. Sequencing was used to detect the point mutation of S120G in nm23-H1 gene of 6-10B cells. Results: The nm23-H1 siRNA effectively down-regulated the expression of nm23-H1 gene. The proliferation of 6-10B cells was enhanced by nm23-H1 siRNA. The proliferation of S phase cells was enhanced and the ability of invasion and migration was enhanced in vitro (P <0.05). There is no S120G point mutation in nm23-H1 gene of 6-10B cells. Conclusion: The nm23-H1 gene can inhibit the proliferation and invasion of human nasopharyngeal carcinoma cell line 6-10B in vitro.