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作 者:邹鹰[1,2] 李君[3] 易红[1] 肖艳华[1] 汤参娥[1] 陈主初[1] 肖志强[1]
机构地区:[1]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008 [2]中南大学湘雅医学院组胚教研室,长沙410013 [3]中南大学湘雅医院急诊科,长沙410008
出 处:《国际病理科学与临床杂志》2009年第3期190-195,共6页Journal of International Pathology and Clinical Medicine
基 金:教育部跨世纪优秀人才培养计划基金(教育部科技函[2002]48);湖南省自然科学基金项目(07JJ5031);湖南省科技厅重点科研项目(06SK2004)~~
摘 要: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 silence nm23-H1 gene expression in nasopharyngeal carcinoma (NPC) cell line 6-10B cells by siRNA, and to explore the effect of nm23-H1 downregulation on the biological behavior of 6-10B cells. Methods The nm23-H1 siRNAs and control siRNA were transiently transfected into 6-10B cells by liposome transfection method, respectively. Western blot was used to detect nm23-H1 expression in the transfected cells. MTF assay and flow cytometry were performed to determine the cell growth and cell cycle distribution in the transfected cells, respectively. Transwell system was performed to detect the migration and invasion of the transfected cells. DNA sequencing was used to detect the S120G mutation of nm23-H1 gene in the 6-10B cells. Results Nm23-H1 siRNA could effectively downregulate the expression of nm23-H1 in 6-10B cells, nm23-H1 downregulation could enhance the cell growth, and increase the number of S phase cells and the ability of the in vitro cell migration and invasion in the 6-10B cell line (P 〈0.05). S120G mutation of nm23-H1 gene was not found. Conclusion Nm23-H1 gene behaves as a metastasis suppressor gene.
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