摘要
目的:探讨体外沉默Kruppel样因子4(Kruppel like factor 4,KLF4)基因的表达对食管癌KYSE140细胞增殖及迁移的影响。方法:Western blotting法检测人食管癌细胞株KYSE140、KYSE150、EC109及EC9706及食管永生化细胞NE3中KLF4蛋白的表达,化学合成2对靶向KLF4的siRNA(KLF4-siRNA1,KLF4-siRNA2),并设对照siRNA(Ctrl-siRNA),分别体外转染至高表达KLF4的食管癌KYSE140细胞中,形成KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140及Ctrl-siRNA-KYSE140细胞,通过MTT实验、Transwell实验分别检测转染后食管癌KYSE140细胞的增殖及迁移。结果:食管癌细胞株KYSE140中KLF4蛋白的表达明显高于KYSE150、EC109及EC9706细胞株[(5.62±0.02)vs(1.71±0.23)、(3.24±0.35)、(3.16±0.41),均P<0.05]。KLF4-siRNA1-KYSE140、KLF4-siRNA2-KYSE140与Ctrl-siRNA-KYSE140细胞相比,KLF4蛋白表达明显降低[(0.49±0.18)、(0.32±0.09)vs(0.98±0.19),均P<0.05],细胞增殖能力明显增高[(1.2±0.8)、(1.4±0.1)vs(0.6±0.1),均P<0.05],迁移细胞数量也明显增加[(780±22)、(475±25)vs(83±17)个,P<0.05]。结论:KLF4在人食管癌细胞的增殖和迁移过程中起着负调控作用。
Objective:To investigate the effect of in vitro silencing Kruppel like factor 4(KLF4) gene expression on the proliferation and migration of esophageal cancer KYSE140 cells.Methods: Western blotting was used to detect the expression of KLF4 protein in the esophageal cancer cell lines,including KYSE140,KYSE150,EC109 and EC9706,and immortalized esophageal NE3 cells.Two pairs of siRNAs targeting KLF4(KLF4-siRNA1,KLF4-siRNA2) and control siRNA(Ctrl-siRNA) were chemically synthesized,and were transfected into KYSE140 cells with a high expression of KLF4 in vitro to form KLF4-siRNA1-KYSE140,KLF4-siRNA2-KYSE140 and Ctrl-siRNA-KYSE140 cells.The proliferation and migration of esophageal cancer KYSE140 cells after transfection were detected by MTT assay and Transwell assay,respectively.Results: The expression of KLF4 in KYSE140 cells was higher than that in KYSE150,EC109 and EC9706 cells([5.62±0.02] vs [1.71±0.23],[3.24±0.35],[3.16±0.41],both P0.05).Compared with that in Ctrl-siRNA-KYSE140 cells,the expression of KLF4 protein was significantly decreased in KLF4-siRNA1-KYSE140 and KLF4-siRNA2-KYSE140 cells([0.49±0.18],[0.32±0.09] vs [0.98±0.19],both P0.05),the capacities of proliferation were significantly increased([1.2±0.8],[1.4±0.1] vs [0.6±0.1],both P0.05),and the numbers of migration were also significantly increased([780±22],[475±25] vs ,P0.05).Conclusion: KLF4 functions as a negative regulator in the proliferation and migration of esophageal cancer cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2013年第3期301-305,共5页
Chinese Journal of Cancer Biotherapy
基金
广东省自然科学基金资助项目(No.10451008901005515
No.10151008901000177)~~