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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 Kruppel-like factor 4 (KLF4) gene silencing in vitro on proliferation and migration of esophageal carcinoma KYSE140 cells. Methods: The expression of KLF4 protein in human esophageal cancer cell lines KYSE140, KYSE150, EC109 and EC9706 and esophageal immortalized cells NE3 was detected by Western blotting. Two siRNAs targeting KLF4 (KLF4-siRNA1, KLF4-siRNA2) were chemically synthesized The control siRNA (Ctrl-siRNA) was transfected into KLF4-overexpressing KYSE140 cells to form KLF4-siRNA1-KYSE140, KLF4-siRNA2-KYSE140 and Ctrl-siRNA-KYSE140 cells respectively. MTT assay and Transwell assay The proliferation and migration of KYSE140 cells were detected after transfection. Results: The expression of KLF4 protein in esophageal cancer cell line KYSE140 was significantly higher than that in KYSE150, EC109 and EC9706 cell lines (5.62 ± 0.02 vs 1.71 ± 0.23, 3.24 ± 0.35, 3.16 ± 0.41, P < 0.05]. KLF4-siRNA1-KYSE140, KLF4-siRNA2-KYSE140 decreased significantly compared with Ctrl-siRNA-KYSE140 cells (0.49 ± 0.18 vs 0.98 ± 0.19 vs 0.98 ± 0.19, all P <0.05) (P <0.05). The number of migrating cells also increased significantly [(780 ± 22), (475 ± 25) vs (83 ± 17), P <0.05]. Conclusion: KLF4 plays a negative regulatory role in the proliferation and migration of human esophageal cancer cells.