【摘要】 目的:观察钠氢交换蛋白1(Na~+/H~+ exchanger 1,NHE-1)基因对食管癌KYSE-70细胞增殖和侵袭能力的影响。方法:采用实时荧光定量-PCR和蛋白质印迹法检测NHE-1mRNA及其蛋白在人食管癌EC-9706、KYSE-450、EC-1和KYSE-70细胞中的表达情况。将化学合成的靶向NHE-1的小干扰RNA(small interference RNA,siRNA)体外转染入KYSE-70细胞,实时荧光定量-PCR和蛋白质印迹法检测转染24、48和72h后KYSE-70细胞中NHE-1mRNA及其蛋白的表达,MTT和Boyden小室法检测KYSE-70细胞的增殖及侵袭能力。结果:NHE-1mRNA及其蛋白在KYSE-70细胞中高表达。与阴性对照(转染阴性对照siRNA)及空白对照(未转染的KYSE-70细胞)组相比,靶向NHE-1的siRNA可抑制KYSE-70细胞中NHE-1 mRNA及其蛋白的表达,差异具有统计学意义(P<0.05);与阴性对照组、空白对照组及NHE-1siRNA转染24h组比较,NHE-1siRNA转染48和72h后的KYSE-70细胞体外增殖能力和侵袭能力明显受到抑制,差异有统计学意义(P<0.05)。结论:干扰NHE-1基因的表达可以抑制食管癌KYSE-70细胞的体外增殖和侵袭能力,NHE-1基因有望成为食管癌基因治疗的重要靶点。更多还原

【Abstract】 Objective:To observe the effects of NHE-1 (Na+/H+ exchanger 1) gene on the proliferative and invasive abilities of esophageal cancer cell line KYSE-70. Methods:The expressions of NHE-1 mRNA and protein in esophageal cancer EC-9706, KYSE-450, EC-1 and KYSE-70 cells were detected by real-time fluorescence quantitative-PCR and Western blotting, respectively. Chemically synthesized siRNA (small interference RNA) targeting NHE-1 was transfected into KYSE-70 cells. The expression levels of NHE-1 mRNA and protein in esophageal cancer KYSE-70 cells 24, 48 and 72 h after transfection with NHE-1 siRNA were detected by real-time fluorescence quantitative-PCR and Western blotting, respectively. The proliferative and invasive abilities of KYSE-70 cells were detected by MTT method and Boyden chamber assay, respectively. Results:NHE-1 mRNA and protein were highly expressed in KYSE-70 cells. As compared with the negative control (transfected with nonsense siRNA) and blank control (KYSE-70 cells without transfection), the expression levels of NHE-1 mRNA and protein in KYSE-70 were significantly decreased after transfection with siRNA targeting NHE -1 (all P < 0.05). As compared with the negative control, blank control and the KYSE-70 cells transfected with NHE-1 siRNA for 24 h, the proliferative and invasive abilities of KYSE-70 were inhibited after transfection with NHE-1 siRNA for 48 and 72 h (all P < 0.05). Conclusion:Silencing the expression of NHE-1 gene can inhibit the proliferative and invasive abilities of esophageal cancer KYSE-70 cells in vitro. NHE-1 gene is expected to become an important target for gene therapy of esophageal cancer.更多还原