Abstract
Objective To study the effect of short hairpin RNA (shRNA) targeting extracellular signal-regulated kinase 2 (ERK2) on proliferation of human esophageal cancer cell line Eca109. Methods The recombinant plasmid (pGeneClipTM-shRNA-ERK2) was constructed and transfected into the human esophageal cancer cell line Eca109 by liposome, and the transfection efficiency was observed by reverse fluorescence microscope. The proliferation ability of esophageal cancer Eca109 cells after transfection was analyzed by methyl thiazolyl tetrazolium (MTF). The expression of ERK2 and inhibitor apoptosis gene Survivin following transfection was detected by Western blotting. Results Sequencing proved that recombinant plasmid was successfully constructed, and the transfection efficiency was about 50%-70% after 72 h following transfection. The Eca109 cell growth at 96 h after transfection was significantly inhibited by 10.45% as compared with negative group and blank group,P ＜0. 05. The protein expression of ERK2 and Survivin was decreased in transfection group as compared with the U0126 group and negative group at 72 h and 96 h ( P ＜ 0. 05 ). Conclusion The recombinant plasmid ( pGeneClipTM-shRNA-ERK2) has been proved not only to be effective for down-regulation of ERK2, but also can affect the expression of Survivin and significantly inhibit the Eca109 cells proliferation. Key words: Esophageal carcinoma; Small interference; Extracellular signal-regulated kinase; Cell proliferation
Published Version
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