Effects of S100A6 gene silencing on proliferation and migration of human esophagus cancer cell line Eca109
LU Wen-qua
Abstract
LU Wen-qua
Abstract
Objective To investigate the effects of S100A6gene silencing on the proliferation and migration of Eca109 human esophagus cancer cells.Methods The shRNA expression vectors were constructed using the shRNA sequences designed based on human S100A6's coding sequence,and were transfected into Eca109cells via cationic liposome.The changes of S100A6mRNA and protein in Eca109cells transfected with the recombinant vectors were detected using real-time PCR and Western blotting analysis 48hours after transfection,respectively;the proliferative curves of transfected cells were plotted using MTT assay;furthermore,the change in cellular migration ability was determined using wound healing assay. Results The eukaryotic expression vector of shRNA targeting S100A6 was successfully constructed.Real-time PCR and Western blotting analysis results showed that S100A6 was effectively silenced by liposome-mediated transfection of the recombinant shRNA vectors in Eca109cells.Compared with the untransfected cells,S100A6mRNA and protein in transfected Eca109cells were significantly decreased(P0.05,P0.01).Meanwhile,the proliferative activity of Eca109cells was significantly inhibited by S100A6silencing(P0.01).It was found that the cellular migration was also suppressed by S100A6 gene interference.Conclusion S100A6gene can be effectively silenced by shRNA expression vectors,and the silence may lead to inhibition of the proliferation and migration of Eca109cells.
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Objective To investigate the effects of S100A6gene silencing on the proliferation and migration of Eca109 human esophagus cancer cells.Methods The shRNA expression vectors were constructed using the shRNA sequences designed based on human S100A6's coding sequence,and were transfected into Eca109cells via cationic liposome.The changes of S100A6mRNA and protein in Eca109cells transfected with the recombinant vectors were detected using real-time PCR and Western blotting analysis 48hours after transfection,respectively;the proliferative curves of transfected cells were plotted using MTT assay;furthermore,the change in cellular migration ability was determined using wound healing assay. Results The eukaryotic expression vector of shRNA targeting S100A6 was successfully constructed.Real-time PCR and Western blotting analysis results showed that S100A6 was effectively silenced by liposome-mediated transfection of the recombinant shRNA vectors in Eca109cells.Compared with the untransfected cells,S100A6mRNA and protein in transfected Eca109cells were significantly decreased(P0.05,P0.01).Meanwhile,the proliferative activity of Eca109cells was significantly inhibited by S100A6silencing(P0.01).It was found that the cellular migration was also suppressed by S100A6 gene interference.Conclusion S100A6gene can be effectively silenced by shRNA expression vectors,and the silence may lead to inhibition of the proliferation and migration of Eca109cells.
Key concepts: Transfection, Small hairpin RNA, Gene silencing, Blot, Molecular biology, Cell growth, RNA interference, Cell culture