Effects and Related Mechanisms of Truncated LEF-1 Isoforms on Biological Behavior of HeLa Cell Lines

Abstract:Objective To study the effects of truncated LEF-1 isoforms on the biological behavior of HeLa cell lines and the related mechanisms. Methods Truncated LEF-1 gene was obtained by PCR from human's lymphoid node cDNA library and inserted into pCDNA3.1/V5-His vector to construct the eukaryotic expression plasmid pcDNA3.1-Δ-LEF-1.Using LipofectamineTM 2000，the plasmid pcDNA3.1-Δ-LEF-1 or empty vector was transfected into Hela cells.Then the stable cell lines which expressed the truncated LEF-1 isoforms was screened by G418 and expression of target protein was identified by Western blot.Subsequently, the proliferation，apoptosis, cell cycle and expression of CXCR4 of transgenic cell lines were analyzed by flow cytometry.In addition, the capability of adhesion and migration of transgenic cell lines were investigated by extracellular matrigel adhesion assay and transwell assay respectively. Results The truncated LEF-1 eukaryotic expression plasmid and the stable HeLa cell lines expressing the truncated LEF1 isoforms were constructed successfully.The proliferation of the transgenic cell lines was inhibited, but their apoptosis was increased, meanwhile the cells were blocked at G0/G1 stage.The ability of adhesion and migration of the transgene cell lines were reduced and the expression of CXCR4 was upregulated. Conclusion The truncated LEF--1 isoforms can inhibit the proliferation and migration of HeLa cells, and promote their apoptosis, which maybe depend on the expression CXCR4 partly.