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Updated: Apr 14, 2026
Induction and Analysis of Epithelial to Mesenchymal Transition
Published on: August 27, 2013
Epithelial mesenchymal transition of non-small-cell lung cancer cells A549 induced by SPHK1
Min Ni1, Xiao-Lei Shi1, Zhi-Gang Qu1
1Third Department of Health Care for Cadres, Fuzhou General Hospital of Nanjing Military Region, Fuzhou 350025, China.
Objective:
To explore the effect and molecular mechanism of SPHK1 in the invasion and metastasis process of non-small-cell lung cancer cells (A549).
Methods:
Recombinant retrovirus was used to mediate the production of A549/vector, A549/SPHK1, A549/scramble, and A549/SPHKl/RNAi that stably expressed or silenced SPHK1. The invasion and migration capacities of A549 cells overexpressing or silencing SPHK1 were determined using Transwell invasion assay and scratch wound repair experiment. The protein and mRNA expression levels of E-cadherin, fibronectin, vimentin in A549/vector, A549/SPHK1, A549/scramble, A549/SPHK1/RNAi were detected with Western blot (WB) and quantitative PCR (QPCR) methods, respectively.
Results:
Transwell invasion assay and scratch wound repair experiments showed that over-expression of SPHK1 obviously enhanced the invasion and migration capacities of A549 cells. WB and QPCR detection results showed that, the expression of E-cadherin (a molecular marker of epithelial cells) and fibronectin, vimentin (molecular markers of mesenchymal cells) in A549 cells was upregulated after overexpression of SPHK1; while SPHK1 silencing significantly reduced the invasion and metastasis capacities of A549 cells, upregulated the expression of molecular marker of epithelial cells, and downregulated the expression of molecular marker of mesenchymal cells.
Conclusions:
SPHK1 promotes epithelial mesenchymal transition of non-small-cell lung cancer cells and affects the invasion and metastasis capacities of these cells.
