Abstract
Background LIM and SH3 domain protein 1 (LASP1), highly expressed in a variety of tumors, is considered as a novel tumor metastasis biomarker. However, it is unknown which signaling pathway works and how the signal transduces into cell nucleus to drive tumor progression by LASP1. The aim of this study is to explore the essential role of LASP1 in TGF-β1-induced epithelial-mesenchymal transition (EMT) in lung cancer cells. Methods The gene and protein levels of LASP-1 were successfully silenced or overexpressed by LASP-1 shRNA lentivirus or pcDNA in TGF-β1-treated lung cancer cell lines, respectively. Then, the cells were developed EMT by TGF-β1. The cell abilities of invasion, migration, and proliferation were measured using Transwell invasion assay, wound healing assay, and MTT assay, respectively. Western blotting was used to observe the protein levels of EMT-associated molecules, including N-cadherin, vimentin, and E-cadherin, and the key molecules in the TGF-β1/Smad/Snail signaling pathway, including pSmad2 and Smad2, pSmad3 and Smad3, and Smad7 in cell lysates, as well as Snail1, pSmad2, and pSmad3 in the nucleus. Results TGF-β1 induced higher LASP1 expression. LASP1 silence and overexpression blunted or promoted cell invasion, migration, and proliferation upon TGF-β1 stimulation. LASP1 also regulated the expression of vimentin, N-cadherin, and E-cadherin in TGF-β1-treated cells. Activity of key Smad proteins (pSmad2 and pSmad3) and protein level of Smad7 were markedly regulated through LASP1. Furthermore, LASP1 affected the nuclear localizations of pSmad2, pSmad3, and Snail1. Conclusion This study reveals that LASP1 regulates the TGF-β1/Smad/Snail signaling pathway and EMT markers and features, involving in key signal molecules and their nuclear levels. Therefore, LASP1 might be a drug target in lung cancer.
Highlights
Targeted therapy and new targets provide new hope for the treatment of non-small-cell lung cancer (NSCLC) lung cancer [1]
During coinfection with 2 ng/ml TGF-β1 and LIM and SH3 domain protein 1 (LASP1) pcDNA, LASP1 mRNA (Figures 2(a) and 2(c)) and protein (Figures 2(b) and 2(d)) levels were all significantly increased compared to the TGF-β1 group (p < 0.05). e differences among groups of TGF-β1, SC Small Hairpin RNA (shRNA), and pcDNA were not obvious (p > 0.05). ese results proved that TGFβ1 stimulated the increased expression of LASP1, the lentivirus containing LASP1 shRNA still inhibited the induced LASP1, whereas LASP1 pcDNA enhanced the further expression. e cell models could be used for the experiments
Since the epithelial-mesenchymal transition (EMT)-related downstream molecules can be regulated at the transcriptional level and nuclear translocation occurs in the TGF-β1/ Smad/Snail1 signaling pathway, we investigated the nuclear levels of pSmad2, pSmad3, and Snail1. e lack of α-tubulin and the presence of Lamin B demonstrated the purified nuclear extracts in these preparations (Figure 6(a))
Summary
Targeted therapy and new targets provide new hope for the treatment of NSCLC lung cancer [1]. LIM and SH3 domain protein 1 (LASP1), highly expressed in a variety of tumors, is considered as a novel tumor metastasis biomarker. It is unknown which signaling pathway works and how the signal transduces into cell nucleus to drive tumor progression by LASP1. E aim of this study is to explore the essential role of LASP1 in TGF-β1induced epithelial-mesenchymal transition (EMT) in lung cancer cells. E gene and protein levels of LASP-1 were successfully silenced or overexpressed by LASP-1 shRNA lentivirus or pcDNA in TGF-β1-treated lung cancer cell lines, respectively. Is study reveals that LASP1 regulates the TGF-β1/Smad/Snail signaling pathway and EMT markers and features, involving in key signal molecules and their nuclear levels. Conclusion. is study reveals that LASP1 regulates the TGF-β1/Smad/Snail signaling pathway and EMT markers and features, involving in key signal molecules and their nuclear levels. erefore, LASP1 might be a drug target in lung cancer
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