Contribution of eIF-4E inhibition to heparanase expression at both mRNA and protein levels in human colon adenocarcinoma cell line LS-174T |
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| Authors: | YANG Yu-jie ZHONG Shi-shun JIAN Yan-ting WAN Tian-mo FU Si-wu LI Xu ZHANG Ya-li |
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| Institution: | Chinese PLA Institute of Digestive Disease, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China |
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| Abstract: | AIM: To determine whether the eukaryotic initiation factor-4E (eIF-4E) inhibition facilitates the degradation of heparanase mRNA and alters heparanase protein expression in human colon adenocarcinoma cell line, LS-174T. METHODS: A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA were introduced into LS-174T cells by lipid-mediated DNA-transfection. eIF-4E protein and mRNA levels were detected by Western blot and RT-PCR, respectively. The mRNA levels of heparanase were determined by Northern blot. The alterations of heparanase expression were confirmed by Western blot analysis. RESULTS: The 20-mer asODN against eIF-4E specifically and significantly inhibited eIF-4E protein expression. Following eIF-4E inhibition, a significant reduction of heparanase mRNA was observed on Northern blot, and at the same time, heparanase protein expression significantly decreased as well. CONCLUSIONS: The results indicate that the inhibition of eIF-4E strongly reduces the stability of heparanase mRNA in colon adenocarcinoma cell line, LS-174T and resultes in an apparent reduction in the expression of heparanase protein. |
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| Keywords: | Colonic neoplasms eIF-4E Heparin lyase LS-174T cells |
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