Effect of eIF-4E inhibition on heparanase mRNA and its expression in colon adenocarcinoma cell.
OBJECTIVE
To verify whether inhibition of the overexpressed eukaryotic initiation factor-4E (eIF-4E) in human colon adenocarcinoma cell line LS-174T may facilitate the degradation of heparanase mRNA and alter the translation and expression levels of heparanase protein.


METHODS
A 20-mer antisense s-oligodeoxynucleotide (asODN) targeted against the translation start site of eIF-4E mRNA was introduced into LS-174T cells by means of lipid-mediated DNA-transfection, followed by Western blotting analysis and reverse transcription-PCR to determine eIF-4E protein and mRNA levels, respectively. Northern methods was applied to determine heparanase mRNA expression level, with the alterations of heparanase expression assessed by Western blotting analysis.


RESULTS
The 20-mer asODN against eIF-4E specifically and significantly inhibited eIF-4E protein expression, and as a result, a significant reduction in heparanase mRNA level was observed by Northern blotting in conjunction with significantly decreased heparanase protein expression.


CONCLUSIONS
The inhibition of eIF-4E strongly reduces the stability of heparanase mRNA in colon adenocarcinoma cell line LS-174T and results in an apparent reduction in the expression of heparanase protein.
Yang
Yu-jie
YJ
Institute of Digestive Diseases of PLA, Nanfang Hospital, First Military Medical University, Guangzhou 510515, China. yujieyang@fimmu.edu.cn
Zhang
Ya-li
YL
Lai
Zhuo-sheng
ZS
Cui
Hai-hong
HH
Zhong
Shi-shun
SS
Liu
Yu-hu
YH
D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology; D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000201:enzymology* / Q000473:pathology; D039561:Eukaryotic Initiation Factor-4E / Q000037:antagonists & inhibitors*; D005966:Glucuronidase / Q000235:genetics*; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis*; D000099175:Heparanase
D039561:Eukaryotic Initiation Factor-4E; D005966:Glucuronidase; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000099175:Heparanase