[Inhibitory effect of silencing heparanase expression with antisense oligodeoxynucleotide on invasive capability of human esophageal squamous cancer cell line TE-13].
OBJECTIVE
Heparanase, a kind of endo-D-glucuronidase, degrades heparin sulfate proteoglycans, and plays important roles in invasion and metastasis of many kinds of malignant tumors. This research was designed to investigate the expression of heparanase in esophageal squamous cancer cell line TE-13 and the effect of heparanase antisense oligodeoxynucleotide (ASODN) transfection on invasion of TE-13 cells.


METHODS
Synthesized heparanase ASODN was transfected into TE-13 cells. Before and after transfection, the expression of heparanase mRNA and protein in TE-13 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and immunocytochemistry. Matrigel invasion assay was used to determine the invasive capability of TE-13 cells.


RESULTS
In TE-13 cells, heparanase gene (585 bp) was detected by RT-PCR, and heparanase protein (50 ku) by Western blot. Heparanase protein mainly located in cytoplasm and on cell membrane. After heparanase ASODN transfection, heparanase gene and protein expression and the invasive TE-13 cells were significantly reduced along with the ascending concentration of heparanase ASODN (P<0.05, P<0.05).


CONCLUSIONS
Heparanase gene is expressed in TE-13 cells. Heparanase ASODN can obviously inhibit heparanase gene expression, and restrain invasive activity of TE-13 cells.
Zhu
Hui
H
Department of Thoracic Surgery, The Fourth Affiliated Hospital, Hebei Medical University, Shijiazhuang, Hebei 050011, P. R. China. zhdzuj@yahoo.com.cn
Wang
Shi-Jie
SJ
Wang
Shun-Xiang
SX
Meng
Xian-Li
XL
Wang
Yong-Jun
YJ
D002294:Carcinoma, Squamous Cell / Q000201:enzymology / Q000473:pathology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D004938:Esophageal Neoplasms / Q000201:enzymology / Q000473:pathology*; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D005966:Glucuronidase / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D009361:Neoplasm Invasiveness; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection; D000099175:Heparanase
D005966:Glucuronidase; D020319:Oligodeoxyribonucleotides, Antisense; D012333:RNA, Messenger; D000099175:Heparanase