Zhu Hui, Wang Shi-Jie, Wang Shun-Xiang, Meng Xian-Li, Wang Yong-Jun
Department of Thoracic Surgery, The Fourth Affiliated Hospital, Hebei Medical University, Shijiazhuang, Hebei 050011, P. R. China.
Ai Zheng. 2005 Dec;24(12):1431-5.
BACKGROUND & 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.
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.
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).
Heparanase gene is expressed in TE-13 cells. Heparanase ASODN can obviously inhibit heparanase gene expression, and restrain invasive activity of TE-13 cells.
乙酰肝素酶是一种内切-D-葡糖醛酸酶，可降解硫酸乙酰肝素蛋白聚糖，在多种恶性肿瘤的侵袭和转移中发挥重要作用。本研究旨在探讨乙酰肝素酶在食管鳞状癌细胞系TE-13中的表达以及乙酰肝素酶反义寡脱氧核苷酸（ASODN）转染对TE-13细胞侵袭的影响。
将合成的乙酰肝素酶ASODN转染至TE-13细胞。转染前后，采用逆转录-聚合酶链反应（RT-PCR）、蛋白质印迹法和免疫细胞化学法检测TE-13细胞中乙酰肝素酶mRNA和蛋白的表达。采用基质胶侵袭试验测定TE-13细胞的侵袭能力。
通过RT-PCR在TE-13细胞中检测到乙酰肝素酶基因（585 bp），通过蛋白质印迹法检测到乙酰肝素酶蛋白（50 ku）。乙酰肝素酶蛋白主要位于细胞质和细胞膜上。乙酰肝素酶ASODN转染后，随着乙酰肝素酶ASODN浓度的升高，乙酰肝素酶基因和蛋白表达以及TE-13细胞的侵袭能力均显著降低（P<0.05，P<0.05）。
乙酰肝素酶基因在TE-13细胞中表达。乙酰肝素酶ASODN可明显抑制乙酰肝素酶基因表达，并抑制TE-13细胞的侵袭活性。
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