T淋巴瘤侵袭转移诱导因子1反义寡核苷酸转染对胃癌细胞形态及侵袭移行能力的影响

目的观察T淋巴瘤侵袭转移诱导因子1（Tiam 1）反义寡核苷酸（ASODN）转染对胃癌细胞形态及体外侵袭、移行能力的影响。方法采用层黏连蛋白黏附法，由胃癌MKN-45细胞株（M0）中筛选获得高侵袭转移亚株（MH）。以脂质体介导将Tiam 1 ASODN转染至MH细胞中，并应用逆转录聚合酶链反应（RT—PCR）及流式细胞技术分别检测Tiam 1 mRNA和蛋白的表达。采用苏木精-伊红染色、细胞骨架蛋白染色、扫描电镜技术及Boyden小室法分别观察转染与未转染MH细胞在形态学及体外侵袭、移行能力方面的变化。结果与对照组相比，应用0．43μmol／L Tiam 1 ASODN转染可特异性抑制胃癌MH细胞中Tiam 1 mRNA和蛋白的表达（P〈0．01）。Tiam 1 ASODN转染MH细胞较未转染MH细胞的体外侵袭、移行能力显著下降（P〈0．05或P〈0．01），同时转染MH细胞较未转染MH细胞膜表面突起及伪足变稀疏或缩短、细胞骨架结构紊乱程度降低、斑点状肌动蛋白小体减少。结论特异性ASODN转染可有效抑制Tiam 1在胃癌细胞中的表达并削弱其体外侵袭、移行能力，这可能是通过调整胃癌细胞骨架结构重组、降低其变形、游走能力而实现的。

Effects of Tiam 1 antisense oligodeoxynucleotides transfection on the morphology and invasive migration potential of gastric cancer cells

OBJECTIVE: To investigate the effects of T lymphoma invasion and metastasis inducing factor 1 antisense oligodeoxynucleotides (Tiam 1 ASODN) transfection on the morphology and invasive migration potential of gastric cancer cells. METHODS: The higher invasive and migratory subgroup (M(H)) were separated from human gastric cancer cell line MKN-45 (M(0)) by laminin adhesion method in vitro. Tiam 1 ASODN was transfected into M(H) cells with liposome, and the expression of Tiam 1 mRNA and protein was determined by RT-PCR and flowcytometry respectivety. The changes in morphology, the invasive and migratory potential between Tima 1 ASODN transfected M(H) cells and no transfected M(H) cells were observed by HE stain, cytoskeletal protein stain, scanning electronic microscope (SEM) and Boyden chamber test. RESULTS: Compared with the control, the expression of Tiam 1 mRNA and protein in M(H) cells was significantly decreased after transfected with 0.43 micromol/L ASODN(P< 0.01). The invasive and migratory potential of M(H) cells in vitro was also much more decreased than that of no transfected cells (P< 0.05 or P< 0.01). At the same time, transfected M(H) cells had less membrane surface projections, fewer or shortener pseudopodia, less irregular cytoskeletal network and less spotted-like actin bodys than no transfected M(H) cells did. CONCLUSION: Tiam 1 ASODN transfection can effectively suppress the expression of Tiam 1 in gastric cancer cells and impair its invasive and migratory potential in vitro, which may be fulfilled through modulating the reconstruction of cytoskeleton and decreasing the deforming and migratory potential of gastric cancer cells.