反义抑制Tiam 1对胃癌细胞形体重塑影响的实验研究

Effects of Tiam 1 Antisense Oligonucleotides on Morphological Remodeling of Gastric Cancer Cells

目的观察T淋巴细胞瘤侵袭转移诱导因子1(Tiam1)反义寡核苷酸(ASODN)转染对胃癌细胞形体重塑的影响。方法采用层粘连蛋白黏附法,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Tiam1ASODN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Tiam1mRNA和蛋白的表达。采用HE染色、细胞骨架蛋白染色及扫描电镜技术观察转染与未转染MH细胞在形态学方面的不同。结果与未转染组相比,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);转染MH细胞的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Tiam1在胃癌细胞中的表达及胃癌细胞形体重塑,这可能对胃癌细胞的侵袭转移产生影响。

Objective To investigate the effects of T lymphoma invasion and metastasis inducing factor 1 （Tiara 1） antisense oligonucleotides （ASODN） on morphological remodeling of gastric cancer cells. Methods The high-invasive and metastastic subgroup （MH） was separated from human gastric cancer cell line MKN-45 （MH） by laminin adhesion method in vitro. And they were divided into four groups according to different further treatments： no transfection group （control group）, liposome transfection group, sense oligonucleotides-liposome transfection group （SODN transfection with liposome group） and antisense oligonucleotides-liposome transfection group （ASODN transfection with liposome group）. Then the expressions of Tiara 1 mRNA and protein were detected by RT-PCR and flowcytometry, respectively. The morphology changes between Tima 1 ASODN transfected MH cells and no transfected cells were observed by using HE stain, cytoskeletal protein stain and scanning electronic microscope （SEM）. Results Compared with the other groups, the expressions of Tiara 1 mRNA and protein in MH cells were significantly decreased after the cells were transfeeted with 0. 43μmol/L Tiara 1 ASODN （P〈0. 01）. Addi- tionally, it was observed that the transfected MH cells had less membrane surface projections, fewer or shortener pseudopodia, less irregular cytoskeletal network and less spotted-like actin bodys than no transfected MH cells did. Conclusion ASODN transfection could effectively suppress the expression of Tiara 1 and the remodeling in gastric cancer cells, which may play an important role in the invasion and metastasis of gastric cancer cells.