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目的:从基因—蛋白—细胞效应多层次水平,观察Tiam-1反义基因下调肿瘤细胞内转移相关基因Tiam-1 mRNA和调节细胞骨架结构的Rho蛋白表达水平,探讨遏制肿瘤侵袭转移的作用和机制。方法:采用反义核酸技术合成Tiam-1反义寡核苷酸,硫代化修饰与脂质体形成复合物(Tiam-1 ASODNS-Lf),转导入人高转移巨细胞肺腺癌PG细胞。采用RT-PCR技术检测Tiam-1的表达水平,流式细胞术检测Rho蛋白表达水平,Matrigel体外转移模型检测PG细胞的侵袭转移力。结果:Tiam-1 ASODNS-Lf抑制PG细胞Tiam-1 mRNA表达水平,其相对表达值从0.86下降为0.38;显著抑制PG细胞内Tiam-1介导的信号传递相关Rho蛋白表达水平,其表达率从27.96％降至16.86％(P<0.001)。显著抑制穿过人工基底膜的侵袭转移PC细胞数,从945±40下降为649±35(P<0.05)。结论:Tiam-1反义基因下调PG细胞中侵袭诱导基因mRNA表达及其介导的信号传递蛋白Rho的表达,从而遏制其侵袭和转移能力。
OBJECTIVE: To observe the effect of Tiam-1 antisense gene down-regulating the expression of Tiam-1 mRNA and regulating the cytoskeleton structure of Rho protein in the multi-level of gene-protein-cell effect, and to explore the role of Tiam-1 in inhibiting tumor invasion and metastasis mechanism. Methods: Tiam-1 antisense oligonucleotide was synthesized by antisense oligonucleotide. Tiam-1 ASODNS-Lf was modified by liposome and then transduced into PG cells of human high metastatic giant cell lung adenocarcinoma . The expression of Tiam-1 was detected by RT-PCR. The expression of Rho protein was detected by flow cytometry. The invasion and metastasis of PG cells were detected by Matrigel in vitro. Results: Tiam-1 ASODNS-Lf inhibited the expression of Tiam-1 mRNA in PG cells, and the relative expression of Tiam-1 ASODNS-Lf decreased from 0.86 to 0.38. Tiam-1-mediated signal transduction-related Rho protein expression in PG cells was significantly inhibited From 27.96% to 16.86% (P <0.001). Significantly inhibited the invasion and metastasis of PC cells through the artificial basement membrane, from 945 ± 40 to 649 ± 35 (P <0.05). Conclusion: Tiam-1 antisense gene down-regulates the expression of invasion-induced gene mRNA and the expression of Rho in PG cells, thereby inhibiting its invasion and metastasis.