Twist基因诱导SW480细胞上皮间质转化并增强其侵袭能力的研究

Twist promotes invasion through epithelial-mesenchymal transition in SW480 cells

目的观察Twist基因促进人结肠癌细胞株SW480上皮间质转化及对细胞侵袭能力的影响。方法构建Twist真核表达质粒并转染至SW480细胞。检测3组SW480细胞转染前后Twist、上皮钙黏附蛋白（E-cadherin）和波形蛋白（vimentin）的mRNA和蛋白表达。Transwell小室检测SW480结肠癌细胞的侵袭能力。结果转染Twist基因的SW480细胞TwistmRNA和蛋白表达较未转染组均明显增加（P〈0．01），而E—cadherinmRNA和蛋白表达明显下降（P〈0．01），vimentin的mRNA和蛋白表达明显增加（P〈0．01）。同时转染组SW480细胞侵袭细胞数（91．67±8．37）明显高于转染空质粒组（67．56±6．97）和未转染质粒组（56．67±8．00），差异有统计学意义（P〈0．01）。结论Twist基因通过促进人结肠癌细胞SW480的上皮间质转变而使其具有更强的侵袭能力。

Objective To explore epith＇elial-mesenchymal transition （EMT） in SW480 cells pro- moted by Twist and its effect on invasion ability. Methods The recombinant plasmid pTracer-Twist was constructed. SW480 cells were transfeeted with liposome-mediated plasmid pTraeer-Twist. The expression levels of Twist, E-cadherin and vimentin mRNA and protein before and after transfection were assayed by using real-time quantitative polymerase chain reaction （PCR） and Western blotting in 3 groups. Transwell chambers were used to analyze the invasion ability. Results The mRNA and protein expression levels of Twist in SW480 cells after transfection were increased （ P 〈 0. 01 ）. Significant up-regulation of vimentin and down-regulation of E-cadherin were also observed following transfection of SW480 （ P 〈 0. 01 ）. The number of invasion cells in Twist-transfected group （91.67 ＋ 8. 37 ） was markedly increased as compared with that in non-transfected group （ 56. 67 ＋ 8.00 ） and empty plasmid-transfected group （ 67.56 ＋ 6. 97, P 〈 0. 01 ）. Conclusion Twist enhances invasion ability of SW480 cells in concert with EMT.