改变IFT57表达水平对结肠癌SW480细胞增殖的影响及其分子机制

The Effect of IFT57 Expression Level Change on the Proliferation of Colon Carcinoma SW480 Cells and Its Molecular Mechanism

IFT57是一种纤(鞭)毛内转运蛋白,其功能与信号通路的转导相关。该研究通过改变IFT57表达水平后,观察其对结肠癌SW480细胞增殖能力的影响,探讨IFT57基因改变SW480增殖能力的分子机制。首先,构建IFT57过表达及干扰质粒,分别转染结肠癌SW480细胞株后,采用Realtime PCR及Western blot技术检测Hedgehog信号通路关键转录因子Gli1、主要靶基因Cyclin D1的表达水平变化;采用CCK-8法及Brd U法检测SW480活性及增殖能力的情况。成功构建了IFT57高表达质粒及3个sh RNA质粒并筛选出干扰效率最高的1个(P<0.05)。将IFT57过表达质粒转染SW480细胞株后,Hedgehog信号通路活性升高,细胞活性和增殖能力增强(P<0.05);当沉默IFT57表达后,Hedgehog信号通路活性下降,细胞的活性及增殖能力降低(P<0.05)。以上结果提示,IFT57表达水平可影响结肠癌SW480细胞株活性及增殖能力,其可能机制是通过调控Hedgehog信号通路活性来影响SW480细胞的增殖活力。

IFT57 is one of intraflagellar transport protein in cilia, whose function is implicated with the conduction of signaling pathway. The aim of this study was to investigate the effect on viability and proliferation of colon carcinoma SW480 and explore the possible molecular mechanism when the expression level of IFT57 was altered in SW480 cells. We altered the expression level of IFT57 in SW480 cells by transfecting pc DNA3.1-IFT57 or sh RNA pc DNA6.2-IFT57. The key molecular Gli1 and target gene Cyclin D1 of Hedgehog signaling pathway were detected by Real-time PCR and Western blot. The cell viability and proliferation of SW480 were determined by CCK-8 and Brd U assay, respectively. The results showed that the activity of Hedgehog signaling pathway had been changed along with the expression level of IFT57（P〈0.05）. The viability and proliferation of SW480 were increased along with the elevated expression level of IFT57（P〈0.05）, and SW480 was inhibited after the decreasedexpression level of IFT57（P〈0.05）. Our results indicated that IFT57 could alter the viability and proliferation of SW480, and the possible molecular mechanism might alter the activity of Hedgehog signaling pathway to contribute the viability and proliferation ability of SW480 cells.