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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 a fibroblast (whip) hair transporter whose function is related to the transduction of signaling pathways. The study by changing the expression level of IFT57 to observe the proliferation of colon cancer SW480 cells to explore the molecular mechanism of IFT57 gene change SW480 proliferation. First, IFT57 overexpression and interference plasmids were constructed and transfected into SW480 colon cancer cells respectively. Realtime PCR and Western blot were used to detect the expression of Gli1, a major transcriptional factor of Hedgehog signaling pathway, and Cyclin D1. Act and Brd U assay SW480 activity and proliferation ability of the situation. The IFT57 high expression plasmid and three sh RNA plasmids were successfully constructed and the one with the highest interference efficiency was screened out (P <0.05). After IFT57 overexpression plasmid was transfected into SW480 cells, the activity of Hedgehog signaling pathway was enhanced and the cell viability and proliferation were enhanced (P <0.05). When IFT57 was silenced, the activity of Hedgehog signaling pathway decreased and the activity and proliferation ability of SW480 cells decreased (P <0.05). These results suggest that IFT57 expression may affect the activity and proliferation of colon cancer SW480 cell line, and its possible mechanism is through the regulation of Hedgehog signaling pathway to affect SW480 cell proliferation activity.