沉默ADAM17对HT29人结肠癌细胞体外侵袭能力的影响及机制探究

Effect of silencing ADAM17 on invasion ability of HT29 human colon cancer cells

摘要:

目的 探究沉默解聚素-金属蛋白酶17（ADAM17）对HT29人结肠癌细胞体外侵袭能力的影响及机制。方法 HT29人结肠癌细胞经慢病毒转染处理后沉默ADAM17蛋白的表达，实时聚合酶链反应（real-time PCR）检测ADAM17 mRNA的表达，单层细胞划痕损伤修复实验检测细胞迁移能力的变化，Transwell体外侵袭实验检测细胞侵袭能力的改变，western blotting检测转染后MMP-9蛋白及ERK表达情况。结果 经慢病毒转染处理后HT29细胞的ADAM17表达被抑制（P?<0.05）。与空白对照组比较，沉默ADAM17的表达后，转染组细胞的迁移和侵袭能力被抑制（P?<0.05）；沉默ADAM17后，细胞基质金属蛋白酶9（MMP-9）表达水平下调、磷酸化细胞外调节蛋白激酶（p-ERK1/2）表达水平下降（P?<0.05），但细胞外调节蛋白激酶（ERK）表达水平差异无统计学意义（P?>0.05）。结论 ADAM17可能通过抑制ERK通路激活下调MMP-9的表达，从而影响细胞迁移、侵袭能力。

Abstract:

Objective To observe the effects of ADAM17 on the invasion of human colon cancer cell lines HT29. Methods The expression of ADAM17 protein was silenced by slow virus transfection. Silencing of ADAM17 was confirmed by real-time PCR and Western blot. Cell migration was measured by wound healing assay. The expression of MMP-9 and ERK was detected by Western blotting. Results The expression of ADAM17 protein in HT29 cells was significantly inhibited through silencing technology (P??0.05). Conclusions ADAM17 may promote cell migration and invasion by inhibiting ERK pathway mediated expression of MMP-9.