ART1基因沉默对小鼠结肠癌CT26细胞黏附和运动能力的影响

Effects of ART1 gene silencing on the ability of CT26 cellular matrix adhesion and migration

目的探讨单腺苷二磷酸核糖基化转移酶-1[mono(ADP-ribosyl)transferase-1,ART1]基因沉默对小鼠结肠癌CT26细胞黏附、运动和侵袭能力的影响及其机制。方法以携带ART1-shRNA的慢病毒感染小鼠结肠癌CT26细胞。将细胞分为实验组(感染ART1-shRNA慢病毒的CT26细胞),NC-shRNA对照组[感染阴性对照慢病毒(negative-control-shRNA,NC-shRNA)CT26细胞]及未处理组(未经处理的CT26细胞);RT-PCR检测CT26细胞ART1mRNA表达变化,Western blot检测ART1、RhoA、integrinβ1蛋白表达的变化;用细胞基质黏附、运动和侵袭实验观察ART1-shRNA对CT26细胞基质黏附、运动和侵袭能力的影响。结果获得稳定沉默ART1基因的CT26细胞株;RT-PCR和Western blot结果均显示,实验组ART1mRNA和ART1蛋白表达显著低于对照组(P<0.01);Western blot结果显示实验组RhoA、integrinβ1表达均显著低于对照组(P<0.01);实验组细胞黏附、运动、侵袭抑制率分别为70.72%,52.20%和60.00%。结论单腺苷二磷酸核糖基化转移酶-1(ART1)基因沉默可降低CT26细胞的黏附、运动、侵袭能力,该作用可能与ART1沉默后下调RhoA和integrinβ1活性有关。

Objective To explore the effects of mono （ADP-ribosyl）transferase-1 （ART1）gene silencing on the ability of mouse colon carcinoma CT26 cellular matrix adhension, migration and invasion. Methods Lentivirus expressing A RTI-shRNA infected mouse colon carcinoma CT26 cells. The CT26 cells treated with ARTI-shRNA lentivirus were set as experimental group （ART1- shRNA infected group） ;untreated CT26 cells and the CT26 cells treated with negative eontrol-shRNA lentivirus were set as control groups. The expression of ART1 mRNA was detected by RT-PCR. The expression of ART1, RhoA, integrinβ1 were detected by Western blot. The CT26 cells matrix adhesion,migration and invasion potencies were observed by cell matrix adhesion,migration,invasion analysis. Results The CT26 cells with ART1 gene silencing was obtained. Both RT-PCR and Western blot results showed that the expression of ART1 mRNA and ART1 protein in experimental group was significantly lower than that of in control groups （P〈0.01）. The expression of RhoA and integrinβ1 in experimental group was weaker than that in control groups （P 〈0.01）. The inhibitory rates of CT26 cell matrix adhesion, migration and invasion were 70.72%, 52.20%, 60.00%, respectively. Conclusions The knock-down of ART1 in CT26 cell can inhibit cell matrix adhesion, migration, invasion potencies. It may be related to ART1 gene silencing and down-regulating the activity of RhoA and integrinβ1.