ADAMDEC1对人脑胶质瘤U87细胞生物学行为的影响

Effects of ADAMDEC1 on the biological behaviour of human glioma U87 cells

目的研究解聚素金属蛋白酶家族癸蛋白1(ADAMDEC1)对人脑胶质瘤U87细胞增殖、黏附、侵袭、迁移和凋亡的影响及可能机制。方法实验组(ADAMDEC1-RNAi)人脑胶质瘤U87细胞用携带特异性siRNA的慢病毒感染来下调ADAMDEC1的表达,对照组(CONTROL-RNAi)用阴性对照慢病毒感染,通过观察GFP荧光的表达情况来判定转染效率,采用Real-time PCR和Western blot法分别从mRNA和蛋白质水平检测稳定转染的U87细胞ADAMDEC1基因的表达情况,CCK-8法检测下调ADAMDEC1表达后细胞增殖及黏附能力的变化,Transwell法检测细胞侵袭及迁移能力的变化,流式细胞技术检测细胞凋亡的变化。结果与CONTROL-RNAi相比,ADAMDEC1-RNAi mRNA及蛋白表达水平均明显降低(P<0.01);CCK-8法检测结果显示下调ADAMDEC1表达能够显著抑制U87细胞的增殖(P<0.05)和黏附能力(P<0.01);Transwell检测结果显示下调ADAMDEC1表达能够显著抑制U87细胞的侵袭和迁移能力(P<0.01);流式细胞术检测结果显示下调ADAMDEC1表达能够促进U87细胞的凋亡(P<0.01)。结论在体外细胞培养试验中,下调ADAMDEC1表达能够显著抑制U87细胞的增殖、黏附、侵袭、迁移能力,促进U87细胞的凋亡。

Objective To investigate the effect of a disintegrin and metalloproteinase like decysin 1 （ADAMDEC1） on proliferation, adhesion, invasion, migration and apoptosis of human glioma U87 cells and its possible mechanism. Methods Human glioma U87 cells in the experimental group （ADAMDEC1-RNAi） were infected with a specific siRNA lentivirns to down regulate the expression of ADAMDEC!, the control group （ CONTROL-RNAi ） was infected with a negative control of lentivirns the transfection efficiency was determined by observing the expression of GFP, Real-time and Western blot methods were used to detect the expression of ADAMDEC1 gene in stably transfected U87 cells from mRNA and protein levels respectively, CCK-8 assay was used to detect the changes of cell proliferation and adhesion after down-regulation of ADAMDEC1 expression, Transwell assay was used to detect the changes of cell invasion and migration, and apoptosis was detected by flow cytometry. Results Compared with the CONTROL-RNAi, the expression level of mRNA and protein in the ADAMDEC1-RNAi were significantly lower （P 〈0. 01 ） ; The results of CCK-8 assay showed that the down-regulation of ADAMDEC1 expression could significantly inhibit the proliferation of U87 cells （P 〈 0.05） and adhesion （P 〈 0. 01 ） ; Transwell detection showed that down-regulated ADAMDEC1 expression could significantly inhibit the invasion and migration of U87 cells （P 〈 0. 01 ） ; The results of flow cytometry showed that down-regulation of ADAMDEC1 expression could promote the apoptosis of U87 cells （P 〈 0. 01 ）. Conclusion In vitro cell culture experiments, down-regulation of ADAMDECl expression can significantly inhibit the proliferation, adhesion, invasion and migration of U87 cells and promote the apoptosis of U87 cells.