敲减SATB1抑制结肠癌细胞增殖、迁移、侵袭

Inhibitory effect of knockdown SATB1 on proliferation,migration and invasion of colon cancer cells

目的研究慢病毒介导敲减核基质结合区结合蛋白质(SATB)1对结肠癌细胞增殖、凋亡、迁移和侵袭的影响。方法结肠癌SW480细胞感染阴性对照慢病毒和SATB1 shRNA重组慢病毒设置为SW480-sh-NC、SW480-sh-SATB1组,将不进行慢病毒感染的SW480细胞设置为SW480-WT组。Realtime PCR和Western印迹检测敲减效果,噻唑蓝(MTT)方法检测增殖和黏附变化,平板克隆实验检测克隆能力变化,流式细胞术检测细胞凋亡,划痕愈合实验检测迁徙运动能力,Transwell小室检测细胞侵袭和迁移,Western印迹检测钙黏附蛋白E(E-cadherin)、波形蛋白(Vimentin)和侵袭迁移蛋白基质金属蛋白酶(MMP)-2、MMP-9及凋亡蛋白SW480-sh-SATB1组剪切的含半胱氨酸的天冬氨酸蛋白水解酶(Cleaved Caspase)-3、Cleaved Caspase-9表达变化。结果与SW480-sh-NC组比较,SW480-sh-SATB组细胞中SATB1 mRNA和蛋白水平均降低,细胞增殖和克隆能力均下降,细胞凋亡率和细胞中Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平升高,细胞黏附和迁徙运动能力下降,细胞侵袭迁移和细胞中MMP-2、MMP-9蛋白水平降低,细胞中上皮标志物E-cadherin蛋白表达水平升高,间质标志物Vimentin表达水平降低。结论慢病毒介导SATB1敲减抑制结肠癌细胞增殖迁移和侵袭,并诱导细胞凋亡。

Objective To study the effect of lentivirus mediated SATB1 knockdown on proliferation,apoptosis,migration and invasion of colon cancer cells.Methods The negative control lentiviruses and SATB1 shRNA recombinant lentiviruses infected with colon cancer cell SW480 were set as SW480-sh-NC and SW480-sh-SATB1 groups.SW480 cells without lentivirus infection were set as SW480-WT.Realtime PCR and Western blot were used to detecte knockdown effect.The proliferation and adhesion changes were detected by MTT.The cloning ability was detected by plate cloning test.Apoptosis was detected by flow cytometry.Scratch healing test was used to detect migration ability.Transwell cells were used to detect cell invasion and migration.Western blot was used to detect the expression of EMT marker proteins E-cadherin,Vimentin and invasive migration proteins MMP-2,MMP-9,and apoptotic proteins Cleaved Caspase-3 and Cleaved Caspase-9.Results Compared with SW480 cells infected with negative control lentivirus,the levels of SATB1 gene and protein in the cells infected with SATB1 shRNA recombinant lentivirus were decreased,and the cell proliferation and cloning ability were decreased,the apoptosis rate and the expressions of Cleaved Caspase-3 and Cleaved Caspase-9 protein in cells were increased,cell adhesion and migration ability were decreased,cell invasion and migration and MMP-2 and MMP-9 protein levels in cells were decreased,the expression of E-cadherin protein was increased in cells,the expression of interstitial markers Vimentin was decreased.Conclusions Lentivirus mediated SATB1 knockdown could inhibit proliferation,migration and invasion of colon cancer cells and induce apoptosis.