慢病毒介导的FLOT1基因表达对胃癌细胞侵袭凋亡的机制研究  

Mechanism of Lentivirus Mediated FLOT1 Gene Expression on Invasion and Apoptosis of Gastric Cancer Cells

作　　者：范友杰[1] 王付钦 陈颢[1] FAN You-jie;WANG Fu-qin;CHEN Hao(Department of General Surgery,Xinxiang Central Hospital,Xinxiang 453000,China)

机构地区：[1]新乡市中心医院普外科

出　　处：《中国药学杂志》2019年第16期1299-1304,共6页Chinese Pharmaceutical Journal

摘　　要：目的探讨抑制FLOT1基因表达对胃癌细胞侵袭、凋亡的影响及机制。方法将设计合成的FLOT1siRNA慢病毒载体(si-FLOT1组)转染人胃癌MKN-45细胞,同时转染阴性对照慢病毒载体(阴性组),并设置空白组。Western blotting检测转染48h后细胞FLOT1、E-cadherin、α-SMA、cleaved caspase 3、Bcl-2和Bax蛋白表达。CCK-8法检测转染24、48、72和96h的细胞活力。Transwell小室、流式细胞术及DCFH-DA法分别检测转染48h的细胞侵袭能力、凋亡率及活性氧(ROS)含量。结果FLOT1siRNA慢病毒载体可明显抑制MKN-45细胞FLOT1表达,与空白组比较差异具有统计学意义(P<0.05)。si-FLOT1组与空白组比较,细胞活力降低,细胞侵袭能力降低,细胞凋亡率升高,E-cadherin、cleaved caspase 3和Bax蛋白表达升高,α-SMA和Bcl-2蛋白表达降低,ROS含量升高,差异均具有统计学意义(P<0.05)。结论下调FLOT1基因表达可通过抑制EMT降低胃癌细胞侵袭能力,通过调控凋亡相关蛋白表达及提高细胞ROS含量促进细胞凋亡。OBJECTIVE To investigate inhibitory effect of FLOT1 gene expression on invasion and apoptosis of gastric cancer cells.METHODS The designed FLOT1 siRNA lentiviral vector ( si-FLOT1 group) was transfected into human gastric cancer MKN- 45 cells,at the same time,the negative control lentivirus vector ( negative group) was transfected,and the blank group was set up. Western blotting was used to detect the expression of FLOT1,E-cadherin,α-SMA,cleaved caspase 3,Bcl-2 and Bax proteins.After cells were transfected for 24,48,72 and 96 h,cell viability was detected by CCK-8 assay.After cells were transfected for 48 h,transwell chamber,flow cytometry and DCFH-DA assay were used to detect the invasiveness,apoptosis rate and ROS content,respectively. RESULTS FLOT1 siRNA lentiviral vector inhibited significantly the expression of FLOT1 in MKN-45 cells,which was significantly different from the blank group ( P < 0.05).Compared with si-FLOT1 group and the blank group,the cell vitality decreased,the invasion ability decreased,the apoptosis rate increased,the expression of E-cadherin,cleaved caspase 3 and Bax protein increased, the expression of α-SMA and Bcl-2 protein decreased,and the content of ROS increased,and the difference was statistically significant ( P < 0.05).CONCLUSION Down regulation of FLOT1 gene expression can reduce the invasiveness of gastric cancer cells by inhibiting EMT,and promote apoptosis by regulating the expression of apoptosis related proteins and increasing the ROS content of cells.

关 键 词：胃癌 FLOT1基因 侵袭 凋亡 上皮细胞间质转化 

分 类 号：R965[医药卫生—药理学]