摘要
目的探讨抑制结肠癌转移相关因子1(MACC1)基因表达对子宫内膜癌细胞增殖凋亡及PI3K/AKT信号通路的影响。方法人子宫内膜癌细胞株Ishikawa被分为空白对照组、阴性对照组和MACC1转染组(转染合成的MACC1-siRNA),转染48 h后,采用Western blot法检测各组细胞MACC1的蛋白表达;CCK8法检测转染24、48、72 h时细胞的增殖情况;流式细胞仪检测转染48 h细胞的凋亡情况;Western blot法检测增殖细胞核抗原(PCNA)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved caspase 3)、磷脂酰肌醇3-激酶(PI3K)、磷酸化的丝氨酸/苏氨酸激酶(p-AKT)的蛋白表达情况。结果阴性对照组与空白对照组的MACC1蛋白相对表达量比较,差异无统计学意义(P>0.05);MACC1转染组的MACC1蛋白相对表达量低于空白对照组,差异有统计学意义(P<0.05)。转染24 h时3组细胞OD_(490)值比较,差异无统计学意义(P>0.05);转染48 h和72 h时,MACC1转染组细胞OD_(490)值均低于空白对照组,差异均有统计学意义(P<0.05)。MACC1转染组细胞凋亡率高于空白对照组,差异有统计学意义(P<0.05)。MACC1转染组PCNA、PI3K、p-AKT蛋白相对表达量均低于空白对照组,cleaved caspase 3蛋白相对表达量高于空白对照组,差异均有统计学意义(P<0.05)。结论抑制MACC1基因表达可降低子宫内膜癌细胞增殖,诱导细胞凋亡,上调cleaved caspase 3蛋白表达,下调PCNA及PI3K/AKT信号通路蛋白的表达。
Objective To investigate the effect of inhibiting metastasis associated in colon cancer 1 （MACC1） gene expression on the proliferation, apoptosis and on PI3K/AKT signaling pathway in endometrial carcinoma.Method Human endometrial carcinoma cell line Ishikawa were stratified as blank control group, negative control group and MACC 1 transfection group （transfected with synthetic MACCI-siRNA）, and then in 48 h after transfection, the protein expression of MACC1 in each group was detected by Western blot; the cell proliferation in 24 h, 48 h, and 72 h after transfection was detected by CCK8 assay; and apoptosis in 48 h after transfection was observed using flow cytometry; the expression of proliferating cell nuclear antigen （PCNA）, cleaved cysteinyl aspartate specific proteinase 3 （cleaved caspase 3）, phosphati- dylinositol 3-kinase （PI3K） and phosphorylated serine / threonine kinase （p-AKT） protein were detected by Western blot.Result The relative protein expression of MACC1 in negative control group and blank control group showed no significant difference （P〉0.05）, while that was significantly lower in MACC1 transfection group compared with blank control group （P〈0.05）. In 24 h after transfection, the OD4~0 values in the three groups were on the same level （P〉0.05）, and after 48 h and 72 h, the value in MACC1 transfection group was significantly reduced compared with the blank control group, the difference was statistically significant （P〈0.05）. As for cell apoptosis, higher apoptosis rate in MACC1 transfection group was observed compared with blank control group, and the difference was statistically significant （P〈0.05）. The relative protein expression of PCNA, PI3K, and p-AKT protein were decreased significantly in MACC1 transfection group compared with blank control group, while the expression of cleaved caspase 3 protein was significantly higher in MACC1 group than in control group, the difference was statistically significant （P〈0.05）.Conclusion Inhib
作者
周斌
孙博
吕会娟
艾亮
古雅丽
ZHOU Bin;SUN Bo;LYU Huijuan;AI Liang;GU Yali(Department of Obstetrics;Clinical Laboratory;Department of Pathology,Zhengzhou Maternal and Child Healthcare Center,Zhengzhou 450000,He＇nan,China)
出处
《癌症进展》
2018年第12期1494-1497,共4页
Oncology Progress
