摘要
目的探讨肌成束蛋白(Fascin)-1介导自噬对子宫内膜癌细胞增殖与凋亡的影响。方法对数生长期的子宫内膜癌HEC-2B细胞分为3组:对照组、实验1组与实验2组,3组分别转染同体积的磷酸盐缓冲液、siRNA空载体、siRNA-Fascin-1载体。采用MTT法检测细胞增殖情况,流式细胞仪检测细胞凋亡情况,免疫印迹法(Western Blotting)检测LC3Ⅱ、Fascin-1蛋白表达水平,定量聚合酶链式反应(qPCR)检测LC3Ⅱ、Fascin-1 mRNA表达水平。结果转染后24 h与36 h,实验2组的细胞增殖指数分别为(45.00±4.19)%、(54.09±5.87)%,显著低于对照组与实验1组,实验2组的细胞凋亡指数分别为(18.92±2.21)%、(20.14±2.83)%,显著低于对照组与实验1组,差异有统计学意义(P<0.05);实验1组与对照组比较差异无统计学意义(P>0.05)。转染后24 h与36 h,实验2组的LC3Ⅱ mRNA与蛋白表达水平分别为3.98±0.12、4.24±0.23;5.76±0.13、5.77±0.20,高于对照组与实验1组,Fascin-1 mRNA与蛋白表达水平分别为1.87±0.13、1.92±0.14;1.22±0.03、1.27±0.12,低于对照组与实验1组,差异有统计学意义(P<0.05),实验1组与对照比较差异无统计学意义(P>0.05)。结论在子宫内膜癌细胞中抑制Fascin-1表达能促进LC3Ⅱ在转录水平与翻译水平的表达,从而提高子宫内膜癌细胞自噬水平,抑制细胞增殖与促进细胞凋亡。
Objective To investigate the effects of fascin-1 mediated autophagy on proliferation and apoptosis of endometrial cancer cells.Methods Endometrial cancer HEC-2B cells in logarithmic growth phase were divided into three groups: the control group,experiment group 1 and experiment group 2.The control group,experiment group 1 and experiment group 2 were transfected with the same volume of phosphate buffer,siRNA empty vector,siRNA-Fascin-1 vector.Cell proliferation were detected by MTT method,apoptosis were detected by Annexin V-FITC&PI,LC3Ⅱ and Fascin-1 protein expression levels were detected by Western Blotting,and LC3Ⅱ and Fascin-1 mRNA expression levels were detected by qPCR.Results At 24 h and 36 h after transfection,the cell proliferation index of the experimental group 2 were(45.00 ± 4.19) %,(54.09 ± 5.87) %,were significantly lower than the control group and the experimental group 1,the differences were statistically significant(P<0.05),and the apoptotic index were(18.92 ± 2.21) %,(20.14 ± 2.83) %,were higher than the control group and the experimental group 1,the differences were statistically significant(P<0.05).There were no significant difference in the compared between the control group and the experimental group 1(P>0.05).Compared with the control group and the experimental group 1,the expression of LC3Ⅱ mRNA and protein in the experimental group 2 were 3.98 ± 0.12,4.24 ± 0.23;5.76 ± 0.13,5.77 ± 0.20,were higher than the control group and the experimental group 1(P<0.05).The expression of Fascin-1 mRNA and protein were 1.87 ± 0.13,1.92 ± 0.14;1.22 ± 0.03,1.27± 0.12,were significantly lower than the control group and the experimental group 1,the differences were statistically significant(P<0.05).There were no significant difference compared between the experimental group 1 and the control group(P>0.05).Conclusion Inhibition of Fascin-1 expression can promote the expression of LC3Ⅱ at the transcriptional level and translation level,thereby increase the level of autophagy in endometrial
作者
高艳飞
韩宏斌
陈强
GAO Yan-fei;HAN Hong-bin;CHEN Qiang(Department of Laboratory Medicine,Panjin Liaoyou Gem Flower Hospital,Panjin Liaoning 124010,China;Department of Oncology,Panjin Liaoyou Gem Flower Hospital,Panjin Liaoning 124010,China)
出处
《临床和实验医学杂志》
2020年第22期2396-2399,共4页
Journal of Clinical and Experimental Medicine
基金
辽宁省科学技术计划项目(编号:2018225040)。
