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 cancer cells,inhibit cell proliferation and promoting apoptosis.
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)。
