miR-21通过调控FOXM1抑制食管癌细胞的迁移及侵袭能力研究

MiR-21 inhibits the migration and invasion of esophageal cancer cells by regulating FOXM1

目的探讨miR-21通过调控叉头框蛋白M1(FOXM1)抑制食管癌细胞的迁移及侵袭能力的效果。方法食管癌细胞系Eca-10在转染时分为3组,miR-21组转染100 pmol miR-21反义核苷酸,对照组转染100 pmol无义核苷酸,空白组转染等剂量的磷酸盐缓冲液。分别于转染24 h与48 h后采用四唑盐比色法(MTT)检测细增殖,Transwell小室检测细胞迁移和侵袭,膜联蛋白V-异硫氰酸荧光素碘化丙啶(Annexin V-FITC/PI)法检测细胞凋亡,蛋白质印迹法检测FOXM1、蛋白激酶B(Akt)蛋白表达。结果转染24 h与48 h后,miR-21组的细胞增殖指数(24 h:35.96±2.48;48 h:45.98±3.11)、迁移(24 h:13.14±1.48;48 h:14.87±2.11)与侵袭指数(24 h:16.98±1.33;48 h:18.77±2.47)、FOXM1(24 h:0.84±0.13;48 h:0.92±0.23)与Akt蛋白(24 h:1.09±0.10;48 h:0.98±0.14)相对表达水平低于对照组与空白组,差异有统计学意义(P<0.05),细胞凋亡指数(24 h:24.92±3.19;48 h:26.09±2.18)高于对照组与空白组,差异有统计学意义(P<0.05),而对照组与空白组以上指标相比,差异无统计学意义(P>0.05)。结论抑制miR-21表达可通过调控FOXM1的表达从而对食管癌细胞的增殖、迁移及侵袭发挥抑制作用,并可促进细胞凋亡。

Objective To investigate the effects of miR-21 on inhibiting the migration and invasion of esophageal cancer cells by regulating forkhead box protein M1(FOXM1).Methods The esophageal cancer cell line Eca-10 were divided into three groups during transfection.The miR-21 group were transfected with 100 pmol miR-21 antisense nucleotides,the control group were transfected with 100 pmol nonsense nucleotides,and the blank group were transfected with the same dose Phosphate buffer.24 h and 48 h after transfection,tetrazolium(MTT)were used to detect cell proliferation,Transwell chamber were used to detect cell migration and invasion,Annexin V-FITC/PI method were used to detect cell apoptosis,and Western blot method were used to detect FOXM1 and protein kinase B(Akt)protein expression.Results At 24 h and 48 h after transfection,the cell proliferation index(24 h:13.14±1.48;48 h:14.87±2.11),migration(24 h:35.96±2.48;48 h:45.98±3.11)and invasion index(24 h:16.98±1.33;48 h:18.77±2.47),relative expression levels of FOXM1(24 h:0.84±0.13;48 h:0.92±0.23)and Akt protein(24 h:1.09±0.10;48 h:0.98±0.14)in the miR-21 group were lower than those in the control group and blank group,the differencec were statistically significant(P<0.05).And the apoptosis index(24 h:24.92±3.19;48 h:26.09±2.18)were higher than the control group,the differencec were statistically significant(P<0.05).There was no statistically significant difference in the above of indicators beteween the control group and blank group(P>0.05).Conclusion Inhibition of miR-21 expression can inhibit the proliferation,migration and invasion of esophageal cancer cells and promote cell apoptosis by regulating the expression of FOXM1.