阿帕替尼上调miR-324-5p表达抑制结肠癌细胞的研究

Study of apatinib inhibits the colon cancer cells by up-regulating miR-324-5p expression

目的探讨阿帕替尼对结肠癌细胞生物学行为的影响和机制。方法将结肠癌细胞HCT116分为对照(NC)组,低、中、高剂量组(0.5,1.0和2.0μmol·L^(-1)阿帕替尼),转染1组(2.0μmol·L^(-1)阿帕替尼+anti-miR-NC),转染2组(2.0μmol·L^(-1)阿帕替尼+anti-miR-324-5p)。细胞计数试剂盒-8法检测细胞增殖率;Transwell法检测细胞迁移和侵袭;荧光定量聚合酶链反应法检测miR-324-5p表达。结果NC组,低、中、高剂量组、转染1组、转染2组HCT116细胞增殖率分别为(100.00±2.79)%,(94.29±1.74)%,(76.48±1.65)%,(51.48±1.75)%,(51.58±0.88)%和(93.64±3.90)%;迁移细胞数分别为(206.27±9.30),(163.23±3.84),(133.83±4.26),(104.32±5.72),(107.93±3.14)和(194.51±3.96)个;侵袭细胞数分别为(153.47±4.37),(121.42±3.60),(91.99±2.46),(63.37±1.53),(65.95±2.12)和(146.87±3.95)个;miR-324-5p表达水平分别为(1.00±0.09),(1.64±0.10),(1.97±0.13),(2.36±0.17),(1.00±0.11)和(0.45±0.03)。与NC组比较,低、中、高剂量组细胞增殖率、迁移和侵袭数显著降低,miR-324-5p表达水平显著升高,差异均有统计学意义(均P<0.05)。与转染1组比较,转染2组细胞增殖率、迁移和侵袭数显著升高差异均有统计学意义(均P<0.05)。结论阿帕替尼通过上调miR-324-5p的表达来抑制结肠癌细胞增殖、迁移和侵袭。

Objective To explore the effect and mechanism of apatinib on the biological behavior of colon cancer cells.Methods HCT116 colon cancer cells were divided into control(NC)group,low,medium and high dose groups(0.5,1.0 and 2.0μmol·L^(-1) apatinib),transfection group 1(2.0μmol·L^(-1) apatinib+anti-miR-NC),and transfection group 2(2.0μmol·L^(-1) apatinib+anti-miR-324-5p).Cell counting kit-8 method was used to detect cell proliferation rate;Transwell method was used to detect cell migration and invasion;the expression of miR-324-5p was detected by fluorescence quantitative polymerase chain reaction.Results The proliferation rates of HCT116 cells in NC group,low,medium and high dose groups,transfection group 1 and transfection group 2 were(100.00±2.79)%,(94.29±1.74)%,(76.48±1.65)%,(51.48±1.75)%,(51.58±0.88)%and(93.64±3.90)%,respectively.The number of migrated cells was(206.27±9.30),(163.23±3.84),(133.83±4.26),(104.32±5.72),(107.93±3.14),and(194.51±3.96),respectively.The number of invasive cells was(153.47±4.37),(121.42±3.60),(91.99±2.46),(63.37±1.53),(65.95±2.12),and(146.87±3.95),respectively.The expression levels of miR-324-5p were(1.00±0.09),(1.64±0.10),(1.97±0.13),(2.36±0.17),(1.00±0.11)and(0.45±0.03),respectively.Compared with the NC group,the cell proliferation rate,migration and invasion in the low,medium and high dose groups were significantly decreased,and the expression level of miR-324-5p was significantly increased,with statistically significant differences(all P<0.05).Compared with the transfection group 1,the proliferation rate,migration and invasion of cells in the two groups were significantly increased(all P<0.05).Conclusion Apatinib inhibits colon cancer cell proliferation,migration and invasion by upregulating miR-324-5p expression.