miR-100靶向调节食管癌细胞增殖、迁移的机制

Mechanism of miR-100 targeting regulation of proliferation and migration of esophageal cancer cells

目的研究miR-100靶向调节食管癌细胞增殖、迁移的机制。方法培养食管癌Ec-109细胞并进行分组干预,NC模拟物转染Ec-109细胞为NC模拟物组,miR-100模拟物转染Ec-109细胞为miR-100模拟物组,NC抑制物转染Ec-109细胞为NC抑制物组,miR-100抑制物转染Ec-109细胞为miR-100抑制物组。转染24 h后,采用MTS方法检测细胞增殖活力,划痕实验检测细胞迁移活力,Western blot检测细胞中mTOR通路基因的表达水平。结果miR-100模拟物组的OD490值[(0.46±0.07)vs.(0.72±0.12)]、细胞迁移率[(59.81±9.29)%vs.(82.31±12.77)%]以及细胞中哺乳动物雷帕霉素靶蛋白(mTOR)[(0.42±0.09)vs.(0.89±0.14)]、细胞周期蛋白D1 (CyclinD1)[(0.75±0.12)vs.(1.02±0.16)]、B淋巴细胞瘤-2(Bcl-2)[(0.68±0.09)vs.(0.93±0.17)]、基质金属蛋白酶(MMP)2[(0.40±0.07)vs.(0.99±0.13)]、MMP9[(0.52±0.09)vs.(1.08±0.22)]的表达水平均明显低于NC模拟物组,差异均有统计学意义(P<0.05)。miR-100抑制物组的OD490值[(1.21±0.28)vs.(0.80±0.14)]、细胞迁移率[(92.84±16.23)%vs.(80.12±14.28)%]以及细胞中mTOR[(1.38±0.26)vs.(0.77±0.13)]、CyclinD1 [(1.46±0.33)vs.(1.13±0.20)]、Bcl-2[(1.89±0.26)vs.(1.01±0.18)]、MMP2[(2.01±0.42)vs.(1.22±0.16)]、MMP9[(L91±0.35)vs.(1.18±0.19)]的表达水平均明显高于NC抑制物组,差异均有统计学意义(P<0.05)。结论 miR-100能够抑制食管癌细胞的增殖和迁移,靶向抑制mTOR通路是miR-100发挥上述作用的分子机制。

Objective To study the mechanism of miR-100 targeting regulation of proliferation and migration of esophageal cancer cells.Methods Esophageal cancer Ec-109 cells were cultured and divided into NC mimic group,miR-100 mimic group,NC inhibitor group and miR-100 inhibitor group.NC mimic group was transfected with NC mimic,miR-100 mimic group was transfected with miR-100 mimic,NC inhibitor group was transfected with NC inhibitor,and miR-100 inhibitor group was transfected with miR-100 inhibitor.24 hours after transfection,MTS method was used to detect cell proliferation viability,wound-healing assay was used to detect cell migration viability,and Western blot was used to detect the expression level of mTOR pathway gene in cells.Results The OD490 value [(0.46±0.07)vs.(0.72±0.12)],cell mobility[(59.81±9.29)% vs.(82.31±12.77)%] and the expression levels of mammalian target of rapamycin(mTOR) [(0.42±0.09)vs.(0.89±0.14)],CyclinD1 [(0.75±0.12)vs.(1.02±0.16)],B-cell lymphoma-2(Bcl-2) [(0.68±0.09)vs.(0.93±0.17)],matrix metalloproteinase(MMP) 2 [(0.40±0.07)vs.(0.99±0.13)] and MMP9 [(0.52±0.09) vs.(1.08±0.22)] in miR-100 mimic group were significantly lower than those in NC mimic group(P<0.05).The OD490 value [(1.21±0.28) vs.(0.80±0.14)],cell mobility [(92.84±16.23)% vs.(80.12±14.28)%] and the expression levels of mTOR[(1.38±0.26) vs.(0.77±0.13)],CyclinD1 [(1.46±0.33) vs.(1.13±0.20)],Bcl-2[(1.89±0.26) vs.(1.01±0.18)],MMP2[(2.01±0.42) vs.(1.22±0.16)],MMP9 [(1.91±0.35)vs.(1.18±0.19)] in miR-100 inhibitor group were significantly higher than those in NC inhibitor group(P<0.05).Conclusions miR-100 could inhibit the proliferation and migration of esophageal cancer cells.Targeted inhibition of mTOR pathway might be the molecular mechanism by which miR-100 exerts these effects.