摘要目的: 探讨微小RNA-218(miR-218)通过磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)/哺乳动物雷帕霉素(mTOR)信号通路对甲状腺癌细胞增殖、迁移和侵袭的影响。方法: 取对数生长期的8505C细胞,分为:对照组(未处理的8505C细胞)、miR-218 NC组(转染阴性mimics)、miR-218 mimics组(转染miR-218 mimics)、通路激活剂组(100 ng/mL PI3K/AKT/mTOR通路激活剂IGF-1)、miR-218 mimics+通路激活组(miR-218 mimics+100 ng/mL IGF-1),检测8505C细胞中增殖蛋白(ki-67)、迁移与侵袭蛋白[基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)]及PI3K/AKT/mTOR信号通路相关蛋白的表达。结果: 与miR-218 NC组比较,miR-218 mimics组8505C细胞中miR-218表达水平[(1.01±0.17) vs (2.31±0.39)]显著上调(P<0.05),OD450值[(2.21±0.07) vs (0.81±0.02)]、细胞划痕愈合率[(34.08±3.17) vs (11.36±0.21)]%、侵袭数目[(81.23±5.68) vs (42.84±2.25)]个及ki-67[(1.68±0.15) vs (0.73±0.02)]、MMP-2[(0.88±0.07) vs (0.39±0.02)]、MMP-9[(2.68±0.27) vs (0.67±0.02)]、p-PI3K[(2.87±0.24) vs (0.36±0.03)]、p-AKT[(0.93±0.05) vs (0.55±0.01)]、p-mTOR蛋白表达[(1.64±0.16) vs (0.47±0.02)]水平显著下调(P<0.05)。结论: miR-218可能通过抑制PI3K/Akt/mTOR信号通路激活抑制8505C细胞增殖、迁移与侵袭。
Abstract:Objective: To investigate the effects of microRNA-218 (miR-218) on the proliferation,migration and invasion of thyroid carcinoma cells through the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Methods: 8505C cells in logarithmic growth phase were divided into:control group (untreated 8505C cells),miR-218 NC group (transfected with negative mimics),miR-218 mimics group (transfected with miR-218 mimics),pathway activator group (100 ng/mL PI3K/AKT/mTOR pathway activator IGF-1),and miR-218 mimics+pathway activator group (miR-218 mimics+100 ng/mL IGF-1).The expression of proliferation protein (ki-67),migration and invasion proteins [matrix metalloproteinase 2 (MMP-2),matrix metalloproteinase 9 (MMP-9)] and PI3K/AKT/mTOR signaling pathway related proteins in 8505C cells were detected. Results: Compared with the miR-218 NC group,the expression level of miR-218 in 8505C cells in the miR-218 mimics group [(1.01±0.17) vs (2.31±0.39)] was significantly up-regulated (P<0.05),the OD450 value [(2.21±0.07) vs (0.81±0.02)],cell scratch healing rate [(34.08±3.17) vs (11.36±0.21)]%,number of invasion [(81.23±5.68) vs (42.84±2.25)],and the protein expression levels of ki-67 [(1.68±0.15) vs (0.73±0.02)],MMP-2 [(0.88±0.07) vs (0.39±0.02)],MMP-9 [(2.68±0.27) vs (0.67±0.02)],p-PI3K [(2.87±0.24) vs (0.36±0.03)],p-AKT [(0.93±0.05) vs (0.55±0.01)],and p-mTOR [(1.64±0.16) vs (0.47±0.02)] were significantly down-regulated (P<0.05). Conclusion: MiR-218 may inhibit the proliferation,migration and invasion of 8505C cells by inhibiting the activation of PI3K/Akt/mTOR signaling pathway.
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