过表达miR-7对结肠癌HCT-116细胞生物学行为的影响

马利; 熊兵红; 黄卫; 罗华友; 王昆华

doi:10.3971/j.issn.1000-8578.2019.18.0971

过表达miR-7对结肠癌HCT-116细胞生物学行为的影响

Effect of miR-7 Overexpression on Biological Behavior of Colon Carcinoma HCT-116 Cells

摘要

摘要:
目的探讨上调microRNA-7（miR-7）表达对结肠癌HCT-116细胞生物学行为的影响及可能机制。
方法 HCT-116转染后分成miR-7 mimics组、Scramble（阴性对照）组和空白对照组。实时荧光定量PCR（RT-Fq-PCR）检测转染48 h后各组细胞中miR-7表达水平，CCK-8法检测转染72 h后各组细胞的增殖抑制情况，Transwell实验观察转染24 h后细胞侵袭能力，流式细胞仪行细胞周期和细胞凋亡检测。Western blot检测PI3K（p110）、p-Akt及p-mTOR的蛋白表达。
结果 miR-7 mimics组miR-7表达水平明显上调，与其余两组比较差异有统计学意义（P < 0.05）；与阴性对照组和空白对照组相比，miR-7 mimics组细胞增殖抑制率升高，侵袭能力降低，细胞周期分析提示G₀/G₁期的细胞比例增高，S期细胞明显减少，细胞凋亡率明显增加，且PI3K、p-Akt及p-mTOR的蛋白水平明显降低（P < 0.05）。
结论上调结肠癌HCT-116细胞中miR-7表达能抑制肿瘤细胞的增殖与侵袭，其机制可能与抑制PI3K/AKT信号通路有关。

Abstract:
Objective To explore the effect of up-regulation of microRNA-7 (miR-7) on biological behavior of colon carcinoma HCT-116 cells and its possible mechanism.
Methods Colon carcinoma HCT-116 cells were divided into miR-7 mimics group (HCT-116 cells transfected with miR-7 mimics), Scramble (negative control) group (HCT-116 cells transfected with miR-Scramble) and MOCD group (HCT-116 cells without transfection). RT-Fq-PCR was used to test the expression of miR-7 at 48h after transfection. CCK-8 assay was used to test the proliferation of HCT-116 cells at 48h after transfection. Cell invasion ability was determined by Transwell assay at 48h after transfection. Cell cycle and apoptosis rate were detected by flow cytometry. The protein levels of PI3K(p110), p-Akt and p-mTOR were examined by Western blot.
Results The expression of miR-7 was significantly up-regulated after transfection with miR-7 mimics(P < 0.05). The proliferation and metastasis of HCT-116 cells were extremely decreased after transfection with miR-7 mimics. FCM showed that the rate of G₀/G₁ phase was significantly increased, the number of cells in S phase was significantly decreased, the apoptosis rate was significantly upregulated, and the protein levels of PI3K, p-AKT and p-mTOR were decreased in miR-7 mimics group, compared with the other two groups(P < 0.05).
Conclusion Up-regulation of miR-7 could inhibit the proliferation and metastasis of colon carcinoma HCT-116 cells, and the possible mechanism may be related with inhibiting PI3K/AKT signal pathway.

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