miR-101-3p在胃癌中的表达及其靶向STC-1基因调控PI3K/AKT信号通路对癌细胞侵袭转移和血管生成的影响

刘强; 孙少明; 王文俊

doi:10.3971/j.issn.1000-8578.2022.22.0609

miR-101-3p在胃癌中的表达及其靶向STC-1基因调控PI3K/AKT信号通路对癌细胞侵袭转移和血管生成的影响

Expression of miR-101-3p in Gastric Cancer and Its Effects on Invasion, Metastasis, and Angiogenesis of Gastric Cancer Cells by Targeting STC-1 Gene to Regulate PI3K/AKT Signaling Pathway

摘要

摘要:
目的探究miR-101-3p在胃癌中的表达及其靶向STC-1基因调控PI3K/AKT信号通路对癌细胞侵袭转移、血管生成的作用机制。
方法 qRT-PCR检测胃癌组织及BGC-823细胞中miR-101-3p和STC-1 mRNA的表达，并分析miR-101-3p表达与患者临床病理因素的关系；通过Lipofectamine^TM 2000将miRNA模拟物和质粒分别或者联合转染细胞；TargetScanHuman预测及双荧光素酶报告验证miR-101-3p对STC-1的靶向调控关系；划痕实验、Transwell小室实验、Matrigel体外成管实验及Western blot检测验证miR-101-3p靶向STC-1基因对癌细胞的侵袭转移和血管生成的影响及可能的机制，并通过裸鼠致瘤实验检测移植瘤的发展。
结果胃癌组织中STC-1的表达水平高于正常组织。与正常胃组织和GES-1细胞相比，胃癌组织和BGC-823细胞中miR-101-3p下调，STC-1 mRNA上调，且miR-101-3p水平与STC-1水平负相关，miR-101-3p水平与肿瘤分化程度、TNM分期、淋巴结转移显著相关（P < 0.05）；过表达miR-101-3p可抑制STC-1的表达，下调p-PI3K/PI3K、p-AKT/AKT、MMP-2、MMP-9、VEGF、Ang2水平，抑制肿瘤细胞的侵袭转移和血管生成，降低移植瘤体积和重量（P < 0.05）。
结论 miR-101-3p在胃癌中表达下调，能够靶向STC-1基因调控PI3K/AKT信号通路抑制胃癌细胞BGC-823的侵袭转移和血管生成及体内移植瘤的进展。

Abstract:
Objective To explore the expression of miR-101-3p in gastric cancer and its mechanism on the invasion, metastasis, and angiogenesis of gastric cancer cells by targeting the STC-1 gene to regulate the PI3K/AKT signaling pathway.
Methods qRT-PCR was used to detect the expression of miR-101-3p and STC-1 mRNA in gastric cancer tissues and BGC-823 cell and analyze the relationship between miR-101-3p expression and patients' clinical pathological factors. The cells were transfected with miRNA mimics and plasmids separately or in combination with Lipofectamine^TM 2000. TargetScanHuman prediction and dual-luciferase assay were used to verify the targeted regulation of miR-101-3p on STC-1. The effect and possible mechanism of miR-101-3p targeting the STC-1 gene on the invasion, metastasis, and angiogenesis of cancer cells were verified by scratch test, Transwell chamber test, Matrigel in vitro tube forming test, and Western blot assay. The development of the transplanted tumor was detected by nude mouse tumorigenicity test.
Results The expression of STC-1 in gastric cancer tissues was higher than that in normal tissues. Compared with normal gastric tissues and GES-1 cells, miR-101-3p was down-regulated, and STC-1 mRNA was up-regulated in gastric cancer tissues and BGC-823 cell. The level of miR-101-3p was negatively correlated with the level of STC-1, and significantly correlated with the degree of tumor differentiation, TNM stage, and lymph node metastasis (P < 0.05). miR-101-3p directly targeted STC-1. The overexpression of miR-101-3p inhibited STC-1 expression and downregulated the expression of p-PI3K/PI3K, p-AKT/AKT, MMP-2, MMP-9, VEGF, and Ang2, consequently, inhibited tumor cell invasion, metastasis, and angiogenesis and reduced the size and weight of the transplanted tumors (P < 0.05).
Conclusion miR-101-3p is down-regulated in gastric cancer and can target the STC-1 gene to regulate the PI3K/AKT signaling pathway and inhibit the invasion, metastasis, and angiogenesis of BGC-823 gastric cancer cells and the development of transplanted tumors in vivo.

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