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二甲双胍通过下调长链非编码 RNA (lncRNA) AFAP1-AS1 和分泌型磷蛋白 1 (SPP1) ，同时上调 miR-3163 来抑制肺腺癌。

Metformin suppresses lung adenocarcinoma by downregulating long non-coding RNA (lncRNA) AFAP1-AS1 and secreted phosphoprotein 1 (SPP1) while upregulating miR-3163.

机构信息

Department of Physical Examination Center, Wuhan Third Hospital, Wuhan, Hubei, China.

Department of Respiratory and Critical Care Medicine, Wuhan Third Hospital, Wuhan, Hubei, China.

出版信息

Bioengineered. 2022 May;13(5):11987-12002. doi: 10.1080/21655979.2021.2005981.

DOI:10.1080/21655979.2021.2005981
PMID:35603556
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9275981/
Abstract

AFAP1-AS1 plays a pro-tumor role in lung cancer. However, no investigation has focused on whether it is involved in the anticancer activity of metformin (Met) in the treatment of lung adenocarcinoma (LUAD). Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to detect the expression of long non-coding (lnc)RNA AFAP1-AS1, the microRNA (miR)-3163, and secreted phosphoprotein 1 (SPP1) in LUAD tissues, or of A549 and H3122 cells. Cell Counting Kit-8, wound scratch, and cell invasion assays were performed to evaluate the effect of the overexpression of lncRNA AFAP1-AS1, miR-3163, and SPP1 on the malignant behaviors of A549 and H3122 cells. Phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) signaling pathway-related proteins were detected by Western blot analysis. Dual luciferase reporter or RIP assays were used to determine the interplay between AFAP1-AS1 and miR-3163, or of miR-3163 and SPP1. Met inhibits the malignant characteristics of A549 and H3122 cells . GEPIA database analysis showed that AFAP1-AS1 is a highly expressed lncRNA in LUAD tissues, which was validated by RT-qPCR. Overexpression of AFAP1-AS1 suppressed the met-mediated anti-tumor activity in A549 and H3122 cells, while AFAP1-AS1 silencing promoted it. Met inhibited AFAP1-AS1 expression, which resulted in reduced proliferation, migration, and invasion in A549 and H3122 cells. This led to AFAP1-AS1-mediated suppression of miR-3163 and, subsequently, the upregulation of SPP1. Met exerts its antitumor activities by regulating the AFAP1-AS1/miR-3163/SPP1/PI3K/Akt/mTOR axis. Our findings deepen our understanding of mechanisms underlying anti-tumor effect of Met in LUAD.

摘要

AFAP1-AS1 在肺癌中发挥促肿瘤作用。然而，尚无研究关注其是否参与二甲双胍（Met）治疗肺腺癌（LUAD）的抗癌活性。采用逆转录定量聚合酶链反应（RT-qPCR）检测 LUAD 组织中长链非编码（lnc）RNA AFAP1-AS1、微小 RNA（miR）-3163 和分泌型磷蛋白 1（SPP1）的表达，或 A549 和 H3122 细胞中的表达。采用细胞计数试剂盒-8（Cell Counting Kit-8，CCK-8）、划痕实验和细胞侵袭实验评估 lncRNA AFAP1-AS1、miR-3163 和 SPP1 过表达对 A549 和 H3122 细胞恶性行为的影响。采用 Western blot 分析检测磷脂酰肌醇 3-激酶/蛋白激酶 B/哺乳动物雷帕霉素靶蛋白（phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin，PI3K/Akt/mTOR）信号通路相关蛋白。采用双荧光素酶报告或 RIP 实验确定 AFAP1-AS1 和 miR-3163 之间的相互作用，或 miR-3163 和 SPP1 之间的相互作用。Met 抑制 A549 和 H3122 细胞的恶性特征。GEPIA 数据库分析显示，AFAP1-AS1 是 LUAD 组织中高表达的 lncRNA，这通过 RT-qPCR 得到验证。AFAP1-AS1 的过表达抑制了 Met 介导的 A549 和 H3122 细胞中的抗肿瘤活性，而 AFAP1-AS1 的沉默则促进了该活性。Met 抑制 AFAP1-AS1 的表达，导致 A549 和 H3122 细胞增殖、迁移和侵袭减少。这导致 AFAP1-AS1 介导的 miR-3163 抑制，随后 SPP1 上调。Met 通过调节 AFAP1-AS1/miR-3163/SPP1/PI3K/Akt/mTOR 轴发挥其抗肿瘤活性。我们的研究结果加深了我们对 Met 在 LUAD 中抗肿瘤作用机制的理解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/5cd43997bea5/KBIE_A_2005981_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/3add181452dc/KBIE_A_2005981_F0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/46d971c303b6/KBIE_A_2005981_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/e03707df5798/KBIE_A_2005981_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/309bfc329eca/KBIE_A_2005981_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/b456f64fd0b9/KBIE_A_2005981_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/cc4ed9bca6ae/KBIE_A_2005981_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/9d31ca87f724/KBIE_A_2005981_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/5cd43997bea5/KBIE_A_2005981_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/3add181452dc/KBIE_A_2005981_F0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/46d971c303b6/KBIE_A_2005981_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/e03707df5798/KBIE_A_2005981_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/309bfc329eca/KBIE_A_2005981_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/b456f64fd0b9/KBIE_A_2005981_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/cc4ed9bca6ae/KBIE_A_2005981_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/9d31ca87f724/KBIE_A_2005981_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f21b/9275981/5cd43997bea5/KBIE_A_2005981_F0008_OC.jpg

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