SOX9 通过激活 RAP1 信号通路促进肺腺癌细胞的侵袭和迁移。

SOX9 promotes the invasion and migration of lung adenocarcinoma cells by activating the RAP1 signaling pathway.

机构信息

Department of Respiratory and Critical Care Medicine, Taizhou School of Clinical Medicine, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Nanjing Medical University, Taizhou, 225300, Jiangsu, China.

出版信息

BMC Pulm Med. 2023 Nov 2;23(1):421. doi: 10.1186/s12890-023-02740-w.

DOI:10.1186/s12890-023-02740-w

PMID:37919693

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10623714/

Abstract

OBJECTIVE

SOX9 has been shown to be related to the metastasis of various cancers. Recently, it has been reported that SOX9 plays a regulatory role in lung adenocarcinoma (LUAD) cell metastasis, but the specific mechanism remains to be explored. Therefore, the objective of this study was to observe the effect and mechanism of SOX9 on the invasion and migration of LUAD cells.

METHODS

RT-qPCR was applied to observe the expression of SOX9 and RAP1 in tumor tissues and corresponding normal lung tissues collected from LUAD patients. Co-immunoprecipitation and Pearson correlation to analyze the expression correlation of SOX9 with RAP1. To observe the role of SOX9, the invasion and migration levels of LUAD A549 cells in each group were observed by Transwell invasion assay and Scratch migration assay after knocking down or overexpressing SOX9. Besides, the expression levels of RAP1 pathway-related proteins (RAP1, RAP1GAP and RasGRP33) were observed by RT-qCPR or western blot. Subsequently, RAP1 was overexpressed and SOX9 was knocked down in A549 cells, and then the cell invasion/migration level and RAP1 pathway activity were assessed.

RESULTS

The expression levels of SOX9 and RAP1 in tumor tissues and A549 cells of LUAD patients were significantly increased and positively correlated. Overexpression of SOX9 or RAP1 alone in A549 cells enhanced the invasion and migration ability of cells, as well as up-regulated the expression levels of RAP1, RAP1GAP and RasGRP33. However, knocking down SOX9 decreased cell invasion and migration levels and weakened the activity of RAP1 pathway. Notably, overexpressing RAP1 while knocking down SOX9 significantly activated RAP1 pathway and promoted cell invasion and migration.

CONCLUSION

Overexpression of SOX9 in LUAD can significantly activate the RAP1 signaling pathway and promote cell invasion and migration.

摘要

目的

SOX9 已被证明与多种癌症的转移有关。最近有报道称，SOX9 在肺腺癌（LUAD）细胞转移中起调节作用，但具体机制仍需探讨。因此，本研究旨在观察 SOX9 对 LUAD 细胞侵袭和迁移的影响及其机制。

方法

应用 RT-qPCR 观察 LUAD 患者肿瘤组织及相应正常肺组织中 SOX9 和 RAP1 的表达。通过 co-immunoprecipitation 和 Pearson 相关性分析，分析 SOX9 与 RAP1 的表达相关性。通过转染 SOX9 敲低或过表达质粒，观察 SOX9 对 LUAD A549 细胞侵袭和迁移的作用。此外，通过 RT-qCPR 或 Western blot 观察 RAP1 通路相关蛋白（RAP1、RAP1GAP 和 RasGRP33）的表达水平。随后，在 A549 细胞中过表达 RAP1 并敲低 SOX9，评估细胞侵袭/迁移水平和 RAP1 通路活性。

结果

LUAD 患者肿瘤组织和 A549 细胞中 SOX9 和 RAP1 的表达水平均显著升高，且呈正相关。SOX9 或 RAP1 单独过表达均可增强 A549 细胞的侵袭和迁移能力，并上调 RAP1、RAP1GAP 和 RasGRP33 的表达水平。然而，敲低 SOX9 可降低细胞侵袭和迁移水平，并减弱 RAP1 通路活性。值得注意的是，过表达 RAP1 同时敲低 SOX9 可显著激活 RAP1 通路，促进细胞侵袭和迁移。