Streptococcus pneumoniae promotes migration and invasion of A549 cells in vitro by activating mTORC2/AKT through up-regulation of DDIT4 expression

Xiaojie Song; Baohong Liu; Guanghui Zhao; Xiaoxin Pu; Baoyi Liu; Meiling Ding; Yuwen Xue

doi:10.3389/fmicb.2022.1046226

Streptococcus pneumoniae promotes migration and invasion of A549 cells in vitro by activating mTORC2/AKT through up-regulation of DDIT4 expression

Front Microbiol. 2022 Dec 19:13:1046226. doi: 10.3389/fmicb.2022.1046226. eCollection 2022.

Authors

Xiaojie Song¹, Baohong Liu², Guanghui Zhao³, Xiaoxin Pu¹, Baoyi Liu¹, Meiling Ding⁴, Yuwen Xue⁵

Affiliations

¹ Department of Pulmonary and Critical Care Medicine, Qilu Hospital of Shandong University, Qingdao, China.
² Department of Hospital Infection Management, Qilu Hospital of Shandong University, Qingdao, China.
³ Medical Laboratory Center and Oncology Laboratory, Qilu Hospital of Shandong University, Qingdao, China.
⁴ Department of Infectious Diseases, Qilu Hospital of Shandong University, Qingdao, China.
⁵ Department of Pulmonary and Critical Care Medicine, Qilu Hospital of Shandong University, Jinan, China.

Abstract

Introduction: Dysbiosis of the lower airway flora is associated with lung cancer, of which the relationship between Streptococcus, especially pathogenic Streptococcus pneumoniae (S. pneumoniae), and the progression of lung cancer are unclear.

Methods: Bronchoalveolar lavage fluid (BALF) samples were prospectively collected from patients with pulmonary nodules during diagnostic bronchoscopy, and finally included 70 patients diagnosed with primary lung cancer and 20 patients with benign pulmonary nodules as the disease control group. The differential flora was screened by 16S ribosomal RNA (rRNA) gene amplicon sequencing. An in vitro infection model of lung adenocarcinoma (LUAD) cells exposed to S.pneumoniae was established to observe its effects on cell migration and invasion ability. Exploring the molecular mechanisms downstream of DDIT4 through its loss- and gain-of-function experiments.

Results: 16S rRNA sequencing analysis showed that the abundance of Streptococcus in the lower airway flora of lung cancer patients was significantly increased. After exposure to S. pneumoniae, A549 and H1299 cells significantly enhanced their cell migration and invasion ability. The results of DDIT4 loss- and gain-of-function experiments in A549 cells suggest that up-regulation of DDIT4 activates the mTORC2/Akt signaling pathway, thereby enhancing the migration and invasion of A549 cells while not affecting mTORC1. Immunofluorescence (IF) and fluorescence in situ hybridization (FISH) showed that S. pneumoniae was enriched in LUAD tissues, and DDIT4 expression was significantly higher in cancer tissues than in non-cancerous tissues. The increased expression of DDIT4 was also related to the poor prognosis of patients with LUAD.

Discussion: The data provided by this study show that S. pneumoniae enriched in the lower airway of patients with lung cancer can up-regulate DDIT4 expression and subsequently activate the mTORC2/AKT signal pathway, thereby increasing the migration and invasion abilities of A549 cells. Our study provides a potential new mechanism for targeted therapy of LUAD.

Keywords: AKT; DDIT4; lung cancer; mTORC1/2; migration and invasion.