Case Control Study
Copyright ©The Author(s) 2022. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jan 21, 2022; 28(3): 365-380
Published online Jan 21, 2022. doi: 10.3748/wjg.v28.i3.365
Microbiome changes in the gastric mucosa and gastric juice in different histological stages of Helicobacter pylori-negative gastric cancers
Qing-Hua Sun, Jing Zhang, Yan-Yan Shi, Jing Zhang, Wei-Wei Fu, Shi-Gang Ding
Qing-Hua Sun, Jing Zhang, Jing Zhang, Wei-Wei Fu, Shi-Gang Ding, Department of Gastroenterology, Peking University Third Hospital, Beijing 100191, China
Yan-Yan Shi, Research Center of Clinical Epidemiology, Peking University Third Hospital, Beijing 10019, China
Author contributions: Ding SG, Shi YY, and Sun QH designed the study; Sun QH analyzed the data and wrote the manuscript; Sun QH, Zhang J, Fu WW and Zhang J collected the samples and conducted the laboratory experiments; all authors approved the final version of the article.
Supported by National Natural Science Foundation of China, No. 81700496, and No. 81870386; Peking University Medicine Fund of Fostering Young Scholars’ Scientific & Technological Innovation, No. BMU2021PY002; and Key Laboratory for Helicobacter pylori Infection and Upper Gastrointestinal Diseases, Beijing Key Laboratory, No. BZ0371.
Institutional review board statement: The studies involving human participants were reviewed and approved by the Peking University Third Hospital Medical Ethics Committee (No. IRB00006761-M2017414).
Conflict-of-interest statement: No benefits in any form have been received or will be received from a commercial party related directly or indirectly to the subject of this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE Guidelines, and the manuscript was prepared and revised according to the ARRIVE Guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Shi-Gang Ding, MD, Professor, Department of Gastroenterology, Peking University Third Hospital, No. 49 Huayuan North Road, Beijing 100191, China.
Received: November 26, 2021
Peer-review started: November 26, 2021
First decision: December 12, 2021
Revised: December 14, 2021
Accepted: January 11, 2022
Article in press: January 11, 2022
Published online: January 21, 2022

The gastric microbiota in patients with gastric cancer (GC) has received increasing attention, but the profiling of the gastric microbiome through the histological stages of gastric tumorigenesis remains poorly understood, especially for patients with Helicobacter pylori-negative GC (HPNGC).


To characterize microbial profiles of gastric mucosa and juice for HPNGC carcinogenesis and identify distinct taxa in precancerous lesions.


The 16S rRNA gene analysis was performed on gastric mucosa from 134 Helicobacter pylori-negative cases, including 56 superficial gastritis (SG), 9 atrophic gastritis (AG), 27 intestinal metaplasia (IM), 29 dysplasia (Dys), and 13 GC cases, to investigate differences in gastric microbial diversity and composition across the disease stages. In addition, paired gastric mucosa and juice samples from 18 SG, 18 IM, and 18 Dys samples were analyzed. α-Diversity was measured by Shannon and Chao1 indexes, and β-diversity was calculated using partial least squares discrimination analysis (PLS-DA). Differences in the microbial composition across disease stages in different sample types were assessed using the linear discriminant analysis effect size.


The diversity and composition of the bacterial microbiota in the gastric mucosa changed progressively across stages of gastric carcinogenesis. The diversity of the gastric mucosa microbiota was found to be significantly lower in the IM and Dys groups than in the SG group, and the patients with GC had the lowest bacterial community richness (P < 0.05). Patients with IM and those with Dys had similar gastric mucosa microbiota profiles with Ralstonia and Rhodococcus as the predominant genera. Microbial network analysis showed that there was increasing correlation strength between IM and Dys (|correlation threshold|≥ 0.5, P < 0.05). GC and its precancerous lesions have distinguishable bacterial taxa; our results identified HPNGC-associated bacteria Streptococcaceae and Lactobacillaceae (P < 0.05). Additionally, across precancerous lesion stages from AG to Dys in Helicobacter pylori-negative patients, Burkholderiaceae abundance continuously increased, while Streptococcaceae and Prevotellaceae abundance presented a continuous downward trend. Furthermore, the microbial diversity was higher in gastric juice (P < 0.001) than in the mucosa, while PLS-DA revealed a statistically significant difference between the two groups (ANOSIM, P = 0.001). A significant difference in the microbial structure was identified, with Proteobacteria being more prevalent in the gastric mucosa and Firmicutes being more abundant in gastric juice.


Our results provide insights into potential taxonomic biomarkers for HPNGC and its precancerous stages and assist in predicting the prognosis of IM and Dys based on the mucosal microbiota profile.

Keywords: Gastric mucosa, Gastric juice, Microbiota, Stomach neoplasms, Histological stages, 16s RNA gene sequencing

Core Tip: The gastric microbiome profile of Helicobacter pylori-negative precancerous lesions is poorly understood. This is the first study, to our knowledge, to compare the microbiota differences between paired gastric mucosa and gastric juice at different stages of gastric neoplastic progression. The findings revealed that the bacterial community of gastric juice differed from that of the gastric mucosa and that Helicobacter pylori-negative gastric cancer and precancerous lesions have distinct bacterial taxa. Patients with intestinal metaplasia and dysplasia had similar gastric mucosa microbiota profiles, with Ralstonia and Rhodococcus being the most predominant genera, which could aid in prognosis prediction.