Objective To summarize recent progress in research on intratumoral and local microbiota associated with gallbladder cancer, with a focus on their potential sources, key microbial taxa, possible mechanisms, clinical implications, and methodological challenges, thereby providing evidence for early identification, risk assessment, and microbiota-based intervention studies of gallbladder cancer. Methods Recent studies on gallbladder cancer, biliary tract tumors, and tumor-associated microbiota were reviewed, and evidence from bile microbiota, gallbladder mucosa-associated microbiota, and microorganisms detected in tumor tissues was comparatively analyzed. Results Available evidence suggests that microbiota-associated alterations in gallbladder cancer may be related to retrograde translocation along the gut-biliary axis, hematogenous or portal venous dissemination, and local biliary ecological imbalance. The major taxa reported include Salmonella Typhi, Fusobacterium, Escherichia, Enterococcus, Streptococcus, and Helicobacter. These microorganisms may contribute to gallbladder carcinogenesis through persistent inflammation, DNA damage, bile acid and lipid metabolic reprogramming, immune microenvironment remodeling, and modulation of therapeutic response. However, biliary microbiota, gallbladder mucosa-associated microbiota, and bona fide intratumoral microbiota are not completely equivalent. Low-biomass contamination, host background interference, heterogeneous study designs, and insufficient causal validation remain major limitations in this field. Conclusions Microbiota-related research in gallbladder cancer shows potential clinical value, but direct gallbladder cancer-specific evidence is still limited. Future studies should strengthen standardized sampling, multi-omics integration, and functional validation to improve reproducibility and translational relevance.

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