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Qin-Song Sheng, 1 Kang-Xin He, 2 Jian-Jiong Li, 3 Zi-Feng Zhong, 1 Fei-Xia Wang, 1 Le-Lin Pan, 1 Jian-Jiang Lin 1 1Department of Colorectal and Anal Surgery, the First Affiliated Hospital of College of Medicine, Zhejiang University, Hangzhou, People’s Republic of China; 2State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, the First Affiliated Hospital of College of Medicine, Zhejiang University, Hangzhou, People’s Republic of China; 3Department of Colorectal and Anal Surgery, the First Affiliated Hospital of College of Medicine, Zhejiang University, Ningbo, People’s Republic of ChinaCorrespondence: Jian-Jiong LiDepartment of Colorectal and Anal Surgery, the First Affiliated Hospital of College of Medicine, Zhejiang University, 79 Qingchun Road, Hangzhou 310003, People’s Republic of ChinaTel +86 571 8723 6131Email linjianjiang@zju.edu.cnBackground: To understand the biological effect of gut microbiome on the progression of colorectal cancer (CRC), we sequenced the V3-V4 region of the 16S rRNA gene to illustrate the overall structure of microbiota in the CRC patients.Methods: In this study, a total of 66 CRC patients were dichotomized into different groups based on the following characteristics: paired tumor and adjacent normal tissues, distal and proximal CRC segments, MMR (-) and MMR (+), different TNM staging and clinic tumor staging.Results: By sequencing and comparing the microbial assemblages, our results indicated that 7 microbe genus (Fusobacterium, Faecalibacterium, Akkermansia, Ruminococcus2, Parabacteroides, Streptococcus, and f_Ruminococcaceae) were significantly different between tumor and adjacent normal tissues; and 5 microbe genus (Bacteroides, Fusobacterium, Faecalibacterium, Parabacteroides, and Ruminococcus2) were significantly different between distal and proximal CRC segments; only 2 microbe genus (f_Enterobacteriaceae and Granulicatella) were significantly different between MMR (-) and MMR (+); but there was no significant microbial difference were detected neither in the TNM staging nor in the clinic tumor staging.Conclusion: All these findings implied a better understanding of the alteration in the gut microbiome, which may offer new insight into diagnosing and therapying for CRC patients.Keywords: colorectal cancer, CRC, gut microbiota, 16S rRNA sequencing, TNM staging, MMR