Upper gastrointestinal tract microbiota with oral origin in relation to oesophageal squamous cell carcinoma

Figures & data

Table 1. Studies of oral health, tooth loss and periodontitis on esophageal squamous cell carcinoma.

References	Oral health status	Study design	Participant details	Relative risk (95% CI)	Adjustment
Abnet et al. [30]	Tooth loss (Median splits)	Cohort	28,868 person-cohort with 5.25 years follow-up, Linxian, China and 620 incident cases of ESCC	RR:1.3 (1.1–1.6)	Age, sex, tobacco use and alcohol use.
Wang et al. [35]	Tooth brushing (No vs. Yes)	Case–control	116 cases and 189 controls, Linfen, China	OR:0.2 (0.1–0.5)	Age, sex, residence and occupation.
Chen et al. [29]	Tooth loss (6 teeth lost vs. none)	Case–control	616 cases and 770 controls, Taixing, China	OR:1.48 (1.04–2.11)	Age, sex, education, marital status, tobacco use, alcohol consumption, family history, consumption of pickled vegetables and fresh fruits and wealth score.
	Tooth brushing (less than once vs. twice per day)			OR:1.81 (1.37–2.38)
Hiraki et al. [37]	Tooth loss (≥ 21 remaining teeth vs. 0)	Case–control	5,240 cases and 10,480 controls, Aichi Cancer Centre, Japan.	OR:2.36 (1.17–4.75)	Age, sex, smoking and drinking status, vegetable and fruit intake, body mass index and regular exercise.
Dar et al. [31]	Teeth cleaning (≥ once vs. never per day)	Case–control	703 cases and 1,664 controls, Kashmir, India.	OR:0.44 (0.25–0.77)	Age, sex, tobacco and alcohol use, and wealth score.
Abnet et al. [32]	Decayed, missing or filled teeth (32 vs. ≤ 15)	Case–control	283 cases and 560 controls, Golestan, Iran.	OR:2.10 (1.19–3.70)	Age, sex, residence, tobacco and/or opium consumption, alcohol drinking, vegetable intake, socioeconomic status and ethnicity.
	Tooth brushing (Never vs. daily)			OR:2.37 (1.42–3.97)
Sato et al. [34]	Tooth brushing (≥ twice vs. once per day)	Case–control	856 cases of upper aerodigestive tract cancer including 387 oesophageal cases and 2,696 controls, Aichi Cancer Centre, Japan.	OR:0.82 (0.68–0.99)	Age, sex, smoking and drinking status, vegetable and fruit intake, frequent intake of hot beverages, BMI, occupation and number of remaining teeth.
Sepehr et al. [40]	Good oral health vs. dental prosthesis	Case–control	124 cases with oesophageal dysplasia and 50 controls, Turkoman plain, Iran.	OR:4.76 (1.48–15.31)	Age, sex, ethnic origin.
Guha et al. [33]	Tooth loss (≤5 vs. 6-15)	Case–control	91 cases and 566 controls, central Europe.	OR: .84 (1.26–6.41)	Age, sex, education, country, tobacco pack-years, cumulative alcohol consumption.
			95 cases and 359 controls, Latin American	OR:2.18 (1.04–4.59)	Age, sex, education, centre, tobacco pack-years, cumulative alcohol consumption.
Abnet et al. [39]	Tooth loss (0 − 10 vs. 11-31)	Cohort	29,124 person-cohort with 14–year follow-up and 49 incident cases of ESCC.	HR:0.92 (0.46–1.83)	Age, education and H. pylori.
Nwizu et al. [14]	Self-reported periodontal disease	Cohort	65,869 women cohort with 8.32-year follow-up and 34 incident cases of oesophageal cancer	HR:3.28 (1.64–6.53)	Age, smoking status, BMI, history of diabetes and HT use.
Wen et al. [48]	Periodontitis vs. gingivitis	Cohort (Retrospective)	15,3566 participants with 13-year follow-up and 82 incident cases of oesophageal cancer.	HR:1.50 (0.97–2.31)	Age, sex and comorbidities including diabetes, hypertension, hyperlipidaemia.
Chou et al. [49]	Periodontitis (Mild vs. severe)	Cohort	25.485 individuals with mild chronic periodontitis and 25,485 individuals with severe chronic periodontitis using 1:1 propensity score matching.	HR:1.15 (0.62–2.15)	Age, sex, diabetes, cholecystectomy, Charlson comorbidity index score, medication use (aspirin and non-steroidal anti-inflammatory drugs, estimated monthly income and education level.
Michaud et al. [16]	Periodontitis (No vs. yes)	Cohort	48,375 men with median of 17.7-year follow-up and 131 incident cases of oesophageal cancer.	HR:1.44 (0.98–2.11)	Ethnic origin, BMI, physical activity, smoking history, history of diabetes, geographical region, height, alcohol, vitamin D score, calcium intake, fruit and vegetable intake, red-meat intake, and total calorific intake.
	Tooth loss (25-32 vs. 0-16 teeth remaining)			HR:1.34 (0.78-2.30)

Figure 1. Forest Plot for the associations between oesophageal squamous cell carcinoma risk and tooth loss, and poor oral hygiene. The diamonds indicate best estimate of the true (pooled) risk with width indicating 95% confidence intervals (CI). for details, see text.

Table 2. Studies of oral microbiota on oesophageal squamous cell carcinoma.

References	Study design	Participants	Sample type	Method	Results
Chen et al. [69]	Case–control	87 cases with ESCC, 63 individuals with dysplasia and 85 healthy controls from Taixing of Jiangsu, China	Saliva	(V3 to V4 regions of) 16S rRNA gene sequencing	The most common phyla in saliva included Bacteroidetes, Firmicutes, Proteobacteria, Fusobacteria and Actinobacteria; saliva microbiota in ESCC patients harboured the decreased genera including Lautropia, Bulleidia, Catonella, Corynebacterium, Moryella, Peptococcus and Cardiobacterium and the overabundant genera including Prevotella, Streptococcus and Porphyromonas.
Peters et al. [71]	Nested case–control	81 EAC cases and 160 matched controls	Mouthwash	V4 regions 16S rRNA gene sequencing	Saliva microbiota contained overabundance of Tannerella forsythia, and decreased abundances of the commensal genus Neisseria and the species Streptococcus pneumoniae.
		25 ESCC cases and 50 matched controls			Saliva microbiota contained overabundances of Prevotella nanceiensis, Bergeyella oral taxon 322, Neisseria weaveri and Treponema vincentii, and decreased abundances of Prevotella oral taxon 306 and Aggregatibacter paraphrophilus; P. gingivalis was marginally associated with a higher ESCC risk.
Narikiyo et al. [60]	Case–control	20 healthy volunteers and 58 patients with oesophageal cancer from Tokyo, Japan.	Saliva mix	16S rRNA gene amplification and clone sequencing	Streptococcus (S.) mitis (43%), S. sanguinis (13%), S. parasanguis (10%), S. infantis, S. australis, S. constellatus (7%), S. ctirtatus (3%) and unknown Streptococcus species (10%).

Figure 2. The distribution of previously published bacterial taxa derived from oral cavity and oesophagus that represented by the square and circle, respectively. The red/green indicates a detrimental and preventive bacterial taxa. For details, see text.

Table 3. Studies of oesophageal microbiota on oesophageal squamous cell carcinoma.

References	Participants	Sample type	Method	Results
Pei et al. [73]	Four persons with gastrointestinal symptoms	Four distal oesophageal biopsies with normal oesophageal histology	16S rDNA PCR	Six phyla, 41 genera including 13 common genera among 4 persons, and 95 SLOTU including 14 common SLOTU among 4 persons.
Yang et al. [77]	Twelve normal oesophagus, 12 oesophagitis, 10 Barrett’s oesophagus	34 biopsy samples of distal oesophagus.	16S rRNA gene amplification and clone sequencing	Nine phyla, 70 genera and 166 SLOTU; Type I microbiome dominated by Streptococcus in normal oesophagus; Type II microbiome represented by Gram-negative anaerobes in esophagitis and Barrett’s oesophagus
Yu et al. [78]	333 subjects including 142 with ESD and 191 without ESD from Linxian (Linzhou), Henan, China	Upper digestive tract cell samples collected by inflatable rubber balloon covered with cotton mesh or Cytomesh Oesophageal Cytology Device	Human Oral Microbe Identification Microarray for detection of 272 bacterial species.	An inverse association between microbial richness and ESD, and a positive association between microbial richness and PGI/II ratio.
Yang et al.[79]	18 patients with ESCC and 11 volunteers with physiological normal oesophagus from Guangzhou, China	18 ESCC samples and 11 biopsy samples with normal oesophagus	PCR amplification of V4 region of 16S rDNA and sequencing	ESCC tissues had significantly decreased microbial diversity and high abundances of Bacteroidetes, Fusobacteria and Spirochaetes, which were associated with reduced nitrate reductase and nitrite reductase function
Shao et al. [81]	67 patients with ESCC and from Linzhou, Henan, China	Paired ESCC tissue and non-tumour tissue samples	PCR amplification of V4region of 16S rDNA and sequencing	Microbial composition of ESCC comprised of Firmicutes, Bacteroidetes and Proteobacteria phyla; ESCC contained higher abundance of Fusobacteria phylum including Fusobacterium genus, and less abundance of Firmicutes phylum including Streptococcus genus in comparison with non-tumour tissues; ESCC harboured greater abundances of Fusobacteria and Tenericutes phyla, greater abundances of Mycoplasma and Fusobacterium genera in comparison with GCA tumour tissues.
Narikiyo et al. [60]	58 patients with oesophageal cancer from Tokyo, Japan, 4 from China, 2 from France, 5 from Italy.	Oesophageal cancer tissues with or without paired normal tissues.	16S rRNA gene amplification and clone sequencing	Frequent infection of the oral periodontopathic spirochete Treponema denticola, Streptococcus mitis, Streptococcus anginosus in ESCC from different regions of the world.
Yamamura, et al. [62]	325 patients with oesophageal cancer from Kumamoto, Japan	Formalin-fixed, paraffin-embedded (FFPE) oesophageal cancer tissues including 300 ESCCs, 12 adenocarcinomas and 13 of others	qPCR quantification of Fusobacterium nucleatum	F. nucleatum positivity in tumour is 23% (74/325) and is positively correlated with tumour stage. F. nucleatum-positive patients showed poor prognosis with multivariate HR: 1.98 (1.14-3.37)
Yamamura, et al. [63]	551 cases with ESCC	A training cohort of 207 cases with fresh frozen tissues and a validation cohort of 344 cases with FFPE tissues from Nagoya, Kumamoto, Japan, respectively.	qPCR quantification of Fusobacterium nucleatum	High levels of F. nucleatum in ESCC associate with an invasion depth, poor recurrence-free survival, poor chemotherapeutic response
Gao et al. [61]	100 patients with ESCC	100 ESCCs with paired nontumor tissues, and 30 biopsy samples or normal oesophagus from Henan,	Immunohistochemistry and qPCR of 16S rDNA	Porphyromonas gingivalis positivity was 61% in ESCC, 12% in adjacent tissues, and negative in normal oesophagus. High burden of P. gingivalis in ESCC was
		China		positively correlated with poor differentiation, lymph node metastasis, advanced TNM stage, and poor overall survival.
Gao et al. [90]	Patients with ESCC or esophagitis, and healthy controls.	96 cases with ESCC, 50 cases with esophagitis, and 80 healthy controls.	ELISA to measure the serum IgG and IgA antibodies against P. gingivalis.	Serum levels of IgG and IgA against P. gingivalis were higher in ESCC patients than those in patients with esophagitis and healthy controls. Higher levels of IgG and IgA were associated with poor prognosis.

Note: SLOTU denotes species-level operational taxonomic units.

Table 4. Studies of gastric microbiota on esophageal squamous cell carcinoma.

References	Participants	Sample type	Method	Results
Bik et al. [93]	23 adult subjects comprising 12 H. pylori-positive and 11 H. pylori-negative subjects	9 gastric corpus biopsy samples and 14 gastric antrum biopsy samples	16S rRNA gene amplification and clone sequencing	A total of 128 phylotypes were assigned to five major phyla, i.e. Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes and Fusobacteria; no significant association between gastric microbial composition and H. pylori status, gastric anatomical location, and gastric pH value; gastric microbiota is distinct from oral and oesophageal microbiotas.
Nasrollahzadeh et al. [95]	Case cohort comprising 19 patients with stage I to II ESCC and 18 patients with oesophageal squamous dysplasia (ESD), 17 subjects with esophagitis as disease cohort, and 37 normal oesophagus as healthy control cohort from Golestan, Iran	Snap frozen gastric corpus tissue samples	PCR amplification of V3-V4 region of 16S rDNA and sequencing	Five major core phyla across ESCC, ESD, esophagitis and normal oesophagus; higher abundances of Clostridiales and Erysipelotrichales orders in patients with ESCC or ESD; no microbial composition difference between esophagitis and normal oesophagus.
Shao et al. [81]	36 patients with GCA from Linzhou, Henan, China.	Paired GCA tissue and nontumor tissue samples	PCR amplification of V4 region of 16S rDNA and sequencing	Microbial composition of GCA comprised of Firmicutes, Bacteroidetes and Proteobacteria phyla; GCA contained higher levels of Firmicutes, Bacteroidetes, Fusobacteria, Actinobacteria phyla; higher levels of Prevotella, Streptococcus, Veillonella, Haemophilus and Neisseria genera, and lower levels of Proteobacteria phylum and
				Helicobacter genus in comparison with nontumor tissues; GCA had higher abundance of Proteobacteria phylum and Helicobacter genus in comparison with ESCC.
Yu et al. [96]	80 gastric cardia cancer patients from Taiyuan, Shanxi, China	Paired GCA tissue and nontumor tissue samples	PCR amplification of V3-V4 region of 16S rDNA and sequencing	GCA had higher amounts of Bacteriodetes, Firmicutes, Fusobacteria and Spirochaetes phyla, lower amounts of Proteobacteria phylum, Helicobacter genus and H. pylori compared to non-malignant tissues; either H. pylori status or gastric anatomic sites had no effects on gastric microbiota composition;
Yu et al. [97]	80 gastric cardia cancer patients from Taiyuan, Shanxi, China	Paired GCA tissue and non-tumour tissue samples	PCR amplification of V3-V4 region of 16S rDNA and sequencing	In non-malignant tissue, higher level of H. pylori was positively associated with the family history of upper gastrointestinal cancer and the tumour grade; higher abundance of Lactobacillales in patients without metastasis; higher abundance of Bacteroidetes in patients with lower tumour grade.

Abnet CC, Qiao YL, Mark SD, et al. Prospective study of tooth loss and incident esophageal and gastric cancers in China. Cancer Causes Control. 2001;12(9):847–854. doi: 10.1023/a:1012290009545.

 PubMed Web of Science ®Google Scholar

Wang YP, Han XY, Su W, et al. Esophageal cancer in Shanxi province, people’s republic of China: a case-control study in high and moderate risk areas. Cancer Causes Control. 1992;3(2):107–113. doi: 10.1007/BF00051650.

 PubMed Web of Science ®Google Scholar

Chen X, Yuan Z, Lu M, et al. Poor oral health is associated with an increased risk of esophageal squamous cell carcinoma - a population-based case-control study in China. Int J Cancer. 2017;140(3):626–635. doi: 10.1002/ijc.30484.

 PubMed Web of Science ®Google Scholar

Hiraki A, Matsuo K, Suzuki T, et al. Teeth loss and risk of cancer at 14 common sites in Japanese. Cancer Epidemiol Biomarkers Prev. 2008;17(5):1222–1227. doi: 10.1158/1055-9965.EPI-07-2761.

 PubMed Web of Science ®Google Scholar

Dar NA, Islami F, Bhat GA, et al. Poor oral hygiene and risk of esophageal squamous cell carcinoma in Kashmir. Br J Cancer. 2013;109(5):1367–1372. doi: 10.1038/bjc.2013.437.

 PubMed Web of Science ®Google Scholar

Abnet CC, Kamangar F, Islami F, et al. Tooth loss and lack of regular oral hygiene are associated with higher risk of esophageal squamous cell carcinoma. Cancer Epidemiol Biomarkers Prev. 2008;17(11):3062–3068. doi: 10.1158/1055-9965.EPI-08-0558.

 PubMed Web of Science ®Google Scholar

Sato F, Oze I, Kawakita D, et al. Inverse association between toothbrushing and upper aerodigestive tract cancer risk in a Japanese population. Head Neck. 2011;33(11):1628–1637. doi: 10.1002/hed.21649.

 PubMed Web of Science ®Google Scholar

Sepehr A, Kamangar F, Fahimi S, et al. Poor oral health as a risk factor for esophageal squamous dysplasia in northeastern Iran. Anticancer Res. 2005;25(1b):543–546.

 PubMed Web of Science ®Google Scholar

Guha N, Boffetta P, Wünsch Filho V, et al. Oral health and risk of squamous cell carcinoma of the head and neck and esophagus: results of two multicentric case-control studies. Am J Epidemiol. 2007;166(10):1159–1173. doi: 10.1093/aje/kwm193.

 PubMed Web of Science ®Google Scholar

Abnet CC, Kamangar F, Dawsey SM, et al. Tooth loss is associated with increased risk of gastric non-cardia adenocarcinoma in a cohort of Finnish smokers. Scand J Gastroenterol. 2005;40(6):681–687. doi: 10.1080/00365520510015430.

 PubMed Web of Science ®Google Scholar

Nwizu NN, Marshall JR, Moysich K, et al. Periodontal disease and incident cancer risk among postmenopausal women: results from the women’s health initiative observational cohort. Cancer Epidemiol Biomarkers Prev. 2017;26(8):1255–1265. doi: 10.1158/1055-9965.EPI-17-0212.

 PubMed Web of Science ®Google Scholar

Wen BW, Tsai CS, Lin CL, et al. Cancer risk among gingivitis and periodontitis patients: a nationwide cohort study. QJM. 2014;107(4):283–290. doi: 10.1093/qjmed/hct248.

 PubMed Web of Science ®Google Scholar

Chou SH, Tung YC, Wu LS, et al. Severity of chronic periodontitis and risk of gastrointestinal cancers: a population-based follow-up study from Taiwan. Medicine (Baltimore). 2018;97(27):e11386. doi: 10.1097/MD.0000000000011386.

 PubMed Web of Science ®Google Scholar

Michaud DS, Liu Y, Meyer M, et al. Periodontal disease, tooth loss, and cancer risk in male health professionals: a prospective cohort study. Lancet Oncol. 2008;9(6):550–558. doi: 10.1016/S1470-2045(08)70106-2.

 PubMed Web of Science ®Google Scholar

Chen X, Winckler B, Lu M, et al. Oral microbiota and risk for esophageal squamous cell carcinoma in a high-risk area of China. PLoS One. 2015;10(12):e0143603. doi: 10.1371/journal.pone.0143603.

 PubMed Web of Science ®Google Scholar

Peters BA, Wu J, Pei Z, et al. Oral microbiome composition reflects prospective risk for esophageal cancers. Cancer Res. 2017;77(23):6777–6787. doi: 10.1158/0008-5472.CAN-17-1296.

 PubMed Web of Science ®Google Scholar

Narikiyo M, Tanabe C, Yamada Y, et al. Frequent and preferential infection of Treponema denticola, Streptococcus mitis, and Streptococcus anginosus in esophageal cancers. Cancer Sci. 2004;95(7):569–574. doi: 10.1111/j.1349-7006.2004.tb02488.x.

 PubMed Web of Science ®Google Scholar

Pei Z, Bini EJ, Yang L, et al. Bacterial biota in the human distal esophagus. Proc Natl Acad Sci U S A. 2004;101(12):4250–4255. doi: 10.1073/pnas.0306398101.

 PubMed Web of Science ®Google Scholar

Yang L, Lu X, Nossa CW, et al. Inflammation and intestinal metaplasia of the distal esophagus are associated with alterations in the microbiome. Gastroenterology. 2009;137(2):588–597. doi: 10.1053/j.gastro.2009.04.046.

 PubMed Web of Science ®Google Scholar

Yu G, Gail MH, Shi J, et al. Association between upper digestive tract microbiota and cancer-predisposing states in the esophagus and stomach. Cancer Epidemiol Biomarkers Prev. 2014;23(5):735–741. doi: 10.1158/1055-9965.EPI-13-0855.

 PubMed Web of Science ®Google Scholar

Verma D, Garg PK, Dubey AK. Insights into the human oral microbiome. Arch Microbiol. 2018;200(4):525–540. doi: 10.1007/s00203-018-1505-3.

 PubMed Web of Science ®Google Scholar

Shao D, Vogtmann E, Liu A, et al. Microbial characterization of esophageal squamous cell carcinoma and gastric cardia adenocarcinoma from a high-risk region of China. Cancer. 2019;125(22):3993–4002. doi: 10.1002/cncr.32403.

 PubMed Web of Science ®Google Scholar

Yamamura K, Baba Y, Nakagawa S, et al. Human microbiome Fusobacterium Nucleatum in esophageal cancer tissue is associated with prognosis. Clin Cancer Res. 2016;22(22):5574–5581. doi: 10.1158/1078-0432.CCR-16-1786.

 PubMed Web of Science ®Google Scholar

Gao S, Li S, Ma Z, et al. Presence of Porphyromonas gingivalis in esophagus and its association with the clinicopathological characteristics and survival in patients with esophageal cancer. Infect Agent Cancer. 2016;11(1):3. doi: 10.1186/s13027-016-0049-x.

 PubMedGoogle Scholar

Gao SG, Yang JQ, Ma ZK, et al. Preoperative serum immunoglobulin G and a antibodies to Porphyromonas gingivalis are potential serum biomarkers for the diagnosis and prognosis of esophageal squamous cell carcinoma. BMC Cancer. 2018;18(1):17. doi: 10.1186/s12885-017-3905-1.

 PubMedGoogle Scholar

Bik EM, Eckburg PB, Gill SR, et al. Molecular analysis of the bacterial microbiota in the human stomach. Proc Natl Acad Sci USA. 2006;103(3):732–737. doi: 10.1073/pnas.0506655103.

 PubMed Web of Science ®Google Scholar

Nasrollahzadeh D, Malekzadeh R, Ploner A, et al. Variations of gastric corpus microbiota are associated with early esophageal squamous cell carcinoma and squamous dysplasia. Sci Rep. 2015;5(1):8820. doi: 10.1038/srep08820.

 PubMedGoogle Scholar

Yu G, Torres J, Hu N, et al. Molecular characterization of the human stomach microbiota in gastric cancer patients. Front Cell Infect Microbiol. 2017;7:302. doi: 10.3389/fcimb.2017.00302.

 PubMed Web of Science ®Google Scholar

Delgado S, Cabrera-Rubio R, Mira A, et al. Microbiological survey of the human gastric ecosystem using culturing and pyrosequencing methods. Microb Ecol. 2013;65(3):763–772. doi: 10.1007/s00248-013-0192-5.

 PubMed Web of Science ®Google Scholar

Data availability

All data generated or analysed during this study are included in this published article.