Bacterial proteomic profile of early colonizers on the acquired pellicle of patients with head and neck cancer

VENTURA, Talita Mendes Oliveira; PEREIRA, Júlia Thomazini; OLIVEIRA, Ramilli Maria de; THOMASSIAN, Larissa Tercília Grizzo; RIBEIRO, Nathalia Regina; TAIRA, Even Akemi; PELÁ, Vinícius Taioqui; VALLE, Aline Dionizio; RUBIRA, Cássia Maria Fisher; SANTOS, Paulo Sérgio da Silva; BUZALAF, Marília Afonso Rabelo

doi:10.1590/1807-3107bor-2026.vol40.055

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Sumário

Original Research/Cariology • Braz. oral. res. 40 • 2026 • https://doi.org/10.1590/1807-3107bor-2026.vol40.055 linkcopy

Bacterial proteomic profile of early colonizers on the acquired pellicle of patients with head and neck cancer

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Radiotherapy is the cornerstone of head and neck cancer (HNC) treatment, but it has been frequently associated with changes in the acquired enamel pellicle (AEP), potentially affecting the early stages of dental biofilm formation. Thus, this study aimed to characterize the bacterial proteomic profiles of early colonizers adhering to the AEP in patients with HNC undergoing radiotherapy, with emphasis on radiation-induced changes that may influence oral biofilm development and oral health outcomes. AEP samples were collected from nine patients with HNC before radiotherapy (BRT), during radiotherapy (DRT), and after radiotherapy (ART), as well as from nine orally healthy individuals. Pellicles were obtained 120 min after formation and analyzed by shotgun label-free proteomics using nanoLC-ESI-MS/MS. Protein expression levels were compared between groups using the t test (p < 0.05). AEP showed elevated levels of bacterial proteins predominantly attributed to Actinomyces, mainly associated with DNA binding, chromatin organization, and glucose metabolism. The bacterial proteomic profile shifted markedly DRT, with exclusive detection of proteins involved in carbohydrate transport and metabolism, oxidative stress response, and protein folding, including several ATP-binding cassette transporters and glycolytic enzymes. Proteins related to stress tolerance, transcriptional regulation, and secretion systems were detected ART, many of them associated with bacterial species commonly linked to periodontal diseases, such as Tannerella and Oribacterium. Notably, Cysteine synthase was identified exclusively BRT in all comparisons, suggesting its potential as a biomarker. These findings indicate that radiotherapy induces dynamic changes in the AEP bacterial proteome, potentially favoring dysbiotic biofilm development and increasing susceptibility to oral diseases in patients with HNC.

Descriptors
Head and Neck Neoplasms; Radiotherapy; Dental pellicle; Biofilms; Proteomics

Access number	Protein name*	PLGS score	Ratio	p-values	Protein function**
Access number	Protein name*	PLGS score	BRT:Control	p-values	Protein function**
L1PFP6	Glyceraldehyde-3-phosphate dehydrogenase OS=Actinomyces sp. oral taxon 181 str. F0379	455	3.56	< 0.01	Glucose metabolic process;
					glyceraldehyde-3-phosphate dehydrogenase (NAD+) activity (phosphorylation);
					NAD binding; NADP binding
E7NDS1	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 171 str. F0337	696	3.13	< 0.01	DNA binding;
E7NDS1		696	3.13	< 0.01	structural component of chromatin
F3PBV0	DNA-binding protein HU-alpha OS=Actinomyces sp. oral taxon 170 str. F0386	696	3.13	0.02	DNA binding; structural component of chromatin
G9PMW1	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 849 str. F0330	696	3.06	< 0.01	DNA binding, structural component of chromatin
F9EHV8	DNA-binding protein HU OS=Actinomyces sp. oral taxon 448 str. F0400	696	3.00	< 0.01	DNA binding; structural component of chromatin
A0A2S0LKX0	Integration host factor OS=Actinomyces sp. oral taxon 897	696	2.94	< 0.01	DNA binding; structural component of chromatin; chromosomal condensation
F9PMQ7	Cysteine synthase OS=Actinomyces sp. oral taxon 175 str. F0384	BRT*	-	-	Cysteine synthase activity;
F9PMQ7		BRT*	-	-	cysteine biosynthesis from serine
D3IAR7	dTDP-4-dehydrorhamnose 3_5-epimerase OS=Prevotella sp. oral taxon 299 str. F0039	Control*	-	-	Dtdp-4-dehydroramnose 3,5-epimerase; dTDP-rhamnose biosynthetic process
I8UL97	Methyltransferase domain protein OS=Capnocytophaga sp. oral taxon 412 str. F0487	Control*	-	-	Methyltransferase activity dependent on S-adenosylmethionine; methylation
A0A2S0L8X1	Methyltransferase OS=Capnocytophaga sp. oral taxon 878	Control*	-	-	Methyltransferase activity dependent on S-adenosylmethionine; methylation
D3IGK9	Replication initiation protein OS=Prevotella sp. oral taxon 317 str. F0108	Control*	-	-	Replication initiation proteins are necessary to open the double DNA helix and initiate DNA replication in bacteria
A0A2S0L935	tRNA-specific 2-thiouridylase MnmA OS=Capnocytophaga sp. oral taxon 878	Control*	-	-	Catalytic activity mediating the unidirectional 2-thiolation of the positive swing (U34) of tRNA, leading to the formation of s2U34
L1PPF2	Uncharacterized protein OS=Capnocytophaga sp. oral taxon 324 str. F0483	Control*	-	-	Uncharacterized protein
L1PDJ0	Uncharacterized protein OS=Capnocytophaga sp. oral taxon 324 str. F0483	Control*	-	-	Activity of methyltransferase dependent on S-adenosylmethionine
L1Q0W6	Uncharacterized protein OS=Capnocytophaga sp. oral taxon 326 str. F0382	Control*	-	-	Activity of methyltransferase dependent on S-adenosylmethionine
S3BMC1	Uncharacterized protein OS=Capnocytophaga sp. oral taxon 336 str. F0502	Control*	-	-	Uncharacterized protein
S3BHQ6	Uncharacterized protein OS=Capnocytophaga sp. oral taxon 336 str. F0502	Control*	-	-	Activity of methyltransferase dependent on S-adenosylmethionine
A0A0K1F0W8	Uncharacterized protein OS=Olsenella sp. oral taxon 807	Control*	-	-	Uncharacterized protein
A0A2T4TJP5	Uncharacterized protein OS=Prevotella sp. oral taxon 313	Control*	-	-	Uncharacterized protein

Access number	Protein name	PLGS score	Unique*	p-value	Function
F9PMQ7	Cysteine synthase OS=Actinomyces sp. oral taxon 175 str. F0384	91	BRT	NC	Cysteine synthase activity, cysteine biosynthesis from serine
D0WNJ5	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 848 str. F0332	987	BRT	NC	DNA binding, structural constituent of chromatin
G9PMW1	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 849 str. F0330	987	BRT	NC	DNA binding, structural constituent of chromatin
F9EHV8	DNA-binding protein HU OS=Actinomyces sp. oral taxon 448 str. F0400	987	BRT	NC	DNA binding, structural constituent of chromatin
F3PBV0	DNA-binding protein HU-alpha OS=Actinomyces sp. oral taxon 170 str. F0386	987	BRT	NC	DNA binding, structural constituent of chromatin
A0A0M4GFM1	60 kDa chaperonin OS=Actinomyces sp. oral taxon 414	127	DRT	NC	Capsule, cell surface, cytoplasm, ATP binding, ATP-dependent protein folding companion, isomerase activity, unfolded protein binding, protein doubling
A0A2T4T1K6	3-deoxy-D-manno-octulosonate 8-phosphate phosphatase OS=Prevotella sp. oral taxon 820	137	DRT	NC	Hydrolase activity, acting on ester bonds, metal ion binding
G9PJA1		80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity and carbohydrate transport
G9PJA1	ABC transporter domain-containing protein OS=Actinomyces sp. oral taxon 849 str. F0330	80	DRT	NC
G5F356	ABC transporter domain-containing protein OS=Olsenella sp. oral taxon 809 str. F0356	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
U2EIT0	ABC transporter_ ATP-binding protein OS=Clostridiales bacterium oral taxon 876 str. F0540	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
H1LYP5	ABC transporter_ ATP-binding protein OS=Lachnospiraceae bacterium oral taxon 082 str. F0431	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
F5T447	ABC transporter_ ATP-binding protein OS=Oribacterium sp. oral taxon 108 str. F0425	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
E0Q2F5	ABC transporter_ ATP-binding protein OS=Streptococcus sp. oral taxon 071 str. 73H25AP	80	DRT	NC	ATP binding cassette carrier complex (ABC), ABC type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
L1PB56	Conjugative transposon protein TraI OS=Capnocytophaga sp. oral taxon 324 str. F0483	69	DRT	NC	Conjugative Transposon Protein TraI
A0A0M4H7M4	Fructose-1_6-bisphosphate aldolase OS=Actinomyces sp. oral taxon 414	199	DRT	NC	Fructose-biphosphate aldolase activity, glycolytic process
F9EDG0	Fructose-bisphosphate aldolase OS=Actinomyces sp. oral taxon 448 str. F0400	200	DRT	NC	Fructose-biphosphate aldolase activity, glycolytic process
A0A0M4GT11	L-lactate dehydrogenase OS=Actinomyces sp. oral taxon 414	179	DRT	NC	Cytoplasm, L-lactate dehydrogenase activity, glycolytic process
A0A2S0LU24	Lipocalin OS=Dietzia sp. oral taxon 368	121	DRT	NC	Proteins that bind to small hydrophobic molecules, such as lipids and retinoids. They transport lipids, vitamins, hormones, and metabolites. Involved in immune responses, cellular migration and differentiation. Associated with the interactions of cancer cells and biological synthesis of prostaglandins.
F3B1Q7	Maltodextrin import ATP-binding protein msmX OS=Lachnospiraceae oral taxon 107 str. F0167	80	DRT	NC	ATP binding cassette carrier complex (ABC), ABC type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
S2ZT92	Maltose/maltodextrin transport system ATP-binding protein OS=Atopobium sp. oral taxon 199 str. F0494	80	DRT	NC	ATP binding cassette carrier complex (ABC), ABC type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
A0A1B3X4Z9	Mannitol-1-phosphate 5-dehydrogenase OS=Selenomonas sp. oral taxon 920	112	DRT	NC	Mannitol-1-phosphate 5-dehydrogenase activity, mannitol metabolic process
L1PRA4	Nitroreductase family protein OS=Capnocytophaga sp. oral taxon 324 str. F0483	80	DRT	NC	Oxidoreductase activity
A0A2R3N0E1	Sugar ABC transporter ATP-binding protein OS=Lachnospiraceae bacterium oral taxon 500	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
A0A0K1F3A3	Sugar ABC transporter ATP-binding protein OS=Olsenella sp. oral taxon 807	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
A0A0X8JXI3	Sugar ABC transporter ATP-binding protein OS=Streptococcus sp. oral taxon 431	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
F9EDZ0	Sugar ABC superfamily ATP binding cassette transporter_ ABC protein OS=Actinomyces sp. oral taxon 448 str. F0400	111	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
A0A0M4GDJ0	Sugar ABC transporter ATP-binding protein OS=Actinomyces sp. oral taxon 414	111	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
F9ED26	Thioredoxin reductase OS=Actinomyces sp. oral taxon 448 str. F0400	163	DRT	NC	Cytoplasm, thioredoxine-disulfide reductase (NADP) activity, removal of superoxide radicals
F9HGK8	TOBE domain protein OS=Streptococcus sp. oral taxon 056 str. F0418	80	DRT	NC	ATP binding carrier complex (ABC), ABC-type transporter activity, ATP binding, ATP hydrolysis activity, and carbohydrate transport
A0A2N8NJX9	Uncharacterized protein OS=Tannerella sp. oral taxon 808	40	DRT	NC	Uncharacterized protein
W2CLQ4	Uncharacterized protein OS=Tannerella sp. oral taxon BU063 isolate Cell 6/7/9	56	DRT	NC	Uncharacterized protein
D3IJU8	Uncharacterized protein OS=Prevotella sp. oral taxon 317 str. F0108	80	DRT	NC	Uncharacterized protein
U2M781	Uncharacterized protein OS=Selenomonas sp. oral taxon 892 str. F0426	139	DRT	NC	Uncharacterized protein

Access number	Protein name	PLGS score	*Unique	p-value	^bFunction
F9PMQ7	Cysteine synthase OS=Actinomyces sp. oral taxon 175 str. F0384	91	BRT	NC	Cysteine synthase activity, cysteine biosynthesis from serine
F9PJ25	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 175 str. F0384	987	BRT	NC	Histone-like DNA-binding protein capable of wrapping around DNA to stabilize it and thus prevent its denaturation under extreme environmental conditions
F9EHV8	DNA-binding protein HU OS=Actinomyces sp. oral taxon 448 str. F0400	987	BRT	NC	DNA binding, structural constituent of chromatin
F3PBV0	DNA-binding protein HU-alpha OS=Actinomyces sp. oral taxon 170 str. F0386	987	BRT	NC	DNA binding, structural constituent of chromatin
G9PMW1	DNA-binding protein HB1 OS=Actinomyces sp. oral taxon 849 str. F0330	987	BRT	NC	DNA binding, structural constituent of chromatin
A0A0M4H4B4	Integration host factor OS=Actinomyces sp. oral taxon 414	987	BRT	NC	DNA binding, structural constituent of chromatin
U2AR85	DUF1963 domain-containing protein OS=Capnocytophaga sp. oral taxon 863 str. F0517	116	ART	NC	DUF1963 domain-containing proteins that are basic units that can fold, function, and evolve independently
U2W4E1	Late embryogeneis abundant protein OS=Oribacterium sp. oral taxon 078 str. F0263	111	ART	NC	They protect cells from damage caused by environmental stresses, such as changes in temperature, salinity, and dryness, by providing buffer hydration.
W2CIT0	Secretion system C-terminal sorting domain-containing protein OS=Tannerella sp. oral taxon BU063 isolate Cell 1/3	114	ART	NC	C-terminal sorting domain-containing protein of the secretion system
W2C4L3	Secretion system C-terminal sorting domain-containing protein OS=Tannerella sp. oral taxon BU063 isolate Cell 2	114	ART	NC	C-terminal sorting domain-containing protein of the secretion system
W2CA78	Secretion system C-terminal sorting domain-containing protein OS=Tannerella sp. oral taxon BU063 isolate Cell 5	114	ART	NC	C-terminal sorting domain-containing protein of the secretion system
W2CMU8	Secretion system C-terminal sorting domain-containing protein OS=Tannerella sp. oral taxon BU063 isolate Cell 6/7/9	116	ART	NC	C-terminal sorting domain-containing protein of the secretion system
W2CY98	Secretion system C-terminal sorting domain-containing protein OS=Tannerella sp. oral taxon BU063 isolate Cell 8/11	114	ART	NC	C-terminal sorting domain-containing protein of the secretion system
D7JDQ2	Transcriptional regulator_ XRE family OS=Bacteroidetes oral taxon 274 str. F0058	605	ART	NC	DNA binding

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[TFN1] *Protein function are based on protein ID from UniProt protein database (http://wwwwww.uniprot.org/); ^**Proteins with significantly altered expression are organized according to the ratio. ^*Indicates unique proteins in alphabetical order. Proteins in bold are increased more than twofold.