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. 2020 Dec 2;10:537650. doi: 10.3389/fcimb.2020.537650

Table 2.

Expression and activity of key HPV-related transcription factors in normal and tumor tissues of H&N region.

Transcription factor (family members) Specimen Transcription factor Activation status Expression level of transcription factor family members HPV-specific information References
AP-1
[c-Jun (39 kDa), JunB (36 kDa), JunD (35 kDa) c-Fos (41 kDa), FosB (36 kDa), Fra-1 (29 kDa), Fra-2 (35 kDa)]
HNSCC tumors (n = 7) and adjacent control tissues Increased binding of AP-1 in HIV LTR Nil Nil (Zoumpourlis et al., 1994)
HNSCC cell lines UM-SCC-1, -9, -11A, -11B, and-38 Active AP-1 with c-Jun, JunB, JunD and Fra-1 as major participants;
Loss of AP-1 site reduced IL-8 promoter activity
Nil Nil (Ondrey et al., 1999)
HNSCC cell lines UM-SCC-9 and -11B IL-1α-induced activation of AP-1;
Loss of AP-1 site moderately reduced IL-8 promoter activity
Nil Nil (Wolf et al., 2001)
HPV16 E7 transgenic mouse model with dominant negative c-Jun under human K14 promoter c-Jun associated AP-1 activation lost;
Prevention of chemical-induced skin papillomas
NIL Increased HPV16 E7 expression in DN-c-Jun mice; more papillomas (Young et al., 2002)
HNSCC cell lines UM-SCC-9 and -11B EGFR promoted AP-1 activity and downstream expression of VEGF Nil Nil (Bancroft et al., 2002)
HPV immortalized human oral keratinocytes- HOK-16B; Acetaldehyde activates AP-1 containing c-Jun Increased c-jun mRNA and protein accumulation HPV-transformed cells (Timmons et al., 2002)
HNSCC cell lines: UM-SCC 1, 5, 6, 9,11A, 11B, 22A, 22B, 38, and 46 Heterogeneity in cell lines with respect to AP-1 regulated gene expression Nil Nil, (Cell lines differed in their p53 expression (Yan et al., 2007)
FFPE-tissues of oral leukoplakias, with different degrees of epithelial dysplasia, and OSCC (n = 50); (HPV positivity-24%) According to degree of oral dysplasia withing lesion c-Jun nuclear expression increased and greatest expression and nuclear localization in OSCC Increased expression of c-Jun with increasing severity of the lesion Malignant progression mediated by c-Jun is independent of the presence of HPV in oral carcinogenesis (Acay et al., 2008)
Fresh oral tissue specimens (n = 100) Increased activation of AP-1 with increasing disease severity Presence of JunD with c-Fos in AP-1 complex in majority of oral cancer; JunB showed sporadic increase in a subgroup Nil (Mishra et al., 2010)
HPV immortalized human oral keratinocytes- HOK-16B
HNSCC cell line- UM-SCC-9, 11A, 11B, 38
Tobacco-carcinogen induced AP-1 reporter gene activity;
AP-1 activity dependent on c-Fos expression
c-Fos expression induced by tobacco carcinogens IL-8 and VEGF expressed both in HPV-transformed oral keratinocytes and HNSCC cell lines (Swenson et al., 2011)
Fresh tongue tissue biopsies (n-100), (HPV positivity-28%)
Tongue cancer cell lines-HPV16 positive- UPCI:SCC090; HPV negative- AW13516
Increased activation of AP-1 with increasing disease severity; Upregulation of downstream target genes: cyclin D1, c-myc, Bcl-xl, MMP-9, EGFR
Composition: c-Jun, c-Fos, Fra-2 in HPV-negative and JunD, JunB, c-Fos and Fra-2 in HPV-positive tumors
Higher expression of all Jun and Fos family members except Fra-1 which showed reciprocal kinetics Selective participation of JunD and JunB only in HPV16 positive tumors and cell lines; absence of c-Jun in HPV-positive (Gupta et al., 2015)
HNSCC cell line: HPV-positive- 93VU-147T Inhibition of AP-1 activation and change in active AP-1 by curcumin Curcumin induced loss of AP-1 c-Jun, JunD and JunB in HPV-positive oral cancer cells Nil (Mishra et al., 2015)
HNSCC cell lines – 26; Mouse model of metastasis Nil JunB, Fos, Fra-1, JunD overexpressed in cells with metastatic potential Nil (Hyakusoku et al., 2016)
Prospectively collected fresh biopsies - 116 and FFPE -30 from OSCC/OPSCC;
Total tissue analyzed (n = 116), FFPE (n = 66)
Nil Overexpression of JunD, and c-Fos in SCC as compared to normal; Overexpression of JunB only in HPV-positive tissues Differential increase in JunB in HPV-positive; Level of JunD upregulated but not differentially active only
HPV-positive cancers
(Verma et al., 2017)
Human OSCC samples (n = 123) Elevated expression of active AP-1 component phospho-c-Jun in resistant tumors; Active AP-1 strongly correlated with bcl-2 overexpression c-Jun overexpression in chemo-radioresistant tumors Nil (Alam et al., 2017)
HNSCC cell lines: HPV-negative- CAL33Res; HPV-positive- UM-SCC-47 Overexpression of c-Jun target gene AXL Silencing of c-JUN and c-FOS expression downregulated AXL expression and enhanced the sensitivity of HPV negative cells Targeting AP-1 enhanced the antitumor efficacy of BYL719 against HPV positive HNSCC (Badarni et al., 2019)
NF-κB
[NF-κB1 (50 kDa), NF-κB2 (52 kDa) RelA (65 kDa), RelB (70 kDa) c-Rel (78 kDa)]
HNSCC cell line: UM-SCC-9, -11B, and -38 Constitutive and inducible NF-κB, and promoter activity Overexpression of p65 in cell lines Nil (Duffey et al., 1999)
Human HNSCC cell lines: UM-SCC-1, 9, 11A, 11B, 38 Constitutive DNA binding & promoter activity; Constituents p65/Rel A and p50; IL-8 induction: stronger role than AP-1 Nil Nil (Ondrey et al., 1999)
Laryngeal SCC: Tumor and non-tumor laryngeal tissues Nuclear positivity of p65 High levels of p65 in cytoplasm, moderate in nucleus Nuclear p65 correlated with HPV16 E7 level (Du et al., 2003)
Oral tissue biopsies (n = 110).
OSCC (n = 66) OPMD (n = 34), normal (n = 10); HPV positivity = 22%, HR-HPV16 = 18%
Constitutively active NF-κB; increased with disease severity; Major participants:p50, p65, p52, c-Rel, RelB, and Bcl-3 Upregulation of p50, p65 and c-Rel with increasing severity of lesion; immunoreactivity for p52, c-Rel and RelB in cancer tissues. p50/p50 homodimerization common; Involvement of p65 only in NF-κB complex of HPV16-positive (Mishra et al., 2006)
HNSCC (n = 195); Control (n = 63; non cancer affected patients); Human HNSCC cell lines: Ho1N1, HSC2,and SKN3 SiRNA mediated downregulation of NF-κB activity; NF-κB nuclear staining: 55.6% HPV-negatives (above median-12.85), HPV-positive below median Overexpression of RELA, NF-κB1 Differential gene signatures.
Strong NF-κB activation in HPV-negative HNSCC
(Gaykalova et al., 2015)
OC and OPSCC Tissue biopsies (Fresh, n = 116; and
FFPE, n = 66)
Nil Variable presence of p50 and p65 in HPV+ and HPV- tissues p50: equal distribution among HPV-positive and HPV-negative tumors,
p65: strong correlation with HPV-postive tumors
(Verma et al., 2017)
Tongue tissue biopsies (n = 100)
TSCC cell line: UPCI:SCC090 and AW13516
Increase d activity with severity of disease; p50 and c-Rel forming NF-κB Differential expression of NF-κB proteins Selective participation of p65 in NF-κB complex of HPV16-positive HNSCC (Gupta et al., 2018)
STAT3 (88 kDa) HNSCC Cell line:YCU-N861, YCU-H891; HNSCC tissues (n = 6) Constitutive activation of
EGFR and STAT3; STAT3DN66 and STAT3DN99 abrogate STAT3 activity
Constitutive expression of STAT3 Nil (Masuda et al., 2002)
HNSCC tissues (n = 90) High level of pSTAT3 in early stages (T1, T2), moderate in late stages (T3, T4); No STAT3 in normal samples HNSCC - 82% with high or intermediate STAT3; No STAT3 in normal tssues Nil (Nagpal et al., 2002)
HNSCC cell lines: HN6, HN12, HN13, HN30, HaCaT, HEK293T, HEK293FT cells; HNSCC FFPE tissues (n = 460) pSTAT3 induction by IL-6 produced through active NF-κB signalling STAT3 expression unaffected Nil (Squarize et al., 2006)
HNSCC (n = 195); Control (n = 63; non cancer affected patients); Human HNSCC cell lines: Ho1N1, HSC2,and SKN3 Transcript profiling of STAT3 with NF-κB pathway signature target genes (IRF1, CEBPD, CCND1, ICAM1, JAG1, JAK3, and NOS3); reporter gene expression Indirect evidence Specific signature for HPV positive and negative; 49 HPV negative, while 1 HPV positive patients stained for nuclear STAT3 above median (Gaykalova et al., 2015)
OC and OPSCC Tissue biopsies (Fresh, n = 116; and
FFPE, n = 66)
pSTAT3Y expression Immunoblotting and Immunohistochemistry: inverse correlation between HPV positivity and STAT3/pSTAT3 expression. HPV positive: low STAT3/pSTAT3; HPV negative: high STAT3/pSTAT3 (Verma et al., 2017)
SOX2
(34 kDa)
OSCC frozen tumor tissue samples (n = 40) SOX2 overexpression associated with increased activity High expression of SOX2 and CyclinE1 in OSCC specimens Nil (Freier et al., 2010)
HNSCC tumor tissues (n = 496) SOX2 induced BCL-2 and enhanced chemo-resistance SOX2 amplification in HPV-negative, but no amplification in HPV-positive (Schrock et al., 2014)
H&N tissue retrospectively collected (n = 94) Nuclear positivity of SOX2 SOX2 expression detected in 95% of laryngeal dysplasia Nil (Granda-Diaz et al., 2019)
OPSCC (n = 157) Nil SOX2 overexpression with poor prognosis Poorer overall survival in SOX2-amplified HPV-negative cases (p = 0.036) (Dogan et al., 2019)
Tongue SCC cell line- CAL‐27 EGFR mediated stabilization and upregulation of SOX2 expression Nil (Lv et al., 2020)
YY1 (44 kDa) Laryngeal SCC Cell line - Hep-2; Control- HEK293T Proliferation and migration with suppression of apoptosis as indicators of increased YY1 activity YY1 upregulated in LSCC Nil (Qu et al., 2017)
NPC cases (n = 40); Stage I replication (297 cases and 611 controls); Stage II replication (768 cases and 1526 controls) YY1-mediated repression of TRIM26 Nil Nil (Lyu et al., 2018)
OSCC tumor tissues (n-30); Cell lines- HEK293T, AW8507 YY1 mediated reporter gene activation by CARM1 mediated arginine methylation YY1 overexpressed in oral cancer Nil (Behera et al., 2019)
SP1 (80.6 kDa) Normal oral mucosa (n-8), OSCC (n-10); OSCC cell lines: SCC-15, YD-15 SP-1 inhibitor downregulated expression of SP-1 and decrease tumor growth SP-1 was overexpressed in oral tumors compared to normal Nil (Shin et al., 2013)
NPC tumors (n-82), Metastatic malignant tumors (n-60), Nonmetastatic malignant tumors (n-22) miR-24 overexpression lead to reduced SP1 activity and inhibited proliferation Reduced SP1 expression contributed to the reduction in radio-resistance Nil (Kang et al., 2016)
OSCC patients (n-55);
Cell line- CAL-27 and SCC9
Upregulated SP-1 in OSCC SP-1 is overexpressed in OSCC and could promote cell invasion and migration in OSCC Nil (Liu et al., 2019)
OCT-1 (76.4 kDa) HNSCC cell line: PCI-04A Ionizing radiation induce OCT-1 Nil (Meighan-Mantha et al., 1999)
c/EBPβ
(76.4kDa)
HNSCC cell lines: UM-SCC-1, -9, -11A, -11B, and -38 Constitutive activation Nil (Ondrey et al., 1999)
NPC and non-cancerous NPE (n-33); Cell line- HNE1 and 5–8F Nil (Wang et al., 2016)
FOXA1 (49.1 kDa)/FOXA2 (48.3 kDa) HNSCC (n = 152) Nil FOXA1 expressed in basal cells of squamous epithelium, pre-invasive HNSCC lesions Due to specificity of FOXA-1 positive site, it may have implication in HPV-mediated pathogenesis (Karpathiou et al., 2017)
NPC tissues (n-114), Non-cancer inflammatory NPE tissues (n-64) FOXA1 regulated TGF-β-stimulated transcriptome FOXA1 protein was decreased in NPC cells; loss of FOXA1 associated with lymph node metastasis and poor prognosis. Nil (Li et al., 2019)

AP-1, Activator Protein; DMBA, 7,12-dimethylbenz[a]anthracene; DN, Dominant Negative; EGFR, Epidermal Growth Factor Receptor; FFPE, Formalin-Fixed and Paraffin-Embedded; FOXA1, Forkhead Box A1; FOXA2, Forkhead Box A2; HPV, Human Papillomavirus; HNSCC, Head and Neck Squamous Cell Carcinoma; LSCC, Laryngeal Squamous Cell Carcinoma; NF-κB, Nuclear Factor-κB; NPC, Nasopharyngeal Carcinoma; NPE, Nasopharyngeal Epithelial; STAT3, Signal Transducer and Activator of Transcription – 3; SOX2, sex determining region Y (SRY)-box; SP1, Specificity Protein 1; TSCC, Tongue Squamous Cell Carcinoma; TPA, 12-O-tetradecanoylphorbol-13-acetate; OCT-1, Octamer Transcription Factor-1; OC- Oral Cancer; OSCC, Oral squamous Cell Carcinoma; OPSCC, Oropharyngeal Squamous Cell Carcinoma; OPMD, oral premalignant disease; VEGF, Vascular Endothelial Growth Factor; YY1, Yin Yang 1.