. 2021 Jul 20;12:699044. doi: 10.3389/fphar.2021.699044

TABLE 4.

Preclinical studies showing therapeutic phytochemicals/herbal derivatives against tobacco/alcohol-associated HNC.

Bioactive compound/ Herbal derivative	Cell type/Model	Test and oosage	Anti-tumour outcome	Molecular outcome	References
PubChem CID (Class)
Source
Curcumin	In vitro: MDA 1986 (OSCC), Tu 686 (LSCC), Tu 167, JMAR C42 (Floor of mouth OSCC), MDA 686LN (OPSCC)	Cell proliferation: 10 μM, 50 μM	• Proliferation↓	• NF-κB activation↓	Aggarwal et al. (2004)
CID- 969516			• Apoptosis↑	• Bcl-2↓, cyclin D1↓, IL-6↓, COX-2↓, MMP-9↓
(Phenolics)			• Arrests cell cycle in G1/S phase
Curcuma Longa			• Arrests cell cycle in G1/S phase
Gossypol	In vitro: UMSCC-1, UMSCC-17B (Floor of mouth OSCC), Human oral keratinocytes and Normal keratinocytes	Cell proliferation:	• Proliferation↓	NA	Wolter et al. (2006)
CID- 3503		UMSCC-1, UMSCC-17B, Human oral keratinocytes and Normal keratinocytes IC50- 3, 6,2, 12.5 μM respectively	• Growth of tumour↓
Phenolic			• Mitotic rate↓
Gossypium arboreum L.	In vivo: NCr-nu/nu mice		• Apoptosis↑
Berberine	In vitro: HSC-3 (Tongue OSCC)	Cell viability:	• Cell viability↓	• Bcl-2↓, BAX↑, p53↑	Lin et al. (2007)
CID: 2353		10 μM	• G0/G1-phase arrest	• Cyt C release
(Alkaloid)			• ROS↑,Ca²⁺↑, MMP↓, Apoptosis↑
Rhizoma coptidis
	In-vitro study: FADU (Hypopharyngeal SCC)	Cell cytotoxicity:	• Cytotoxicity ↑	• FasL ↑, TRAIL ↑	Seo et al. (2015)
		Cell cytotoxicity:	• Cytotoxicity ↑	• Cleaved caspase-8 ↑, cleaved
		12 or 25 μM for 24 h	• Apoptosis ↑	• caspase-7 ↑
			• Cell viability ↓	• Bcl-2 ↓, Bcl-xL ↓, Bax ↑, Bad ↑, Apaf-1↑, cleaved caspase-9 ↑, cleaved caspase-3 ↑, PARP ↑
			• Cell migration ↓
				• MMP-2 ↓, MMP-9 ↓
				• ERK, JNK and p38 phosphorylation ↓
Physalis angulate	In-vitro study: HSC-3 (OSCC)	Cell viability:	• Mitochondrial reductase activity ↓	• ROS ↑	Lee et al. (2009)
Physalis angulate		Cell viability:	• Apoptosis ↑	• Bcl2 ↓, Bax ↓, AIF ↓, cytochrome c ↓, proform caspase-3 protein levels ↓, caspase-9 ↑, proform caspace-4 protein levels ↑, MMP attenuated
(Crude extract)		IC50: 10 μg/ml	• Oxidative stress ↑
			• Loss of cell function ↑
			• Loss of cell function ↑	• ORP150 ↓, HSP70 ↑
			• S- and G2/M-phase arrest	• HO-1 ↑, SOD ↑
			• Mitochondrial function impaired	• HO-1 ↑, SOD ↑
Guggulsterone	In vitro: SCC-4 (Tongue OSCC), HSC-2 (OSCC)	Protein expression:	NA	• Phosphorylation of p65, IkBα & STAT3, NF-κB↓
CID- 6450278		50 μM
(3-hydroxy steroid)				• IL-6↓ , COX-2↓
Commiphora mukul				• Expression VEGF↓
Proanthocyanidins	In vitro: UMSCC-1, UMSCC-5 (Floor of mouth OSCC), FaDu (Hypopharyngeal OSCC), OSC-19 (Tongue OSCC), Beas-2B (Bronchial Epithelium transformed with Ad12-SV40 2B)	In vitro: IC50- 101 nM, 67 nM for	• Cell viability↓	• Cyclin D1↓ and Cyclin D2↓	Prasad and Katiyar, (2012)
CID- 108065		24 and 48 hrs, respectively	• Apoptosis↑	• cdk2↓, cdk4↓, cdk6↓
		24 and 48 hrs, respectively	• G1 phase arrest	• Cip1/p21↑ and Kip1/p27↑
		In vivo: 0.5%, w/w	• 61% less tumor volume (p<0.001)	• Bax↑, Bcl-2↓, caspase 3↑ and poly(ADP-ribose)polymerase↑.
(Phenolic)
Vitis vinifera
Vitis vinifera	In vivo: athymic nude mice
Wogonin	In vitro: NPC-TW076, NPCTW039 (NPSCC)	Autophagy: 50 μM	• Autophagy↑	• P13K/Akt↓, mTOR/P70S6K↓, c-Raf/ERK↓	Chow et al. (2012)
CID- 5281703			• Apoptosis↑
(Phenolic)
Scutellaria baicalensis
Caffeic Acid Phenethyl Ester	In vitro: TW2.6 (OSCC)	Cell proliferation: IC50- 83.8, 46.6, and 18.8 μM for 24, 48, and 96 h treatment, respectively	• Cell proliferation↓, colony formation↓	• Akt↓, GSK3β↓, FOXO1↓, FOXO3a↓, NF-κB↓, Rb↓, Skp2↓, cyclin D1↓, p27Kip↑	Kuo et al. (2013)
CID- 5281703			• Apoptosis↑
(Phenolic)			• G1 phase↓, G2/M phase↑ cell population
Populus nigra L.			• G1 phase↓, G2/M phase↑ cell population
Ellagic acid	In vitro: HSC-2 (OSCC), HF-1 (Normal fibroblasts)	Cytotoxicity: (IC50- 260 & 142 μM on 2^nd and 3^rd day of exposure, respectively)	• Apoptosis↑	• Caspase 3/7 ↑	Weisburg et al. (2013)
CID- 5281855				• Cleavage of poly ADP ribose polymerase
(Phenolic)
Rubus occidentalis
Cucurbitacin	In vitro:	Cytotoxicity: IC50- 3.7 μM	• Sub G0/G1 phase arrest	• Caspase 3 Activation	Hung et al. (2013)
CID- 5281316	SAS (Tongue OSCC)		• Apoptosis↑	• MMP↓
(Terpene)
Cucumic melo L
BME	In vitro: Cal-27 (Tongue OSCC), JHU-22, JHU-29 (Laryngeal SCC)	In vitro: 1% BME	• Cell proliferation↓	• c-Met signaling pathway↓	Rajamoorthi et al. (2013)
(crude extract)		In vivo: 100 μl BME (0.1 g/ml) for 5 days	• Tumor growth and volume↓	• Mcl-1, pSTAT3, cMyc expression↓
Momordica charantia			• Keratinocyte formation↓, mitosis↓	• cyclinD1↓, survivin↓, cell arrest
Momordica charantia	In vivo: BALB/c athymic nude mice		• Keratinocyte formation↓, mitosis↓	• MCM2↓
Honokiol	In vitro: SCC-1, SCC- 5 (Floor of mouth OSCC), OSC-19 (Tongue OSCC), FaDu (Hypopharyngeal SCC)	In vitro: ∼60 μM for 24, 48 and 72 hrs	• Cell viability↓	• Expression levels of Cyclins: D1, D2, and Cdks: 4 and 6↓	Singh et al. (2015)
CID-72303		In vitro: ∼60 μM for 24, 48 and 72 hrs	• Apoptosis↑	• Expression levels of Cyclins: D1, D2, and Cdks: 4 and 6↓
(Phenolic)		In vivo: 100 mg/kg body weight (mw-266.3)		• EGFR↓, mTOR and their downstream signalling molecules↓
Magnolia officinalis		In vivo: 100 mg/kg body weight (mw-266.3)		• EGFR↓, mTOR and their downstream signalling molecules↓
Cepharanthine	In vitro: CNE-1, CNE-2 (NPSCC)	Cell proliferation:	• Cell proliferation↓	• DNA repair genes ↓	Liu et al. (2015)
CID- 10206		IC50 for CNE-1 and CNE-2: 20 and 32 nM after 48 hrs, respectively	• G1 phase arrest
(Alkaloid)
Stephania cepharantha
MEAG	In vitro: MC3 (Chronic myelogenous leukemia), HN22 (OSCC)	Cytotoxic effect:	• Mitochondria mediated apoptosis↑	• MMP↓, Cyt C release, caspase 9 ↑, t-Bid↑, cleaved caspase-8↑, DR5↑.	Lee et al. (2016)
(Crude extract)		IC50 for MC3 and HN22: 6.5 and 4.6 μg/ml, respectively	• Mitochondria mediated apoptosis↑
(Terpenes)			• Nuclear condensation and fragmentation
Withania somnifera			• Nuclear condensation and fragmentation
Goniothalamin	In vitro: H400 (OSCC)	Cytotoxic effect:	• Cell viability↓	• MMP↓, Cyt C release	Li et al. (2016)
CID- 6440856		IC50: 8.9 nM after 72 h	• Cell proliferation↓	• NF-κB activation↓
(Phenolic)			• Apoptotic-like morphology (cell shrinkage, dense cytoplasm, blebbing of cell surface)
Goniothalamus marcrophyllus
Goniothalamus marcrophyllus			• S phase arrest.
Lupeol	In vitro:	Cell viability:	• G1 phase arrest	• Expression p53↑, Bax↑, CDKN2A↑, CyclinD1↓, Ki67↓	Bhattacharyya et al. (2017)
CID- 259846	HEp-2 (Human papillomavirus-related endocervical adenocarcinoma), UPCI:SCC131 (Floor OSCC)	IC50 for Hep-2 and SCC131: 53.5 and 52.4 μM after 24 hrs, respectively	• Apoptosis↑ via intrinsic pathway	• Expression p53↑, Bax↑, CDKN2A↑, CyclinD1↓, Ki67↓
(Terpene)			• Cell viability↓	• Caspase 3 activation
Camellia japonica
Camellia japonica	Ex vivo: Fresh HNSCC tumor tissues
Icaritin	In vitro: KB (Human papillomavirus-related endocervical adenocarcinoma), SCC9 (Tongue OSCC)	Cell viability:	• Mitochondria mediated apoptosis↑	• MMP↓, Cyt c release	Jin et al. (2017)
CID- 5318980		∼20 μM for 24 and 48 h		• miR-124↑
(Phenolic)				• Sp1/DNMT1 signaling↓
Epimedium grandiflorum
Osmuda regalis root	In vitro: FaDu (Hypopharyngeal SCC), HLaC78, HLaC79, HLaC79-tax (LSCC)	Cytotoxicity:	• Cell growth↓	• CLEC3B↓, KAL1↓, MMP 11↓, MMP 15↓, MMP2↓	Schmidt et al. (2017)
(crude extract)		IC50 for HLaC79, FaDu, HLaC79-Tax and HLaC79: 21.4, 8.5, 20.6 and 9.9 μg/ml, respectively	• Apoptosis↑	• CLEC3B↓, KAL1↓, MMP 11↓, MMP 15↓, MMP2↓
Osmuda regalis			• Invasion↓	• Integrin such as ITGB3↑, ITGA1↑, ITGAM↑
Osmuda regalis			• Invasion↓	• metastasis genes such as CTFG↑, PPIA↑, SELP↑, VCAN↑
Piperine	In vitro: KB (Human papillomavirus-related endocervical adenocarcinoma)	Cell proliferation:	• Apoptosis↑	• ROS↑, MMP↓	Siddiqui et al. (2017)
CID- 638024		IC50: 124 μM for 24 h	• Cell viability↓	• Caspase 3 activation
(Alkaloid)			• Cell growth↓
Piper nigrum			• Cellular morphological changes
Piper nigrum			• G2/M phase arrest
Embelin	In vitro: SCC25, Cas9-22, YD10B (Tongue squamous cell carcinoma)	Cytotoxicity:	• Cell viability↓	• MMP↓, caspase 9 & 3↓	Lee et al. (2017)
		0–300 μM for 24 h; cell specific effect	• Condensed fragmented nuclei	• p62/SQSTM1↓
			• Autophagic vacuoles appears	• Conversion↑ LC3-I to LC3-II
CID- 32798
(Phenolic)
Embelia ribes
Dihydroartemisinin	In vitro: Cal-27 (Tongue OSCC)	Cell proliferation:	• Autophagosomes↑	• LC3-II ↑, Bevlin-1↑, γH2AX foci↑	Shi et al. (2017)
CID- 3000518		IC50: 24.4 μM for 24 h	• Oxidative stress↑	• STAT3 activation↓ and disrupted p-STAT3 nuclear translocation
CID- 3000518			• DNA double-strand break
(Sesquiterpene lactone)			• Weight and volume of Xenograft tumor↓ by 56.58%
Artemisia annua			• Weight and volume of Xenograft tumor↓ by 56.58%
β-Elemene	In vitro: YD-38 (Gingival OSCC)	Colony formation and Apoptosis:	• Cell proliferation↓	• Expressions of STAT3↓, p-STAT3↓, p-JAK2↓, and Bcl-2↓, Bax↑ and caspase-3↑	Huang and Yu, (2017)
CID- 6918391	In vivo: BALB/c nude mice	Colony formation and Apoptosis:	• Apoptosis↑
(Terpene)		195 nM β-Elemene
Curcuma wenyujin		195 nM β-Elemene		• Block JAK2-STAT3 pathway
Thymol	In vitro: Cal27, SCC-4, SCC-9 (Tongue OSCC)	Cytotoxicity:	• Cell viability↓	• c-PARP↑, MMP dysfunction	De La Chapa et al. (2018)
CID- 6989	In vivo: Athymic nu/nu mice	2.3 µM	• No colony formation
(Terpene)			• In vivo tumor growth↓
Thymus vulgaris			• Apoptosis↑
Tanshinone	In vitro: SCC-9 (Tongue OSCC)	Cell viability:	• Apoptosis↑	• LC3-I↑	Qiu et al. (2018)
CID- 114917	In vivo: BALB/c-nu	IC50: 17.5 μM.	• Autophagy↑	• Beclin-1/Atg7/Atg12-Atg5 pathway↑
(Abietane diterpenoid)			• Growth of solid tumors in vivo	• PI3K/Akt/mTOR pathway↓
Salvia miltiorrhiza			• Growth of solid tumors in vivo	• PI3K/Akt/mTOR pathway↓
Oridonin	In vitro: UM1, SCC25 (Tongue OSCC)	Cell proliferation:	• G2/M phase arrest	• Bax/Bcl-2↑	Yang et al. (2018b)
CID- 5321010		IC50 for SCC25, UM1, UM2, HSC3 and Ca127: 9.1, 8.2, 10.6, 15.4 and 9.6 μM, respectively.	• Apoptosis↑	• Cyclin B1↓, pCDK1↑, cyclin D1↑, cyclin D3↑, p21↑ and cyclin A2↑
(Terpene)
Rabdosia rubescens				• Activates caspase-3, caspase-9 and PARP-1
Rabdosia rubescens				• P13K/Akt/mTOR pathway↓
Epigallocatechin-3-gallate	In vitro: HSC (Tongue OSCC)	Cell proliferation:	• Cell viability↓	• Caspase‑3 and -7↑	Yoshimura et al. (2019)
Epigallocatechin-3-gallate	In vivo: BALB/c nude (nu/nu) mice	IC50 value at 24, 48 and 72 h were >100, 43.2 and 39.3 μM, respectively	• G1 phase arrest	• miR-22↑
CID- 65064			• Apoptosis↑
(Phenolic)			• Tumor size↓ 45.2%
Camellia sinensis			• Tumor size↓ 45.2%
Quercetin	In vitro: hNOK (Human normal oral keratinocytes), Tc8113, SAS (Tongue OSCC)	Cell viability:	• Cell viability↓	• miR-22/WNT1/Beta-catenin pathway↓	Zhang et al. (2019)
(Phenolic)		CC₅₀ for hNOK, Tc8113 & SAS: 298.6, 48.7 & 44.3 mM, respectively	• Tumor volume and weight↓
CID: 5280343
Allium cepa L.
	In vivo: BALB/c nu/nu mice
Ursolic Acid	In vitro: Ca9-22 (Tongue OSCC), SCC2095 (OSCC)	Cell proliferation:	• Caspase-dependent apoptosis	• Akt/mTOR/NF-κB signaling↓, ERK↓, and p38↓	Lin et al. (2019)
CID- 64945		IC50 for UA, Ca9-22: 11.5 and 13.8 μM, respectively	• Autophagy↑, autophagosomes↑	• Akt/mTOR/NF-κB signaling↓, ERK↓, and p38↓
(Terpene)			• Migration↓, Invasion↓	• LC3B-II conversion, p62↑
Salvia rosmarinus			• Migration↓, Invasion↓	• Proteolytic activity of MMP-2↓
Shikonin	In vitro: 5-8F (NPSCC)	Cell proliferation:	• Necroptosis↑	• Necrostatin-1↑	Liu et al. (2019)
CID- 479503	In vivo: BALB/c nude mice	IC50: 7.5 μM after 6 h	• In vivo tumor growth↓	• RIPK1↑, RIPK3↑, MLKL↑
(Hydroxy-1,4-naphthoquinone)				• Caspase-8 and -3↑
(Hydroxy-1,4-naphthoquinone)				• ROS↑
Lithospermum erythrorhizon				• ROS↑
Chrysophanol	In vitro: FaDu (Hypopharyngeal SCC), SAS (Tongue OSCC)	Cell viability:	• Cell viability↓	• ROS↑	Hsu et al. (2020)
CID- 10208		IC50 for FaDu and SAS: 9.6 ± 1.3 and 12.6 ± 2.1 μm at 24 h, respectively	• G1 phase arrest	• Expression of procaspase 3↓, cyclin D1↓, CDK4↓, CDK2↓, cdc2↓
(Phenolic)			• Metastasis↓, EMT↓
Rheum rhabarbarum			• Metastasis↓, EMT↓
Moscatilin	In vitro: FaDu (Hypopharyngeal SCC)	Cell cytotoxicity:	• Cell viability↓	• Activation of caspases-3,-8,-9,-7↑	Lee et al. (2020)
CID- 176096		IC50: 1.4 μM at 72 h	• Cell proliferation↓	• MMP↓, Cyt C release
(Phenolic)			• Apoptosis via intrinsic as well as extrinsic pathway	• JNK pathway↓
Dendrobium sp.			• Apoptosis via intrinsic as well as extrinsic pathway	• JNK pathway↓
Demethoxycurcumin	In vitro:	Cell proliferation:	• Cell viability↓	• cIAP1/XIAP↓, heme oxygenase-1↑	Chien et al. (2020)
Demethoxycurcumin	SCC-9, HSC-3 (Tongue SCC)	IC50: 50 μM	• Cell proliferation↓	• Caspase-3↑, -9↑, -8↑, p38-MAPK-HO-1 signaling↑, MAPK↑, JNK1/2↑
CID- 5469424			• G2/M phase arrest
(Phenolic)			• Morphological changes
Curcuma Longa			• Morphological changes

Abbreviations: Akt- Protein kinase B; Apaf-1- Apoptotic protease activating factor 1; Atg7- Autophagy related 7; Bad- BCL2-associated Agonist of cell Death; Bcl-2- B-cell lymphoma 2; BME- Bitter Melon Extract; Cdc2- Cell division control 2; Cdk2- Cyclin-dependent kinase 2; CDKN2A- Cyclin-dependent kinase inhibitor 2A; cIAP- Calf Intestinal Alkaline phosphatase; Cip1- CDK interacting protein 1; CLEC3B- C- type lectin domain family three member B; c-Met- tyrosine-protein kinase Met; COX-2- Cyclooxygenase-2; c-PARP- Cleaved Poly (ADP-ribose) polymerase; c-Raf-c- Rapidly Accelerated Fibrosarcoma; CTGF- Connective tissue growth factor; Cyt C- Cytochrome complex; DNMT1- DNA (cytosine-5)-methyltransferase 1; DR5- Death receptor five; ERK- Extracellular-signal-regulated kinase; FasL- Fast ligand or cell death receptor; FOXO1- Forkhead box protein O1; GSK3β- Glycogen synthase kinase three beta; H&N- Head and neck; HO-1- Heme oxygenase 1; HSP70–70 kilodalton heat shock proteins; IkBα- I-kappa-B-alpha; ITGA1- Integrin alpha-1; ITGAM- Integrin alpha M; ITGB3- Integrin beta three; JNK- c-Jun N-terminal kinase; Kip1- kinase inhibitor 1; LC3- Microtubule-associated protein 1A/1B-light chain three; MAPK- Mitogen-activated protein kinase; Mcl-1- Myeloid leukemia cell differentiation protein 1; MCM2- Minichromosome maintenance protein complex 2; MLKL- Mixed lineage kinase domain-like pseudokinase; MMP- Matrix metallopeptidase; mTOR-mammalian target of rapamycin; NF-κB- Nuclear factor kappa light chain enhancer of activated B cells; ORP150–150-kDa oxygen-regulated protein; P13K- Phosphatidylinositol 3-kinase; P70S6K- 70-kDa ribosomal protein S6 kinase; PARP- Poly-ADP ribose polymerase; JAK- Janus kinase; PPIA- Peptidylprolyl isomerase A; Rb- Retinoblastoma protein; RIPK1- Receptor-interacting serine/threonine-protein kinase 1; ROS- Reactive oxygen species; SOD- Superoxide dismutases; Sp1- Specificity protein 1; SQSTM1- Sequestosome-1; STAT3- Signal transducer and activator of transcription three; t-Bid-truncated BH3 interacting-domain death agonist; TRAIL- TNF-related apoptosis-inducing ligand; VEGF- Vascular endothelial growth factor; WNT1- Wingless-related integration site 1; XIAP- X-linked inhibitor of apoptosis protein; yH2AX- Phosphorylated X-linked inhibitor of apoptosis protein.