J Food Bioact

Journal of Food Bioactives, ISSN 2637-8752 print, 2637-8779 online

Journal website www.isnff-jfb.com

Review

Volume 7, September 2019, pages 18-26

Anticancer and antiproliferative properties of food-derived protein hydrolysates and peptides

**Table 1.** Sources, methods of production and mechanisms of action of anticancer food-derived protein hydrolysates and peptides*
Source	Protein hydrolysates/Peptide	Production Method	Cellular and/or animal model	IC₅₀ value	Mechanism	References
*Cell lines Key: MCF-6, MCF-7, MDA-MB-231, BT549–breast cancer; HeLa–cervical cancer; Caco-2, HCT-116, TC7, DLD-1–colon cancer; DU-109–esophageal cancer; NIH3T3–mouse fibroblast; HT1080–fibrosarcoma; U87-glioma; L1210- lymphocytic leukemia; HepG-2–liver cancer; A549, DU-1299–lung cancer; U937–histiocytic lymphoma; HT-29, P388D1-mouse monocyte macrophage cell line; RAW264.7 (mouse)–macrophage; MC3T3E1–osteoblastic calvaria-derived cells; DU-145–prostate; UMR106–rat osteosarcoma-derived cells; AGS–stomach adenocarcinoma; HRA–human ovarian cancer; C33–epidermoid cervical carcinoma; B1F610-melanoma cells; SKVO-3–ovarian cancer cell; HUVEC–Human umbilical vein endothelial cells.
Tuna cooking juice	KPEGMDPPLSEPEDRRDGAAGPK, KLPPLLLAKLLMSGKLLAEPCTGR	Enzymatic hydrolysis with Protease XXIII and ultrafiltration	Human breast cancer MCF-7 cells and MCF-10A mammary epithelial cells	1.39 mg/mL	Apoptosis induction and tumour suppression	Hung et al., 2014
Peptides from Spirulina platensis	HVLSRAPR	Enzymatic hydrolysis and fractionation	HT-29 cancer cells	99.88 µg/mL	Tumour suppression	Wang and Zhang, 2017
Mung bean vicilin	Protein hydrolysates and vicilin seed extracts	Ammonium sulfate fractionation of vicilin followed by separate hydrolyses with alcalase and trypsin	MCF-7 and MDA-MB-231 breast cancer cells	0.32, 0.73 and 0.45 mg/mL for MCF-7; 0.26, 0.48 and 0.54 mg/ml for MDA-MB-231 cells	Tumour suppression	Gupta et al., 2018
Chickpea	Protein hydrolysates	Alcalase hydrolysis	Tumour-bearing mice	Not mentioned	Tumour suppression	Xue et al., 2012
Maize	Peptides	Alcalase hydrolysis followed by ion exchange column chromatography fractionation	Hepatocarcinoma human HepG2 cells	1.5–150 µg/mL for peptide fractions and 0.001–1.5 mM for derived peptides.	Apoptosis induction, cell growth inhibition	Ortiz-Martinez et al., 2017
Soybean (germinated)	Peptide fractions	Extraction, precipitation, enzymatic hydrolysis and membrane fractionation	HeLa cells	1.83 mg/mL	Apoptosis induction and down regulation of PTTG1 and TOP2A tumorigenic genes	Robles-Ramírez et al., 2012
Soybean	Lunasin	Simulated GIT digestion with pepsin and pancreatin	L1210 leukemia cells	14 µM	Activation of Caspase-3 and dose-dependent G2 cell cycle arrest	González de Mejia et al, 2010
Soybean	Lunasin	Lunasin obtained by aqueous extraction from soybean followed by ion exchange and size exclusion chromatography, and further purified by HPLC separation	HT-29 colon cancer cells	61.7 µM	Activation of mitochondrial pathways and increased induction of pro-apoptotic nuclear clusterin expression	Dia and Mejia, 2010
Soybean	XMLPSYSPY	Thermoase and ethanol extraction	P388D1	0.16 mg/mL	Not investigated	Kim et al., 2000
Soybean	Isoflavone-derived soy	Sequential hydrolysis with endopeptidase, exopeptidase and amylase in that order	MCF-7	Not stated	Apoptosis via p21, p53, and caspase-3 expression, and inhibition of VEGF and HSP90	Park et al., 2009
Soybean	Kunitz trypsin inhibitor		HRA human ovarian cancer cells	2–3 µM	Inhibition of the upregulation of urokinase-type plasminogen activator, uPA, and its specific receptor, uPAR	Kobayashi et al, 2004
Soybean	VFDGEL, IETWNPNNKP and FEITPEKNPQ	Pepsin and pancreatin	L1210	7.9, 4.0 and 2.4 mM in that order	Human topoisomerase II inhibition	Wang et al., 2008
Rapeseed	Rapeseed peptide	Alcalase and flavourzyme	HeLa	Not stated	DNA structural damage and apoptosis induction	Xue et al, 2010
Amaranthus mantegazzianus (seed)	Mantegazzianus peptide isolate	Alcalase	UMR106, Caco-2, TC7, MC3T3E1	1.0, 1.5 and 2.5 and 2.5 mg/mL respectively	Inhibition of cell adhesion, necrosis and apoptosis	Barrio and Añón, 2010
Walnut	Cys-Thr-Leu-Glu-Trp	Alkaline extraction, precipitation by centrifugation and hydrolysis with papain	Human HeLa cervical cancer and MCF-7 breast cancer cells	0.60 and 0.65 mg/mL respectively	Induction of apoptosis and autophagy	Ma et al., 2015
Tilapia	Tilapia hepcidin (TH) 1–5	Chemically synthesized	HeLa, HepG2, HT1080	Not stated	Downregulation of interleukin (IL)-6, IL-8, IL-12, IL-15, interferon-γ, Bcl-2 and caspase-7, as well as upregulation of IL-2 and IL-8	Chang et al, 2011
Estuary cod (Epinephelus coioides)	Epinecidin-1	Chemically synthesized	A549, HepG2, HT1080, U937, NIH3T3, RAW264.7 cancer cells	Not stated	Inhibition of cell growth and cell lysis	Lin et al., 2009
Cuttlefish ink (Sepia esculenta)	An oligopeptide, N Gln-Pro-Lys	Trypsin	DU-145 prostate cancer cells	Variable	Inhibition of cell viability and growth	Ding et al, 2011
Tuna dark muscle byproduct	LPHVLTPEAGAT PTAEGGVYMVT	Papain and protease XXIII	MCF-7	8.1 µM and 8.8 µM respectively	Inhibition of cell viability and proliferation	Hsu et al., 2011
Loach (Misgurnus anguillicaudatus)	Loach peptide fractions	Loach tissue homogenization followed by papain hydrolysis	HepG2, MCF-7 and Caco-2 cells	Not stated	Inhibition of cell proliferation	You et al., 2011
Setipinna taty	Heated products from peptic hydrolysates of marine fish (half-fin anchovy)	Peptic hydrolysates heated for 10 min at 95 °C	DU-145 prostate, 1299 lung, and 109 esophagus cancer cells	13.67, 25.17 and 40.28 mg/mL respectively	Cell proliferation inhibition	Song et al., 2011
Skate (Raja porosa) cartilage protein hydrolysate	FIMGPY	Enzymatic hydrolysis with pepsin, trypsin and chymotrypsin	HeLa	4.81 mg/mL	Apoptosis and tumour suppression	Pan et al, 2016
Blue whiting cod, Plaice, Atlantic Salmon	Fish protein hydrolysates	pH-shift extraction followed by Protamex and alcalase hydrolysis	MCF-7/6, MDA-MB-231 and breast cancer cells	Not stated	Inhibition of cell growth and proliferation	Picot et al., 2006
Rice bran	Alacalase hydrolysis followed by simulated GIT digestion	Gastrointestinal juice	Caco-2, HepG2, HCT-116, MCF-7, MDA-MB-231	Not mentioned	Not mentioned	Kannan et al., 2010
Snow crab (by-products)	KCl2 cationic peptide	Enzymatic hydrolysis with Protamex, electrodialysis, ultrafiltration	A549, HCT15, BT549, PC3	Not mentioned	Not mentioned	Doyen et al., 2011
Squid gelatin	Squid gelatin protein hydrolysates	Protamex, trypsin, Neutrase, Savinase, NS37005, Esperase, Alcalase	MCF-7, U87	130 and 100 µg/mL for MCF-7 and U87 respectively	Not stated	Alemán et al., 2011
Rough sea squirt/solitary tunicate	Solitary tunicate (Styela clava) enzymatic hydrolysates	Alcalase, Thermoase PC10F, pepsin,	AGS, DLD-1, HeLa	577.1–1,240.0 µg/mL	Not mentioned	Kim, 2011
Japanese threadfin bream	Nemipterus japonicus tryptic hydrolysates	4 h hydrolysis with trypsin at 37 °C	HepG2 liver cancer cells	61.1 µg/mL	Inhibition of cell viability	Naqash and Nazeer, 2012
Beef sarcoplasmic protein hydrolysates	Gly-Phe-His-Ile, Asp-Phe-His-Ile-Asn-Gly, Phe-His-Asp, and Gly-Leu-Ser-Asp-Gly-Glu-Trp-Gln	Extraction with phosphate buffer followed by hydrolysis with thermolysin and proteinase A, trypsin, proteinase K, tyrosinase, pepsin, papain, protease (sequentially)	MCF-7, AGS, A549	Not mentioned	Inhibition of cell viability	Jang et al., 2008
Bovine lactoferrin	LfcinB FKCRRWQWRMK KLGAPSITCVRRAF	Synthetic	Human umbilical vein endothelial cells	Not stated	Inhibition of angiogenesis	Mader et al, 2006
Milk	Casein phosphopeptide	Chemically synthesized	HT-29 intestinal tumour cells	Not mentioned	Apoptosis	Perego et al, 2011
Bovine milk	Whey protein isolate and collagen hydrolysate	Membrane ultrafiltration and RP-HPLC fractionation	B1F610	0.19–156.9 mg/mL	Apoptosis (increased caspase-3 expression) and necrosis	Castro et al, 2009
Bovine milk	PGPIPN immunomodulatory peptide		SKOV-3	Not stated	Bcl-2 downregulation, apoptosis and inhibition of cell proliferation	Wang et al., 2013

Table 1. Sources, methods of production and mechanisms of action of anticancer food-derived protein hydrolysates and peptides*

Source

Protein hydrolysates/Peptide

Production Method

Cellular and/or animal model

IC₅₀ value

Mechanism

References

*Cell lines Key: MCF-6, MCF-7, MDA-MB-231, BT549–breast cancer; HeLa–cervical cancer; Caco-2, HCT-116, TC7, DLD-1–colon cancer; DU-109–esophageal cancer; NIH3T3–mouse fibroblast; HT1080–fibrosarcoma; U87-glioma; L1210- lymphocytic leukemia; HepG-2–liver cancer; A549, DU-1299–lung cancer; U937–histiocytic lymphoma; HT-29, P388D1-mouse monocyte macrophage cell line; RAW264.7 (mouse)–macrophage; MC3T3E1–osteoblastic calvaria-derived cells; DU-145–prostate; UMR106–rat osteosarcoma-derived cells; AGS–stomach adenocarcinoma; HRA–human ovarian cancer; C33–epidermoid cervical carcinoma; B1F610-melanoma cells; SKVO-3–ovarian cancer cell; HUVEC–Human umbilical vein endothelial cells.

Tuna cooking juice

KPEGMDPPLSEPEDRRDGAAGPK, KLPPLLLAKLLMSGKLLAEPCTGR

Enzymatic hydrolysis with Protease XXIII and ultrafiltration

Human breast cancer MCF-7 cells and MCF-10A mammary epithelial cells

1.39 mg/mL

Apoptosis induction and tumour suppression

Hung et al., 2014

Peptides from Spirulina platensis

HVLSRAPR

Enzymatic hydrolysis and fractionation

HT-29 cancer cells

99.88 µg/mL

Tumour suppression

Wang and Zhang, 2017

Mung bean vicilin

Protein hydrolysates and vicilin seed extracts

Ammonium sulfate fractionation of vicilin followed by separate hydrolyses with alcalase and trypsin

MCF-7 and MDA-MB-231 breast cancer cells

0.32, 0.73 and 0.45 mg/mL for MCF-7; 0.26, 0.48 and 0.54 mg/ml for MDA-MB-231 cells

Tumour suppression

Gupta et al., 2018

Chickpea

Protein hydrolysates

Alcalase hydrolysis

Tumour-bearing mice

Not mentioned

Tumour suppression

Xue et al., 2012

Maize

Peptides

Alcalase hydrolysis followed by ion exchange column chromatography fractionation

Hepatocarcinoma human HepG2 cells

1.5–150 µg/mL for peptide fractions and 0.001–1.5 mM for derived peptides.

Apoptosis induction, cell growth inhibition

Ortiz-Martinez et al., 2017

Soybean (germinated)

Peptide fractions

Extraction, precipitation, enzymatic hydrolysis and membrane fractionation

HeLa cells

1.83 mg/mL

Apoptosis induction and down regulation of PTTG1 and TOP2A tumorigenic genes

Robles-Ramírez et al., 2012

Soybean

Lunasin

Simulated GIT digestion with pepsin and pancreatin

L1210 leukemia cells

14 µM

Activation of Caspase-3 and dose-dependent G2 cell cycle arrest

González de Mejia et al, 2010

Soybean

Lunasin

Lunasin obtained by aqueous extraction from soybean followed by ion exchange and size exclusion chromatography, and further purified by HPLC separation

HT-29 colon cancer cells

61.7 µM

Activation of mitochondrial pathways and increased induction of pro-apoptotic nuclear clusterin expression

Dia and Mejia, 2010

Soybean

XMLPSYSPY

Thermoase and ethanol extraction

P388D1

0.16 mg/mL

Not investigated

Kim et al., 2000

Soybean

Isoflavone-derived soy

Sequential hydrolysis with endopeptidase, exopeptidase and amylase in that order

MCF-7

Not stated

Apoptosis via p21, p53, and caspase-3 expression, and inhibition of VEGF and HSP90

Park et al., 2009

Soybean

Kunitz trypsin inhibitor

HRA human ovarian cancer cells

2–3 µM

Inhibition of the upregulation of urokinase-type plasminogen activator, uPA, and its specific receptor, uPAR

Kobayashi et al, 2004

Soybean

VFDGEL, IETWNPNNKP and FEITPEKNPQ

Pepsin and pancreatin

L1210

7.9, 4.0 and 2.4 mM in that order

Human topoisomerase II inhibition

Wang et al., 2008

Rapeseed

Rapeseed peptide

Alcalase and flavourzyme

HeLa

Not stated

DNA structural damage and apoptosis induction

Xue et al, 2010

Amaranthus mantegazzianus (seed)

Mantegazzianus peptide isolate

Alcalase

UMR106, Caco-2, TC7, MC3T3E1

1.0, 1.5 and 2.5 and 2.5 mg/mL respectively

Inhibition of cell adhesion, necrosis and apoptosis

Barrio and Añón, 2010

Walnut

Cys-Thr-Leu-Glu-Trp

Alkaline extraction, precipitation by centrifugation and hydrolysis with papain

Human HeLa cervical cancer and MCF-7 breast cancer cells

0.60 and 0.65 mg/mL respectively

Induction of apoptosis and autophagy

Ma et al., 2015

Tilapia

Tilapia hepcidin (TH) 1–5

Chemically synthesized

HeLa, HepG2, HT1080

Not stated

Downregulation of interleukin (IL)-6, IL-8, IL-12, IL-15, interferon-γ, Bcl-2 and caspase-7, as well as upregulation of IL-2 and IL-8

Chang et al, 2011

Estuary cod (Epinephelus coioides)

Epinecidin-1

Chemically synthesized

A549, HepG2, HT1080, U937, NIH3T3, RAW264.7 cancer cells

Not stated

Inhibition of cell growth and cell lysis

Lin et al., 2009

Cuttlefish ink (Sepia esculenta)

An oligopeptide, N Gln-Pro-Lys

Trypsin

DU-145 prostate cancer cells

Variable

Inhibition of cell viability and growth

Ding et al, 2011

Tuna dark muscle byproduct

LPHVLTPEAGAT PTAEGGVYMVT

Papain and protease XXIII

MCF-7

8.1 µM and 8.8 µM respectively

Inhibition of cell viability and proliferation

Hsu et al., 2011

Loach (Misgurnus anguillicaudatus)

Loach peptide fractions

Loach tissue homogenization followed by papain hydrolysis

HepG2, MCF-7 and Caco-2 cells

Not stated

Inhibition of cell proliferation

You et al., 2011

Setipinna taty

Heated products from peptic hydrolysates of marine fish (half-fin anchovy)

Peptic hydrolysates heated for 10 min at 95 °C

DU-145 prostate, 1299 lung, and 109 esophagus cancer cells

13.67, 25.17 and 40.28 mg/mL respectively

Cell proliferation inhibition

Song et al., 2011

Skate (Raja porosa) cartilage protein hydrolysate

FIMGPY

Enzymatic hydrolysis with pepsin, trypsin and chymotrypsin

HeLa

4.81 mg/mL

Apoptosis and tumour suppression

Pan et al, 2016

Blue whiting cod, Plaice, Atlantic Salmon

Fish protein hydrolysates

pH-shift extraction followed by Protamex and alcalase hydrolysis

MCF-7/6, MDA-MB-231 and breast cancer cells

Not stated

Inhibition of cell growth and proliferation

Picot et al., 2006

Rice bran

Alacalase hydrolysis followed by simulated GIT digestion

Gastrointestinal juice

Caco-2, HepG2, HCT-116, MCF-7, MDA-MB-231

Not mentioned

Kannan et al., 2010

Snow crab (by-products)

KCl2 cationic peptide

Enzymatic hydrolysis with Protamex, electrodialysis, ultrafiltration

A549, HCT15, BT549, PC3

Not mentioned

Doyen et al., 2011

Squid gelatin

Squid gelatin protein hydrolysates

Protamex, trypsin, Neutrase, Savinase, NS37005, Esperase, Alcalase

MCF-7, U87

130 and 100 µg/mL for MCF-7 and U87 respectively

Not stated

Alemán et al., 2011

Rough sea squirt/solitary tunicate

Solitary tunicate (Styela clava) enzymatic hydrolysates

Alcalase, Thermoase PC10F, pepsin,

AGS, DLD-1, HeLa

577.1–1,240.0 µg/mL

Not mentioned

Kim, 2011

Japanese threadfin bream

Nemipterus japonicus tryptic hydrolysates

4 h hydrolysis with trypsin at 37 °C

HepG2 liver cancer cells

61.1 µg/mL

Inhibition of cell viability

Naqash and Nazeer, 2012

Beef sarcoplasmic protein hydrolysates

Gly-Phe-His-Ile, Asp-Phe-His-Ile-Asn-Gly, Phe-His-Asp, and Gly-Leu-Ser-Asp-Gly-Glu-Trp-Gln

Extraction with phosphate buffer followed by hydrolysis with thermolysin and proteinase A, trypsin, proteinase K, tyrosinase, pepsin, papain, protease (sequentially)

MCF-7, AGS, A549

Not mentioned

Inhibition of cell viability

Jang et al., 2008

Bovine lactoferrin

LfcinB FKCRRWQWRMK KLGAPSITCVRRAF

Synthetic

Human umbilical vein endothelial cells

Not stated

Inhibition of angiogenesis

Mader et al, 2006

Milk

Casein phosphopeptide

Chemically synthesized

HT-29 intestinal tumour cells

Not mentioned

Apoptosis

Perego et al, 2011

Bovine milk

Whey protein isolate and collagen hydrolysate

Membrane ultrafiltration and RP-HPLC fractionation

B1F610

0.19–156.9 mg/mL

Apoptosis (increased caspase-3 expression) and necrosis

Castro et al, 2009

Bovine milk

PGPIPN immunomodulatory peptide

SKOV-3

Not stated

Bcl-2 downregulation, apoptosis and inhibition of cell proliferation

Wang et al., 2013