Structurally related antitumor effects of flavanones in vitro and in vivo: Involvement of caspase 3 activation, p21 gene expression, and reactive oxygen species production

Shing Chuan Shen, Ching Huai Ko, Shi Wen Tseng, Shu Huei Tsai, Yen Chou Chen

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Flavonoids exist extensively in plants and Chinese herbs, and several biological effects of flavonoids have been demonstrated. The antitumor effects in colorectal carcinoma cells (HT29, COLO205, and COLO320HSR) of eight flavanones including flavanone, 2′-OH flavanone, 4′-OH flavanone, 6-OH flavanone, 7-OH flavanone, naringenin, nargin, and taxifolin were investigated. Results of the MTT assay indicate that 2′-OH flavanone showed the most potent cytotoxic effect on these three cells, and cell death induced by 2′-OH flavanone was via the occurrence of DNA ladders, apoptotic bodies, and hypodiploid cells, all characteristics of apoptosis. Induction of caspase 3 protein processing and enzyme activity associated with cleavage of poly(ADP-ribose) polymerase (PARP) was identified in 2′-OH flavanone-treated cells, and a peptidyl inhibitor (Ac-DEVD-FMK) of caspase 3 attenuated the cytotoxicity of 2′-OH flavanone in COLO205 and HT-29 cells. Elevation of p21 (but not p53) and a decrease in Mcl-1 protein were found in 2′-OH flavanone-treated COLO205 and HT-29 cells. Elevation of intracellular reactive oxygen species (ROS) was detected in 2′-OH flavanone-treated cells by the 2′,7′-dichlorodihydrofluorescein diacetate (DCHF-DA) assay, and ROS scavengers including 4,5-dihydro-1,3-benzene disulfonic acid (tiron), catalase, superoxide dismutase (SOD), and pyrrolidine dithiocarbamate (PDTC) suppressed the 2′-OH flavanone-induced cytotoxic effect. Subcutaneous injection of COLO205 induced tumor formation in nude mice, and 2′-OH flavanone showed a significant inhibitory effect on tumor formation. The appearance of apoptotic cells with H&E staining, and an increase in p21, but not p53, protein by immunohistochemistry were observed in tumor tissues under 2′-OH flavanone treatment. Primary tumor cells (COLO205-X) derived from a tumor specimen elicited by COLO205 were established, and 2′-OH flavanone showed an significant apoptotic effect in COLO205-X cells in accordance with the appearance of DNA ladders, caspase 3 protein processing, PARP protein cleavage, and increasing p21 protein. These results revealed in vitro, ex vivo, and in vivo antitumor activities of 2′-OH flavanone via apoptosis induction, and indicates that 2′-OH flavanone is an active compound worthy of development for cancer chemotherapy.

Original languageEnglish
Pages (from-to)84-95
Number of pages12
JournalToxicology and Applied Pharmacology
Volume197
Issue number2
DOIs
Publication statusPublished - Jun 1 2004

Fingerprint

Flavanones
Gene expression
Caspase 3
Reactive Oxygen Species
Chemical activation
Gene Expression
Tumors
HT29 Cells
Cells
Neoplasms
Proteins
Poly(ADP-ribose) Polymerases
Ladders
Flavonoids
2'-hydroxyflavanone
In Vitro Techniques
Assays
1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium Salt
Apoptosis
Chemotherapy

Keywords

  • 2′-OH flavanone
  • Apoptosis
  • Caspase 3
  • COX-1
  • In vitro
  • In vivo
  • nonsteroid anti-inflammatory drugs
  • NSAIDs
  • p21
  • PARP
  • PGE
  • poly(ADP-ribose) polymerase
  • prostaglandin E
  • reactive oxygen species
  • ROS
  • SOD
  • superoxide dismutase

ASJC Scopus subject areas

  • Pharmacology
  • Toxicology

Cite this

@article{896622c375414c11b0a00b1420e02616,
title = "Structurally related antitumor effects of flavanones in vitro and in vivo: Involvement of caspase 3 activation, p21 gene expression, and reactive oxygen species production",
abstract = "Flavonoids exist extensively in plants and Chinese herbs, and several biological effects of flavonoids have been demonstrated. The antitumor effects in colorectal carcinoma cells (HT29, COLO205, and COLO320HSR) of eight flavanones including flavanone, 2′-OH flavanone, 4′-OH flavanone, 6-OH flavanone, 7-OH flavanone, naringenin, nargin, and taxifolin were investigated. Results of the MTT assay indicate that 2′-OH flavanone showed the most potent cytotoxic effect on these three cells, and cell death induced by 2′-OH flavanone was via the occurrence of DNA ladders, apoptotic bodies, and hypodiploid cells, all characteristics of apoptosis. Induction of caspase 3 protein processing and enzyme activity associated with cleavage of poly(ADP-ribose) polymerase (PARP) was identified in 2′-OH flavanone-treated cells, and a peptidyl inhibitor (Ac-DEVD-FMK) of caspase 3 attenuated the cytotoxicity of 2′-OH flavanone in COLO205 and HT-29 cells. Elevation of p21 (but not p53) and a decrease in Mcl-1 protein were found in 2′-OH flavanone-treated COLO205 and HT-29 cells. Elevation of intracellular reactive oxygen species (ROS) was detected in 2′-OH flavanone-treated cells by the 2′,7′-dichlorodihydrofluorescein diacetate (DCHF-DA) assay, and ROS scavengers including 4,5-dihydro-1,3-benzene disulfonic acid (tiron), catalase, superoxide dismutase (SOD), and pyrrolidine dithiocarbamate (PDTC) suppressed the 2′-OH flavanone-induced cytotoxic effect. Subcutaneous injection of COLO205 induced tumor formation in nude mice, and 2′-OH flavanone showed a significant inhibitory effect on tumor formation. The appearance of apoptotic cells with H&E staining, and an increase in p21, but not p53, protein by immunohistochemistry were observed in tumor tissues under 2′-OH flavanone treatment. Primary tumor cells (COLO205-X) derived from a tumor specimen elicited by COLO205 were established, and 2′-OH flavanone showed an significant apoptotic effect in COLO205-X cells in accordance with the appearance of DNA ladders, caspase 3 protein processing, PARP protein cleavage, and increasing p21 protein. These results revealed in vitro, ex vivo, and in vivo antitumor activities of 2′-OH flavanone via apoptosis induction, and indicates that 2′-OH flavanone is an active compound worthy of development for cancer chemotherapy.",
keywords = "2′-OH flavanone, Apoptosis, Caspase 3, COX-1, In vitro, In vivo, nonsteroid anti-inflammatory drugs, NSAIDs, p21, PARP, PGE, poly(ADP-ribose) polymerase, prostaglandin E, reactive oxygen species, ROS, SOD, superoxide dismutase",
author = "Shen, {Shing Chuan} and Ko, {Ching Huai} and Tseng, {Shi Wen} and Tsai, {Shu Huei} and Chen, {Yen Chou}",
year = "2004",
month = "6",
day = "1",
doi = "10.1016/j.taap.2004.02.002",
language = "English",
volume = "197",
pages = "84--95",
journal = "Toxicology and Applied Pharmacology",
issn = "0041-008X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Structurally related antitumor effects of flavanones in vitro and in vivo

T2 - Involvement of caspase 3 activation, p21 gene expression, and reactive oxygen species production

AU - Shen, Shing Chuan

AU - Ko, Ching Huai

AU - Tseng, Shi Wen

AU - Tsai, Shu Huei

AU - Chen, Yen Chou

PY - 2004/6/1

Y1 - 2004/6/1

N2 - Flavonoids exist extensively in plants and Chinese herbs, and several biological effects of flavonoids have been demonstrated. The antitumor effects in colorectal carcinoma cells (HT29, COLO205, and COLO320HSR) of eight flavanones including flavanone, 2′-OH flavanone, 4′-OH flavanone, 6-OH flavanone, 7-OH flavanone, naringenin, nargin, and taxifolin were investigated. Results of the MTT assay indicate that 2′-OH flavanone showed the most potent cytotoxic effect on these three cells, and cell death induced by 2′-OH flavanone was via the occurrence of DNA ladders, apoptotic bodies, and hypodiploid cells, all characteristics of apoptosis. Induction of caspase 3 protein processing and enzyme activity associated with cleavage of poly(ADP-ribose) polymerase (PARP) was identified in 2′-OH flavanone-treated cells, and a peptidyl inhibitor (Ac-DEVD-FMK) of caspase 3 attenuated the cytotoxicity of 2′-OH flavanone in COLO205 and HT-29 cells. Elevation of p21 (but not p53) and a decrease in Mcl-1 protein were found in 2′-OH flavanone-treated COLO205 and HT-29 cells. Elevation of intracellular reactive oxygen species (ROS) was detected in 2′-OH flavanone-treated cells by the 2′,7′-dichlorodihydrofluorescein diacetate (DCHF-DA) assay, and ROS scavengers including 4,5-dihydro-1,3-benzene disulfonic acid (tiron), catalase, superoxide dismutase (SOD), and pyrrolidine dithiocarbamate (PDTC) suppressed the 2′-OH flavanone-induced cytotoxic effect. Subcutaneous injection of COLO205 induced tumor formation in nude mice, and 2′-OH flavanone showed a significant inhibitory effect on tumor formation. The appearance of apoptotic cells with H&E staining, and an increase in p21, but not p53, protein by immunohistochemistry were observed in tumor tissues under 2′-OH flavanone treatment. Primary tumor cells (COLO205-X) derived from a tumor specimen elicited by COLO205 were established, and 2′-OH flavanone showed an significant apoptotic effect in COLO205-X cells in accordance with the appearance of DNA ladders, caspase 3 protein processing, PARP protein cleavage, and increasing p21 protein. These results revealed in vitro, ex vivo, and in vivo antitumor activities of 2′-OH flavanone via apoptosis induction, and indicates that 2′-OH flavanone is an active compound worthy of development for cancer chemotherapy.

AB - Flavonoids exist extensively in plants and Chinese herbs, and several biological effects of flavonoids have been demonstrated. The antitumor effects in colorectal carcinoma cells (HT29, COLO205, and COLO320HSR) of eight flavanones including flavanone, 2′-OH flavanone, 4′-OH flavanone, 6-OH flavanone, 7-OH flavanone, naringenin, nargin, and taxifolin were investigated. Results of the MTT assay indicate that 2′-OH flavanone showed the most potent cytotoxic effect on these three cells, and cell death induced by 2′-OH flavanone was via the occurrence of DNA ladders, apoptotic bodies, and hypodiploid cells, all characteristics of apoptosis. Induction of caspase 3 protein processing and enzyme activity associated with cleavage of poly(ADP-ribose) polymerase (PARP) was identified in 2′-OH flavanone-treated cells, and a peptidyl inhibitor (Ac-DEVD-FMK) of caspase 3 attenuated the cytotoxicity of 2′-OH flavanone in COLO205 and HT-29 cells. Elevation of p21 (but not p53) and a decrease in Mcl-1 protein were found in 2′-OH flavanone-treated COLO205 and HT-29 cells. Elevation of intracellular reactive oxygen species (ROS) was detected in 2′-OH flavanone-treated cells by the 2′,7′-dichlorodihydrofluorescein diacetate (DCHF-DA) assay, and ROS scavengers including 4,5-dihydro-1,3-benzene disulfonic acid (tiron), catalase, superoxide dismutase (SOD), and pyrrolidine dithiocarbamate (PDTC) suppressed the 2′-OH flavanone-induced cytotoxic effect. Subcutaneous injection of COLO205 induced tumor formation in nude mice, and 2′-OH flavanone showed a significant inhibitory effect on tumor formation. The appearance of apoptotic cells with H&E staining, and an increase in p21, but not p53, protein by immunohistochemistry were observed in tumor tissues under 2′-OH flavanone treatment. Primary tumor cells (COLO205-X) derived from a tumor specimen elicited by COLO205 were established, and 2′-OH flavanone showed an significant apoptotic effect in COLO205-X cells in accordance with the appearance of DNA ladders, caspase 3 protein processing, PARP protein cleavage, and increasing p21 protein. These results revealed in vitro, ex vivo, and in vivo antitumor activities of 2′-OH flavanone via apoptosis induction, and indicates that 2′-OH flavanone is an active compound worthy of development for cancer chemotherapy.

KW - 2′-OH flavanone

KW - Apoptosis

KW - Caspase 3

KW - COX-1

KW - In vitro

KW - In vivo

KW - nonsteroid anti-inflammatory drugs

KW - NSAIDs

KW - p21

KW - PARP

KW - PGE

KW - poly(ADP-ribose) polymerase

KW - prostaglandin E

KW - reactive oxygen species

KW - ROS

KW - SOD

KW - superoxide dismutase

UR - http://www.scopus.com/inward/record.url?scp=2442507762&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442507762&partnerID=8YFLogxK

U2 - 10.1016/j.taap.2004.02.002

DO - 10.1016/j.taap.2004.02.002

M3 - Article

C2 - 15163544

AN - SCOPUS:2442507762

VL - 197

SP - 84

EP - 95

JO - Toxicology and Applied Pharmacology

JF - Toxicology and Applied Pharmacology

SN - 0041-008X

IS - 2

ER -