An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone

Xiyong Liu, Bingsen Zhou, Shu Mi, Lijun Xue, Jennifer Shih, Janice Lee, Jennifer Chau, Frank Un, Yun Yen

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Mechanistic aberrations leading to Gemcitabine (2′,2′-dFdCyd,2,2-difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance.

Original languageEnglish
Pages (from-to)1927-1938
Number of pages12
JournalBiochemical Pharmacology
Volume73
Issue number12
DOIs
Publication statusPublished - Jun 15 2007
Externally publishedYes

Fingerprint

gemcitabine
Gems
Electron Transport Complex IV
Cytochromes
Sodium Azide
Oxidoreductases
Clone Cells
Adenosine Triphosphate
DNA
KB Cells
Messenger RNA
Hydroxyurea
Azides
Oligonucleotide Array Sequence Analysis
Transcriptome
Respiration
Metabolism
Polymerase Chain Reaction
Genes
Proteins

Keywords

  • ATP
  • Cytochrome C oxidase
  • Drug resistance
  • Gemcitabine
  • Ribonucleotide reductase

ASJC Scopus subject areas

  • Pharmacology

Cite this

An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone. / Liu, Xiyong; Zhou, Bingsen; Mi, Shu; Xue, Lijun; Shih, Jennifer; Lee, Janice; Chau, Jennifer; Un, Frank; Yen, Yun.

In: Biochemical Pharmacology, Vol. 73, No. 12, 15.06.2007, p. 1927-1938.

Research output: Contribution to journalArticle

Liu, Xiyong ; Zhou, Bingsen ; Mi, Shu ; Xue, Lijun ; Shih, Jennifer ; Lee, Janice ; Chau, Jennifer ; Un, Frank ; Yen, Yun. / An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone. In: Biochemical Pharmacology. 2007 ; Vol. 73, No. 12. pp. 1927-1938.
@article{1c38165f84fc4d21835c1e5b20129476,
title = "An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone",
abstract = "Mechanistic aberrations leading to Gemcitabine (2′,2′-dFdCyd,2,2-difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance.",
keywords = "ATP, Cytochrome C oxidase, Drug resistance, Gemcitabine, Ribonucleotide reductase",
author = "Xiyong Liu and Bingsen Zhou and Shu Mi and Lijun Xue and Jennifer Shih and Janice Lee and Jennifer Chau and Frank Un and Yun Yen",
year = "2007",
month = "6",
day = "15",
doi = "10.1016/j.bcp.2007.03.014",
language = "English",
volume = "73",
pages = "1927--1938",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier Inc.",
number = "12",

}

TY - JOUR

T1 - An increase of cytochrome C oxidase mediated disruption of gemcitabine incorporation into DNA in a resistant KB clone

AU - Liu, Xiyong

AU - Zhou, Bingsen

AU - Mi, Shu

AU - Xue, Lijun

AU - Shih, Jennifer

AU - Lee, Janice

AU - Chau, Jennifer

AU - Un, Frank

AU - Yen, Yun

PY - 2007/6/15

Y1 - 2007/6/15

N2 - Mechanistic aberrations leading to Gemcitabine (2′,2′-dFdCyd,2,2-difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance.

AB - Mechanistic aberrations leading to Gemcitabine (2′,2′-dFdCyd,2,2-difluorodeoxycytidine, Gem) resistance may include alteration in its transport, metabolism and incorporation into DNA. To explore the mechanism of Gem resistance, the restriction fragment differential display PCR (RFDD-PCR) was employed to compare the mRNA expression patterns of KBGem (Gem resistant), KBHURs (hydroxyurea resistant) and KBwt (parental KB cell). Nine gene fragments were overexpressed specifically in the KBGem clone. Sequencing and BLAST results showed that three fragments represent cytochrome C oxidase (CCOX, respiration complex IV) subunit III (CCOX3). The cDNA microarray confirmed that the mRNAs of CCOX and ATP synthase subunits were upregulated in KBGem as compared to KBwt and KBHURs. The increase in CCOX1 protein and activity led to the increase of free ATP concentration, which is consistent with the gene expression profile of KBGem. Furthermore, the sensitivity to Gem could be reversed by sodium azide, a CCOX inhibitor. Following the treatment of sodium azide, the cellular accumulation of [3H]-Gem increased in a dose (of azide)-dependent manner, which is associated with increase of [3H]-Gem incorporation into DNA in KBGem. In summary, an increase of CCOX activity and free ATP level may reduce the transport, metabolism and DNA incorporation of Gem, resulting in Gem resistance.

KW - ATP

KW - Cytochrome C oxidase

KW - Drug resistance

KW - Gemcitabine

KW - Ribonucleotide reductase

UR - http://www.scopus.com/inward/record.url?scp=34247567115&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=34247567115&partnerID=8YFLogxK

U2 - 10.1016/j.bcp.2007.03.014

DO - 10.1016/j.bcp.2007.03.014

M3 - Article

VL - 73

SP - 1927

EP - 1938

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

IS - 12

ER -