Effect of adenoviral N-methylpurine DNA glycosylase overexpression on chemosensitivity of human osteosarcoma cells

Dong Wang, Zhao Yang Zhong, Qin Hong Zhang, Zeng Peng Li, Mark Kelley

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Objective: The overexpression of N-methylpurine DNA glycosylase (MPG) may imbalance the DNA base excision repair (BER) to sensitize tumor cells to current DNA damage chemotherapy. In an effort to improve the efficacy of cancer chemotherapy, we have constructed adenoviral vector of MPG, to study its ability to sensitize human osteosarcoma cell HOS to DNA damage agents. Methods: The adenoviral infection and MPG expression, as well as enzyme activity were determined by flow cytometry, Western blot, and HEX labeled oligonucleotide- based assay respectively. The cell survival/proliferation was measured using MTS, SRB, and [3H] thymidine incorporation assay. Apoptosis cell death was assayed by flow cytometry after treatment using phycoerythin (PE)-conjugated Annexin V and 7-amino-actinomycin (7-AAD). Results: A 10 MOI of recombinant nonreplicating adenovirus was found to infect more than 90% of HOS cells within 24 hours by EGFP fluorescence, in which the MPG overexpression and MPG enzyme activity were also detected. The MPG overexpression HOS cells were significantly more sensitive to the DNA damage agents, including MMS, MNNG, and TMZ, with changes in the IC50 of 6.0, 4.5, and 2.5 fold respectively. Conclusions: These data establish transient MPG overexpression as a potential therapeutic approach for increasing HOS cellular sensitivity to DNA damage agent chemotherapy.

Original languageEnglish (US)
Pages (from-to)352-356
Number of pages5
JournalChinese Journal of Pathology
Volume35
Issue number6
StatePublished - Jun 2006
Externally publishedYes

Fingerprint

DNA Glycosylases
Osteosarcoma
DNA Damage
Drug Therapy
Flow Cytometry
Methylnitronitrosoguanidine
Annexin A5
antineoplaston A10
Dactinomycin
Enzymes
Adenoviridae
Oligonucleotides
DNA Repair
Thymidine
Inhibitory Concentration 50
DNA-3-methyladenine glycosidase II
Neoplasms
Cell Survival
Cell Death
Fluorescence

Keywords

  • Adenoviridae infections
  • Antineoplastic agents
  • Bone neoplasms

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Effect of adenoviral N-methylpurine DNA glycosylase overexpression on chemosensitivity of human osteosarcoma cells. / Wang, Dong; Zhong, Zhao Yang; Zhang, Qin Hong; Li, Zeng Peng; Kelley, Mark.

In: Chinese Journal of Pathology, Vol. 35, No. 6, 06.2006, p. 352-356.

Research output: Contribution to journalArticle

Wang, Dong ; Zhong, Zhao Yang ; Zhang, Qin Hong ; Li, Zeng Peng ; Kelley, Mark. / Effect of adenoviral N-methylpurine DNA glycosylase overexpression on chemosensitivity of human osteosarcoma cells. In: Chinese Journal of Pathology. 2006 ; Vol. 35, No. 6. pp. 352-356.
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