Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer

Qin Miao Huang, Shuta Tomida, Yuji Masuda, Chinatsu Arima, Ke Cao, Taka Aki Kasahara, Hirotaka Osada, Yasushi Yatabe, Tomohiro Akashi, Kenji Kamiya, Takashi Takahashi, Motoshi Suzuki

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Genomic instability is an important factor in cancer susceptibility, but a mechanistic understanding of how it arises remains unclear. We examined hypothesized contributions of the replicative DNA polymerase δ (pol δ) subunit POLD4 to the generation of genomic instability in lung cancer. In examinations of 158 lung cancers and 5 mixtures of 10 normal lungs, cell cycle- and checkpoint-related genes generally showed mRNA expression increases in cancer, whereas POLD4 showed reduced mRNA in small cell lung cancer (SCLC). A fraction of non-small cell lung cancer patients also showed low expression comparable with that in SCLC, which was associated with poor prognosis. The lung cancer cell line ACC-LC-48 was found to have low POLD4 expression, with higher histone H3K9 methylation and lower acetylation in the POLD4 promoter, as compared with the A549 cell line with high POLD4 expression. In the absence of POLD4, pol δ exhibited impaired in vitro DNA synthesis activity. Augmenting POLD4 expression in cells where it was attenuated altered the sensitivity to the chemical carcinogen 4-nitroquinoline-1-oxide. Conversely, siRNA-mediated reduction of POLD4 in cells with abundant expression resulted in a cell cycle delay, checkpoint activation, and an elevated frequency of chromosomal gap/break formation. Overexpression of an engineered POLD4 carrying silent mutations at the siRNA target site rescued these phenotypes, firmly establishing the role of POLD4 in these effects. Furthermore, POLD4 overexpression reduced intrinsically high induction of γ-H2AX, a well-accepted marker of double-stranded DNA breaks. Together, our findings suggest that reduced expression of POLD4 plays a role in genomic instability in lung cancer.

Original languageEnglish
Pages (from-to)8407-8416
Number of pages10
JournalCancer Research
Volume70
Issue number21
DOIs
Publication statusPublished - 01-11-2010
Externally publishedYes

Fingerprint

Genomic Instability
DNA-Directed DNA Polymerase
Lung Neoplasms
Small Cell Lung Carcinoma
Small Interfering RNA
4-Nitroquinoline-1-oxide
Chromosome Breakage
cdc Genes
Messenger RNA
Double-Stranded DNA Breaks
Acetylation
Cell Cycle Checkpoints
Non-Small Cell Lung Carcinoma
Carcinogens
Histones
Methylation
Neoplasms
Phenotype
Cell Line
Lung

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Huang, Q. M., Tomida, S., Masuda, Y., Arima, C., Cao, K., Kasahara, T. A., ... Suzuki, M. (2010). Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer. Cancer Research, 70(21), 8407-8416. https://doi.org/10.1158/0008-5472.CAN-10-0784
Huang, Qin Miao ; Tomida, Shuta ; Masuda, Yuji ; Arima, Chinatsu ; Cao, Ke ; Kasahara, Taka Aki ; Osada, Hirotaka ; Yatabe, Yasushi ; Akashi, Tomohiro ; Kamiya, Kenji ; Takahashi, Takashi ; Suzuki, Motoshi. / Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer. In: Cancer Research. 2010 ; Vol. 70, No. 21. pp. 8407-8416.
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Huang, QM, Tomida, S, Masuda, Y, Arima, C, Cao, K, Kasahara, TA, Osada, H, Yatabe, Y, Akashi, T, Kamiya, K, Takahashi, T & Suzuki, M 2010, 'Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer', Cancer Research, vol. 70, no. 21, pp. 8407-8416. https://doi.org/10.1158/0008-5472.CAN-10-0784

Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer. / Huang, Qin Miao; Tomida, Shuta; Masuda, Yuji; Arima, Chinatsu; Cao, Ke; Kasahara, Taka Aki; Osada, Hirotaka; Yatabe, Yasushi; Akashi, Tomohiro; Kamiya, Kenji; Takahashi, Takashi; Suzuki, Motoshi.

In: Cancer Research, Vol. 70, No. 21, 01.11.2010, p. 8407-8416.

Research output: Contribution to journalArticle

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AU - Huang, Qin Miao

AU - Tomida, Shuta

AU - Masuda, Yuji

AU - Arima, Chinatsu

AU - Cao, Ke

AU - Kasahara, Taka Aki

AU - Osada, Hirotaka

AU - Yatabe, Yasushi

AU - Akashi, Tomohiro

AU - Kamiya, Kenji

AU - Takahashi, Takashi

AU - Suzuki, Motoshi

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N2 - Genomic instability is an important factor in cancer susceptibility, but a mechanistic understanding of how it arises remains unclear. We examined hypothesized contributions of the replicative DNA polymerase δ (pol δ) subunit POLD4 to the generation of genomic instability in lung cancer. In examinations of 158 lung cancers and 5 mixtures of 10 normal lungs, cell cycle- and checkpoint-related genes generally showed mRNA expression increases in cancer, whereas POLD4 showed reduced mRNA in small cell lung cancer (SCLC). A fraction of non-small cell lung cancer patients also showed low expression comparable with that in SCLC, which was associated with poor prognosis. The lung cancer cell line ACC-LC-48 was found to have low POLD4 expression, with higher histone H3K9 methylation and lower acetylation in the POLD4 promoter, as compared with the A549 cell line with high POLD4 expression. In the absence of POLD4, pol δ exhibited impaired in vitro DNA synthesis activity. Augmenting POLD4 expression in cells where it was attenuated altered the sensitivity to the chemical carcinogen 4-nitroquinoline-1-oxide. Conversely, siRNA-mediated reduction of POLD4 in cells with abundant expression resulted in a cell cycle delay, checkpoint activation, and an elevated frequency of chromosomal gap/break formation. Overexpression of an engineered POLD4 carrying silent mutations at the siRNA target site rescued these phenotypes, firmly establishing the role of POLD4 in these effects. Furthermore, POLD4 overexpression reduced intrinsically high induction of γ-H2AX, a well-accepted marker of double-stranded DNA breaks. Together, our findings suggest that reduced expression of POLD4 plays a role in genomic instability in lung cancer.

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Huang QM, Tomida S, Masuda Y, Arima C, Cao K, Kasahara TA et al. Regulation of DNA polymerase POLD4 influences genomic instability in lung cancer. Cancer Research. 2010 Nov 1;70(21):8407-8416. https://doi.org/10.1158/0008-5472.CAN-10-0784