Characterization of human BRCA1 heterozygous knockout cell lines created by somatic cell gene targeting

• Project/Area Number: 22500999
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Carcinogenesis
• Research Institution: Aichi Medical University
• Principal Investigator: KONISHI Hiroyuki 愛知医科大学, 医学部, 准教授 (20344335)
• Co-Investigator(Kenkyū-buntansha): HOSOKAWA Yoshitaka 愛知医科大学, 医学部, 教授 (60229193) ; SIVASUNDARAM Karnan 愛知医科大学, 医学部, 助教 (30557096)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2010: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: BRCA1 / ハプロ不全 / ゲノム不安定性 / ヒト細胞遺伝子改変 / ヒト細胞遺伝子改変法

Research Abstract

To assess the role for heterozygous BRCA1 inactivation in breast carcinogenesis, we created BRCA1 heterozygous knockout clones by targeting the BRCA1 gene in noncanceroushuman breast epithelial cell lines. These clones demonstrated impaired DNA repair via homologous recombination, hypersensitivity to genotoxic agents, and genomic instability, but exhibited no properties of transformed cells. These data suggest that haploinsufficiency caused by BRCA1 heterozygous mutation does not directly lead to cellular transformation or tumor development, but may be involved in the initiation of breast carcinogenesis.

Research Products

• [Journal Article] KRAS 変異がんの治療法 --- 複数の分子標的薬剤を併用する --- (2013)
  • Author(s): 小西裕之
  • Journal Title: 実験医学 Volume: 31 Pages: 538-539
  • URL: https://www.yodosha.co.jp/jikkenigaku/nhpd/9784758100939/c3.html
• [Journal Article] Arsenic trioxide prevents nitric oxide production in LPS-stimulated RAW 264.7 by inhibiting a TRIF-dependent pathway. (2013)
  • Author(s): Takahashi M, Ota A, Karnan S, Ekhtear H, Konishi Y, Damdindorj L, Konishi H, Yokochi T, Nitta M, Hosokawa Y.
  • Journal Title: Cancer Sci Volume: 104 Issue: 2 Pages: 165-70
  • DOI: 10.1111/cas.12053
  • Peer Reviewed
• [Journal Article] KRAS変異がんの治療法 -- 複数の分子標的薬剤を併用する -- (2013)
  • Author(s): 小西裕之
  • Journal Title: 実験医学 Volume: 31 Pages: 538-539
• [Journal Article] Simple monitoring of gene targeting efficiency in human somatic cell lines using the PIGA gene. (2012)
  • Author(s): Karnan S, Konishi Y, Ota A, Takahashi M, Damdindorj L, Hosokawa Y, Konishi H.
  • Journal Title: PLoS ONE Volume: 7 Issue: 10 Pages: e47389-e47389
  • DOI: 10.1371/journal.pone.0047389
  • Peer Reviewed
• [Journal Article] A system for the measurement of gene targeting efficiency in human cell lines using an antibiotic resistance-GFP fusion gene. (2012)
  • Author(s): Konishi Y, Karnan S, Takahashi M, Ota A, Damdindorj L, Hosokawa Y, Konishi H.
  • Journal Title: BioTechniques Volume: 53 Issue: 3 Pages: 141-152
  • DOI: 10.2144/0000113911
  • Peer Reviewed
• [Journal Article] Assessment of the long-term transcriptional activity of a 550-bp-long human ss-actin promoter region. (2012)
  • Author(s): Damdindorj L, Karnan S, Ota A, Takahashi M, Konishi Y, Hossain E, Hosokawa Y, Konishi H.
  • Journal Title: Plasmid Volume: 68 Issue: 3 Pages: 195-200
  • DOI: 10.1016/j.plasmid.2012.07.003
  • Peer Reviewed
• [Journal Article] The growth response to androgen receptor signaling in ERalpha-negative human breast cells is dependent on p21 and mediated by MAPK activation. (2012)
  • Author(s): Garay JP, Karakas B, Abukhdeir AM, Cosgrove DP, Gustin JP, Higgins MJ, Konishi H, Konishi Y, Lauring J, Mohseni M, Wang GM, Jelovac D, Weeraratna A, Sherman Baust CA, Morin PJ, Toubaji A, Meeker A, De Marzo AM, Lewis G, Subhawong A, Argani P, Park BH.
  • Journal Title: Breast Cancer Res Volume: 14 Issue: 1
  • DOI: 10.1186/bcr3112
  • Peer Reviewed
• [Journal Article] Mutation of a single allele of the cancer susceptibility gene BRCA1 leads to genomic instability in human breast epithelial cells. (2011)
  • Author(s): Konishi H, Mohseni M, Tamaki A, Garay JP, Croessmann S, Karnan S, Ota A, Wong HY, Konishi Y, Karakas B, Tahir K, Abukhdeir AM, Gustin JP, Cidado J, Wang GM, Cosgrove D, Cochran R, Jelovac D, Higgins MJ, Arena S, Hawkins L, Lauring J, Gross AL, Heaphy CM, Hosokawa Y, Gabrielson E, Meeker AK, Visvanathan K, Argani P, Bachman KE, Park BH.
  • Journal Title: Proc Natl Acad Sci U S A Volume: 108 Issue: 43 Pages: 17773-8
  • DOI: 10.1073/pnas.1110969108
  • Peer Reviewed
• [Journal Article] PIK3CA mutations and EGFR overexpression predict for lithium sensitivity in human breast epithelial cells. (2011)
  • Author(s): Higgins MJ, Beaver JA, Wong HY, Gustin JP, Lauring JD, Garay JP, Konishi H, Mohseni M, Wang GM, Cidado J, Jelovac D, Cosgrove DP, Tamaki A, Abukhdeir AM, Park BH.
  • Journal Title: Cancer Biol Ther Volume: 11 Issue: 3 Pages: 358-67
  • DOI: 10.4161/cbt.11.3.14227
  • Peer Reviewed
• [Journal Article] PIK3CA mutations and EGFR overexpression predict for lithium sensitivity in human breast epithelial cells (2011)
  • Author(s): Higgins MJ
  • Journal Title: Cancer Biology & Therapy Volume: 11巻 Pages: 358-367
  • Peer Reviewed
• [Journal Article] Novel Metastasis-Related Gene CIM Functions in the Regulation of Multiple Cellular Stress-Response Pathways. (2010)
  • Author(s): Yanagisawa K, Konishi H, Arima C, Tomida S, Takeuchi T, Shimada Y, Yatabe Y, Mitsudomi T, Osada H, Takahashi T.
  • Journal Title: Cancer Res Volume: 70 Issue: 23 Pages: 9949-58
  • DOI: 10.1158/0008-5472.can-10-1055
  • Peer Reviewed
• [Journal Article] Knock in of the AKT1 E17K mutation in human breast epithelial cells does not recapitulate oncogenic PIK3CA mutations. (2010)
  • Author(s): Lauring J, Cosgrove DP, Fontana S, Gustin JP, Konishi H, Abukhdeir AM, Garay JP, Mohseni M, Wang GM, Higgins M, Gorkin D, Reis M, Vogelstein B, Polyak K, Cowherd M, Buckhaults PJ, Park BH.
  • Journal Title: Oncogene Volume: 29 Issue: 16 Pages: 2337-45
  • DOI: 10.1038/onc.2009.516
  • Peer Reviewed
• [Journal Article] Knock in of the AKT1 E17K mutation in human breast epithelial cells does not recapitulate oncogenic PIK3CA mutations (2010)
  • Author(s): Lauring J
  • Journal Title: Oncogene Volume: 29巻 Pages: 2337-2345
  • Peer Reviewed
• [Journal Article] Novel Metastasis-Related Gene CIM Functions in the Regulation of Multiple Cellular Stress-Response Pathways (2010)
  • Author(s): Yanagisawa K
  • Journal Title: Cancer Research Volume: 70巻 Pages: 9949-9958
  • Peer Reviewed
• [Presentation] 三酸化砒素はTRIF依存性経路を抑制しLPS刺激後のRAW264.7細胞からの一酸化窒素の産生を抑制する (2012)
  • Author(s): 高橋美裕希
  • Organizer: 第71回日本癌学会学術総会
  • Place of Presentation: ロイトン札幌（北海道）
• [Presentation] Arsenic exposure accelerates Angiotensin II-mediated Ca2+ signaling in mouse aortic endothelial cells by upregulation of AT1 receptor (2012)
  • Author(s): Hossain E
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] 三酸化ヒ素はSTAT1の活性化を阻害しRAW264.7細胞からの二重鎖RNA刺激によるNO産生を抑制する (2012)
  • Author(s): 太田明伸
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] A comparative investigation of the long-term transcriptional activity of constitutive promoters placed in plasmids and adeno-associated viral vectors (2012)
  • Author(s): Damdindorj L
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] PIGA遺伝子を利用したヒト体細胞株の遺伝子ターゲッティング効率の測定 (2012)
  • Author(s): シバスンダラン・カルナン
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] 抗生剤耐性遺伝子-EGFP融合遺伝子を用いたヒト体細胞株の遺伝子ターゲッティング効率の定量 (2012)
  • Author(s): 小西裕之
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] A mutation in a single BRCA1 allele leads to genomic instability in human breast epithelial cells (2011)
  • Author(s): Konishi H
  • Organizer: 第70回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場(愛知県)
  • Year and Date: 2011-10-04
• [Presentation] CCL8濃度とCCL8陽性細胞の増加はマウス肺のGVHD病態と密接に関与する (2011)
  • Author(s): 太田明伸
  • Organizer: 第70回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場(愛知県)
  • Year and Date: 2011-10-04
• [Presentation] ヒト体細胞株におけるPIGA遺伝子を利用した遺伝子ターゲッティング効率定量系の作成 (2011)
  • Author(s): Sivasundaram Karnan
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都府)
  • Year and Date: 2011-09-24
• [Presentation] BRCA1ヘテロ変異によってヒト乳腺上皮細胞にゲノム不安定性が発生する (2011)
  • Author(s): 小西裕之
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都府)
  • Year and Date: 2011-09-22
• [Presentation] A new protocol of somatic cell gene targeting in human cell lines and its application to breast cancer research (2010)
  • Author(s): Konishi H
  • Organizer: 第15回日韓がんワー クショップ
  • Place of Presentation: 仁川シェラトンホテル(仁川、大韓民国)
  • Year and Date: 2010-12-22
• [Presentation] A new protocol of somatic cell gene targeting in human cell lines and its application to breast cancer research (2010)
  • Author(s): Konishi H
  • Organizer: The 15^<th> Japan-Korea Cancer Research Workshop
  • Place of Presentation: Sheraton Incheon Hotel (Incheon, Korea)
  • Year and Date: 2010-12-22
• [Presentation] ヒト培養細胞遺伝子改変の新しい技術を用いた癌関連遺伝子の機能解析 (2010)
  • Author(s): 小西裕之
  • Organizer: 第33回日本分子生物学会年会第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸国際展示場(兵庫県)
  • Year and Date: 2010-12-07
• [Presentation] ヒト培養細胞遺伝子改変の新しい技術を用いた癌関連遺伝子の機能解析 (2010)
  • Author(s): 小西裕之
  • Organizer: 第83回日本生化学会大会
  • Place of Presentation: 神戸国際展示場(兵庫県)
  • Year and Date: 2010-12-07
• [Presentation] 細胞外ストレス応答を制御する新規肺がん転移関連遺伝子CIMの機能解析 (2010)
  • Author(s): 柳澤聖
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会議場(大阪府)
  • Year and Date: 2010-09-23
• [Presentation] 細胞外ストレス応答を制御する新規肺がん転移関連遺伝子CIMの機能解析 (2010)
  • Author(s): 柳澤聖
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会館(大阪府)
  • Year and Date: 2010-09-23
• [Presentation] ヒト培養細胞遺伝子改変の新技術を用いた癌関連遺伝子の機能解析 (2010)
  • Author(s): 小西裕之
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会議場(大阪府)
  • Year and Date: 2010-09-22
• [Presentation] ヒト培養細胞遺伝子改変の新技術を用いた癌関連遺伝子の機能解析 (2010)
  • Author(s): 小西裕之
  • Organizer: 第69回日本癌学会学術総会
  • Place of Presentation: 大阪国際会館(大阪府)
  • Year and Date: 2010-09-22