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Article

Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling

  • Authors:
    • Akiko Takao
    • Kazuhiro Yoshikawa
    • Sivasundaram Karnan
    • Akinobu Ota
    • Hirotsugu Uemura
    • Marco A. De Velasco
    • Yurie Kura
    • Susumu Suzuki
    • Ryuzo Ueda
    • Tokiko Nishino
    • Yoshitaka Hosokawa
  • View Affiliations / Copyright

    Affiliations: Department of Biochemistry, Aichi Medical University School of Medicine, Nagakute, Aichi 480‑1195, Japan, Division of Research Creation and of Biobank, Research Creation Support Center, Aichi Medical University, Nagakute, Aichi 480‑1195, Japan, Department of Urology, Kindai University Faculty of Medicine, Osaka‑Sayama, Osaka 589‑8511, Japan, Department of Genome Biology, Kindai University Faculty of Medicine, Osaka‑Sayama, Osaka 589‑8511, Japan, Division of Research Support, Research Creation Support Center, Aichi Medical University School of Medicine, Nagakute, Aichi 480‑1195, Japan, Department of Tumor Immunology, Aichi Medical University School of Medicine, Nagakute, Aichi 480‑1195, Japan
    Copyright: © Takao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Pages: 2455-2466
    |
    Published online on: September 5, 2018
       https://doi.org/10.3892/or.2018.6683
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Abstract

Phosphatase and tensin homolog (PTEN) deficiency is associated with development, progression, and metastasis of various cancers. However, changes in gene expression associated with PTEN deficiency have not been fully characterized. To explore genes with altered expression in PTEN‑deficient cells, the present study generated a PTEN‑knockout cell line (ΔPTEN) from a mouse prostate cancer‑derived cell line using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‑associated protein 9 (CRISPR/Cas9) gene editing system. Following transfection of the CRISPR/Cas9 construct, DNA sequencing was performed to identify deletion of the Pten locus and PTEN inactivation was verified by western blotting. The ΔPTEN cell line exhibited enhanced RAC‑alpha serine/threonine‑protein kinase phosphorylation and cyclin D1 expression. In addition, an increase in cell proliferation and colony formation was observed in the ΔPTEN cell line. Gene expression profiling experiments were analyzed with microarray and microRNA (miRNA) arrays. In the microarray analysis, 111 genes exhibited ≥10‑fold increased expression compared with the parent strain and mock cell line and 23 genes were downregulated. The only miRNA with increased expression of 10‑fold or more was mmu‑miR‑210‑3p. Genes with enhanced expression included genes involved in the development, progression, and metastasis of cancer such as Tet methylcytosine dioxygenase 1, twist family BHLH transcription factor 2, C‑fos‑induced growth factor and Wingless‑Type MMTV Integration Site Family, Member 3, and genes involved in immunosuppression such as Arginase 1. The results of the present study suggest that PTEN deficiency mobilizes a variety of genes critical for cancer cell survival and host immune evasion.
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Copy and paste a formatted citation
Spandidos Publications style
Takao A, Yoshikawa K, Karnan S, Ota A, Uemura H, De Velasco MA, Kura Y, Suzuki S, Ueda R, Nishino T, Nishino T, et al: Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling. Oncol Rep 40: 2455-2466, 2018.
APA
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M.A. ... Hosokawa, Y. (2018). Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling. Oncology Reports, 40, 2455-2466. https://doi.org/10.3892/or.2018.6683
MLA
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M. A., Kura, Y., Suzuki, S., Ueda, R., Nishino, T., Hosokawa, Y."Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling". Oncology Reports 40.5 (2018): 2455-2466.
Chicago
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M. A., Kura, Y., Suzuki, S., Ueda, R., Nishino, T., Hosokawa, Y."Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling". Oncology Reports 40, no. 5 (2018): 2455-2466. https://doi.org/10.3892/or.2018.6683
Copy and paste a formatted citation
x
Spandidos Publications style
Takao A, Yoshikawa K, Karnan S, Ota A, Uemura H, De Velasco MA, Kura Y, Suzuki S, Ueda R, Nishino T, Nishino T, et al: Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling. Oncol Rep 40: 2455-2466, 2018.
APA
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M.A. ... Hosokawa, Y. (2018). Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling. Oncology Reports, 40, 2455-2466. https://doi.org/10.3892/or.2018.6683
MLA
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M. A., Kura, Y., Suzuki, S., Ueda, R., Nishino, T., Hosokawa, Y."Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling". Oncology Reports 40.5 (2018): 2455-2466.
Chicago
Takao, A., Yoshikawa, K., Karnan, S., Ota, A., Uemura, H., De Velasco, M. A., Kura, Y., Suzuki, S., Ueda, R., Nishino, T., Hosokawa, Y."Generation of PTEN‑knockout (‑/‑) murine prostate cancer cells using the CRISPR/Cas9 system and comprehensive gene expression profiling". Oncology Reports 40, no. 5 (2018): 2455-2466. https://doi.org/10.3892/or.2018.6683
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