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Selective cell death of p53-insufficient cancer cells is induced by knockdown of the mRNA export molecule GANP

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Abstract

Cancer cells often contain p53 abnormalities that impair cell-cycle checkpoint progression and cause resistance to various anti-cancer treatments. DNA damage occurs at actively transcribed genes during G1-phase in yeast cells that have a deficient mRNA export capacity. Here, we show that germinal center-associated nuclear protein (GANP), a homologue of yeast Sac3 that is involved in mRNA export, is indispensable for ensuring the stability of human genomic DNA and that GANP knockdown causes apoptosis and necrosis of p53-insufficient cancer cells. Ganp small interfering RNA (siGanp)-induced DNA damage, accompanied by a decrease in the number of cells in S-phase, caused late apoptosis and necrosis in p53-insufficient cancer cells through both caspase-dependent and -independent mechanisms. siGanp effectively induced DNA damage leading to cell death in p53-insufficient cancer cells in vitro and protect the growth of cancer cells transplanted into immunocompromized mice, suggesting that siGanp has potential as a selective treatment for p53-insufficient cancer cells.

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Abbreviations

Ab:

Antibody

AID:

Activation-induced cytidine deaminase

BrdU:

Bromodeoxyuridine

CCA:

Cholangiocarcinoma

CPT:

Camptothecin

C-terminal:

Carboxyl-terminal

DSB:

Double-strand break

GANP:

Germinal center-associated nuclear protein

N-terminal:

Amino-terminal

PI:

Propidium iodide

siRNA:

Small interfering RNA

shRNA:

Short hairpin RNA

Wt:

Wild-type

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Acknowledgments

We would like to thank Mika Ito and Yoshimi Fukushima for technical assistance. This work was supported by a Grant-in-Aid for Science Research in a Priority Area (Immunology Community) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) (to N.S.), Advanced Education Program for Integrated Clinical, Basic and Social Medicine, Graduate School of Medical Sciences, Kumamoto University (Program for Enhancing Systematic Education in Graduate Schools, MEXT) (to N.S.), the Adaptable and Seamless Technology Transfer Program through Target-driven R&D (Japan Science and Technology Agency) (to K.K.), Heiwa Nakajima Foundation (to S.O., K.V., and K.K.), a contract research fund from the Program of Founding Research Centers for Emerging and Reemerging Infectious Diseases (to N.S.), and a Global COE program (Global Education and Research Center Aiming at the control of AIDS in Kumamoto University) (to N.S.). Suchada Phimsen is supported by a scholarship for The International Priority Graduate Programs (PGP); Advanced Graduate Courses for International Students (Doctoral Course), MEXT, Japan.

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Authors and Affiliations

  1. Department of Immunology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1, Honjo, Kumamoto, 860-8556, Japan

    Suchada Phimsen, Kazuhiko Kuwahara, Teruo Nakaya, Kazutaka Ohta, Andri Rezano, Masahiro Kitabatake & Nobuo Sakaguchi

  2. Department of Biochemistry and Electron Microscope Center, Kawasaki Medical School, 577 Matsushima, Kurashiki, Okayama, 701-0192, Japan

    Taiji Suda & Shigenobu Tone

  3. Division of Hematopoiesis, Center for AIDS Research, Kumamoto University, 2-2-1, Honjo, Kumamoto, 860-0811, Japan

    Kulthida Vaeteewoottacharn & Seiji Okada

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  1. Suchada Phimsen
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Correspondence to Nobuo Sakaguchi.

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Phimsen, S., Kuwahara, K., Nakaya, T. et al. Selective cell death of p53-insufficient cancer cells is induced by knockdown of the mRNA export molecule GANP. Apoptosis 17, 679–690 (2012). https://doi.org/10.1007/s10495-012-0711-8

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