Potent effect of adenoviral vector expressing short hairpin RNA targeting ribonucleotide reductase large subunit M1 on cell viability and chemotherapeutic sensitivity to gemcitabine in non-small cell lung cancer cells

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URI http://shark.lib.kagawa-u.ac.jp/kuir/metadata/28739
Title
Potent effect of adenoviral vector expressing short hairpin RNA targeting ribonucleotide reductase large subunit M1 on cell viability and chemotherapeutic sensitivity to gemcitabine in non-small cell lung cancer cells
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Description

Abstract
BACKGROUND:

Ribonucleotide reductase large subunit (RRM1) is the main enzyme responsible for synthesis of the deoxyribonucleotides used during DNA synthesis. It is also a cellular target for gemcitabine (GEM). Overexpression of RRM1 is reportedly associated with resistance to GEM and the poor prognosis for many types of malignant tumours. Aim of the present study is to establish gene therapy against RRM1-overexpressing tumours.
METHOD:

An adenoviral vector that encoded a short hairpin siRNA targeting the RRM1 gene (Ad-shRRM1) was constructed. Two RRM1-overexpressing non-small cell lung cancer (NSCLC) lines, MAC10 and RERF-LC-MA, were used. Finally, a human tumour xenograft model in nude mice was prepared by subcutaneously implanting tumours derived from RERF-LC-MA cells.
RESULTS:

Ad-shRRM1 effectively downregulated RRM1 mRNA and protein in both types of NSCLC cells and significantly reduced the percentage of viable cells as detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (p<0.005). Caspase 3/7 analysis revealed that transfection with Ad-RRM1 increased the percentage of apoptotic cells in culture containing either type of RRM1-overexpressing cell (p<0.001). Treatment with Ad-shRRM1 exerted a potent antitumour effect against the RRM1-overexpressing RERF-LC-MA xenografts (p<0.05). Furthermore, Ad-shRRM1-mediated inhibition of RRM1 specifically increased sensitivity to gemcitabine of each type of RRM1-overexpressing tumour cell. Combination treatment with Ad-shRRM1 and GEM exerted significantly greater inhibition on cell proliferation than Ad-shRRM1 or GEM treatment alone.
CONCLUSION:

RRM1 appeared to be a promising target for gene therapy, and Ad-shRRM1 had strong antitumour effects, specifically anti-proliferative and pro-apoptotic effects, against NSCLC cells that overexpressed RRM1. Combination therapy with Ad-shRRM1 and GEM may become a new treatment option for patients with NSCLC.

KEYWORDS:

Adenovirus; Chemotherapeutic sensitivity; Gemcitabine; Gene therapy; RRM1; shRNA

(医博甲621)

Author
著者 徳永 義昌
著者(ヨミ) トクナガ ヨシマサ
著者(別表記) Tokunaga Yoshimasa
Publication Title
European Journal of Cancer
Publication Title Alternative
Eur J Cancer.
Volume
51
Issue
6
Start Page
2480
End Page
2489
Publisher
European Association for Cancer Research
Elsevier
Published Date
2015-08-05
ISSN
0959-8049
NCID
AA11526729
PMID
26254808
DOI
10.1016/j.ejca.2015.05.013
Resource Type
Thesis or Dissertation
Language
eng
Resource URL
https://doi.org/10.1016/j.ejca.2015.05.013
Rights
Copyright © 2015 Elsevier Ltd.
この博士論文の本文については、次のエルゼビアの著作権ポリシーの規定により公開しています。「博士論文の場合は、エンバーゴ期間に関係なく機関リポジトリに出版社版を公開することができます。」 The text of this doctoral dissertation is published according to the following Elsevier copyright policy. "In the case of doctoral dissertations, publishers can be published in institutional repositories regardless of the Embargo period."
Publisher's Version with DOI: https://doi.org/10.1016/j.ejca.2015.05.013
Text Version
ETD
Grant ID
博甲第621号
Grant Date
2016-03-24
Degree Name
博士(医学)
Grantor
香川大学
Set
香川大学
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