Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer

Satoshi Yoda, Kenzo Soejima, Junko Hamamoto, Hiroyuki Yasuda, Sohei Nakayama, Ryosuke Satomi, Hideki Terai, Shinnosuke Ikemura, Takashi Sato, Katsuhiko Naoki, Tomoko Betsuyaku

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Objectives: We previously reported low expression of miR-375 in squamous-cell carcinoma (SCC) and high expression in adenocarcinoma (AC) of the lung. miR-. 375's target genes and its function in non-small-cell lung cancer (NSCLC) have not been elucidated. Therefore, the present study was designed to identify the targets of miR-375 and to characterize its function in NSCLC. Materials and methods: Candidate targets of miR-375 were determined using a prediction database and previous data on differential gene expression between SCC and AC. We evaluated miR-375 and target-gene expression levels in 12 NSCLC cell lines. The effect of miR-375 overexpression and knockdown was evaluated in NSCLC cell lines by transfecting them with an miR-375 precursor or inhibitor. A luciferase-reporter assay was performed to confirm a direct interaction between miR-375 and its target gene. Further, a wound-healing assay was performed to evaluate the effect of miR-375 overexpression on the migration of SK-MES-1 cells. Finally, to assess the clinical relevance, 63 clinical NSCLC samples were analyzed. Results: Claudin-1 (CLDN1) has 4 putative miR-375 target sites in its 3'-untranslated region, and this gene was determined to be a target of miR-375. CLDN1 messenger RNA and protein expression were attenuated by overexpression of miR-375 and increased by knockdown of miR-375 in NSCLC cell lines. In a luciferase-reporter assay, miR-375 overexpression resulted in a 3-fold repression of luciferase activity (P<. 0.001). Cell migration was promoted by miR-375 overexpression, suggesting a high potential for invasion and metastasis in NSCLC expressing high levels of miR-375. In clinical NSCLC samples, there was a negative correlation between miR-375 and CLDN1 expression (r = -0.35, P = 0.005). In addition, high miR-375 expression was correlated with a shorter survival time among the clinical samples (P = 0.043). Conclusion: CLDN1 is a novel target of miR-375, and high miR-375 expression shortens survival in NSCLC.

Original languageEnglish
Pages (from-to)366-372
Number of pages7
JournalLung Cancer
Volume85
Issue number3
DOIs
Publication statusPublished - 2014

Fingerprint

Claudin-1
Non-Small Cell Lung Carcinoma
Luciferases
Cell Line
Squamous Cell Carcinoma
Genes
Gene Expression
3' Untranslated Regions
Wound Healing
Cell Movement
Adenocarcinoma
Databases
Neoplasm Metastasis

Keywords

  • Claudin-1
  • Migration
  • MiR-375
  • Non-small-cell lung cancer
  • Survival
  • Target

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine
  • Cancer Research
  • Medicine(all)

Cite this

Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer. / Yoda, Satoshi; Soejima, Kenzo; Hamamoto, Junko; Yasuda, Hiroyuki; Nakayama, Sohei; Satomi, Ryosuke; Terai, Hideki; Ikemura, Shinnosuke; Sato, Takashi; Naoki, Katsuhiko; Betsuyaku, Tomoko.

In: Lung Cancer, Vol. 85, No. 3, 2014, p. 366-372.

Research output: Contribution to journalArticle

Yoda, S, Soejima, K, Hamamoto, J, Yasuda, H, Nakayama, S, Satomi, R, Terai, H, Ikemura, S, Sato, T, Naoki, K & Betsuyaku, T 2014, 'Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer', Lung Cancer, vol. 85, no. 3, pp. 366-372. https://doi.org/10.1016/j.lungcan.2014.06.009
Yoda, Satoshi ; Soejima, Kenzo ; Hamamoto, Junko ; Yasuda, Hiroyuki ; Nakayama, Sohei ; Satomi, Ryosuke ; Terai, Hideki ; Ikemura, Shinnosuke ; Sato, Takashi ; Naoki, Katsuhiko ; Betsuyaku, Tomoko. / Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer. In: Lung Cancer. 2014 ; Vol. 85, No. 3. pp. 366-372.
@article{b1a80c62978647edb04b3b806605f680,
title = "Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer",
abstract = "Objectives: We previously reported low expression of miR-375 in squamous-cell carcinoma (SCC) and high expression in adenocarcinoma (AC) of the lung. miR-. 375's target genes and its function in non-small-cell lung cancer (NSCLC) have not been elucidated. Therefore, the present study was designed to identify the targets of miR-375 and to characterize its function in NSCLC. Materials and methods: Candidate targets of miR-375 were determined using a prediction database and previous data on differential gene expression between SCC and AC. We evaluated miR-375 and target-gene expression levels in 12 NSCLC cell lines. The effect of miR-375 overexpression and knockdown was evaluated in NSCLC cell lines by transfecting them with an miR-375 precursor or inhibitor. A luciferase-reporter assay was performed to confirm a direct interaction between miR-375 and its target gene. Further, a wound-healing assay was performed to evaluate the effect of miR-375 overexpression on the migration of SK-MES-1 cells. Finally, to assess the clinical relevance, 63 clinical NSCLC samples were analyzed. Results: Claudin-1 (CLDN1) has 4 putative miR-375 target sites in its 3'-untranslated region, and this gene was determined to be a target of miR-375. CLDN1 messenger RNA and protein expression were attenuated by overexpression of miR-375 and increased by knockdown of miR-375 in NSCLC cell lines. In a luciferase-reporter assay, miR-375 overexpression resulted in a 3-fold repression of luciferase activity (P<. 0.001). Cell migration was promoted by miR-375 overexpression, suggesting a high potential for invasion and metastasis in NSCLC expressing high levels of miR-375. In clinical NSCLC samples, there was a negative correlation between miR-375 and CLDN1 expression (r = -0.35, P = 0.005). In addition, high miR-375 expression was correlated with a shorter survival time among the clinical samples (P = 0.043). Conclusion: CLDN1 is a novel target of miR-375, and high miR-375 expression shortens survival in NSCLC.",
keywords = "Claudin-1, Migration, MiR-375, Non-small-cell lung cancer, Survival, Target",
author = "Satoshi Yoda and Kenzo Soejima and Junko Hamamoto and Hiroyuki Yasuda and Sohei Nakayama and Ryosuke Satomi and Hideki Terai and Shinnosuke Ikemura and Takashi Sato and Katsuhiko Naoki and Tomoko Betsuyaku",
year = "2014",
doi = "10.1016/j.lungcan.2014.06.009",
language = "English",
volume = "85",
pages = "366--372",
journal = "Lung Cancer",
issn = "0169-5002",
publisher = "Elsevier Ireland Ltd",
number = "3",

}

TY - JOUR

T1 - Claudin-1 is a novel target of miR-375 in non-small-cell lung cancer

AU - Yoda, Satoshi

AU - Soejima, Kenzo

AU - Hamamoto, Junko

AU - Yasuda, Hiroyuki

AU - Nakayama, Sohei

AU - Satomi, Ryosuke

AU - Terai, Hideki

AU - Ikemura, Shinnosuke

AU - Sato, Takashi

AU - Naoki, Katsuhiko

AU - Betsuyaku, Tomoko

PY - 2014

Y1 - 2014

N2 - Objectives: We previously reported low expression of miR-375 in squamous-cell carcinoma (SCC) and high expression in adenocarcinoma (AC) of the lung. miR-. 375's target genes and its function in non-small-cell lung cancer (NSCLC) have not been elucidated. Therefore, the present study was designed to identify the targets of miR-375 and to characterize its function in NSCLC. Materials and methods: Candidate targets of miR-375 were determined using a prediction database and previous data on differential gene expression between SCC and AC. We evaluated miR-375 and target-gene expression levels in 12 NSCLC cell lines. The effect of miR-375 overexpression and knockdown was evaluated in NSCLC cell lines by transfecting them with an miR-375 precursor or inhibitor. A luciferase-reporter assay was performed to confirm a direct interaction between miR-375 and its target gene. Further, a wound-healing assay was performed to evaluate the effect of miR-375 overexpression on the migration of SK-MES-1 cells. Finally, to assess the clinical relevance, 63 clinical NSCLC samples were analyzed. Results: Claudin-1 (CLDN1) has 4 putative miR-375 target sites in its 3'-untranslated region, and this gene was determined to be a target of miR-375. CLDN1 messenger RNA and protein expression were attenuated by overexpression of miR-375 and increased by knockdown of miR-375 in NSCLC cell lines. In a luciferase-reporter assay, miR-375 overexpression resulted in a 3-fold repression of luciferase activity (P<. 0.001). Cell migration was promoted by miR-375 overexpression, suggesting a high potential for invasion and metastasis in NSCLC expressing high levels of miR-375. In clinical NSCLC samples, there was a negative correlation between miR-375 and CLDN1 expression (r = -0.35, P = 0.005). In addition, high miR-375 expression was correlated with a shorter survival time among the clinical samples (P = 0.043). Conclusion: CLDN1 is a novel target of miR-375, and high miR-375 expression shortens survival in NSCLC.

AB - Objectives: We previously reported low expression of miR-375 in squamous-cell carcinoma (SCC) and high expression in adenocarcinoma (AC) of the lung. miR-. 375's target genes and its function in non-small-cell lung cancer (NSCLC) have not been elucidated. Therefore, the present study was designed to identify the targets of miR-375 and to characterize its function in NSCLC. Materials and methods: Candidate targets of miR-375 were determined using a prediction database and previous data on differential gene expression between SCC and AC. We evaluated miR-375 and target-gene expression levels in 12 NSCLC cell lines. The effect of miR-375 overexpression and knockdown was evaluated in NSCLC cell lines by transfecting them with an miR-375 precursor or inhibitor. A luciferase-reporter assay was performed to confirm a direct interaction between miR-375 and its target gene. Further, a wound-healing assay was performed to evaluate the effect of miR-375 overexpression on the migration of SK-MES-1 cells. Finally, to assess the clinical relevance, 63 clinical NSCLC samples were analyzed. Results: Claudin-1 (CLDN1) has 4 putative miR-375 target sites in its 3'-untranslated region, and this gene was determined to be a target of miR-375. CLDN1 messenger RNA and protein expression were attenuated by overexpression of miR-375 and increased by knockdown of miR-375 in NSCLC cell lines. In a luciferase-reporter assay, miR-375 overexpression resulted in a 3-fold repression of luciferase activity (P<. 0.001). Cell migration was promoted by miR-375 overexpression, suggesting a high potential for invasion and metastasis in NSCLC expressing high levels of miR-375. In clinical NSCLC samples, there was a negative correlation between miR-375 and CLDN1 expression (r = -0.35, P = 0.005). In addition, high miR-375 expression was correlated with a shorter survival time among the clinical samples (P = 0.043). Conclusion: CLDN1 is a novel target of miR-375, and high miR-375 expression shortens survival in NSCLC.

KW - Claudin-1

KW - Migration

KW - MiR-375

KW - Non-small-cell lung cancer

KW - Survival

KW - Target

UR - http://www.scopus.com/inward/record.url?scp=84910681176&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84910681176&partnerID=8YFLogxK

U2 - 10.1016/j.lungcan.2014.06.009

DO - 10.1016/j.lungcan.2014.06.009

M3 - Article

VL - 85

SP - 366

EP - 372

JO - Lung Cancer

JF - Lung Cancer

SN - 0169-5002

IS - 3

ER -