Expression of Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II oral tongue squamous cell carcinoma

Noboru Habu, Yorihisa Imanishi, Kaori Kameyama, Masayuki Shimoda, Yutaka Tokumaru, Koji Sakamoto, Ryoichi Fujii, Seiji Shigetomi, Kuninori Otsuka, Yoichiro Sato, Yoshihiro Watanabe, Hiroyuki Ozawa, Toshiki Tomita, Masato Fujii, Kaoru Ogawa

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background: The side population (SP) of cancer cells is reportedly enriched with cancer stem cells (CSCs), however, the functional role and clinical relevance of CSC marker molecules upregulated in the SP of head and neck squamous carcinoma (HNSCC) cells are yet to be elucidated. Patients with clinical stage I/II (T1-2N0M0) tongue squamous cell carcinoma (TSCC) typically undergo partial glossectomy; however, development of delayed neck metastasis (DNM) tends to reduce their survival. In the present study, we aimed to determine the CSC markers in the SP of HNSCC cells along with their functions in cellular behaviors, and to clarify the association of these markers with DNM. Methods: Flow cytometry was applied to isolate SP from main population (MP) in HNSCC cells. The expression of the CSC markers was examined by semi-quantitative RT-PCR and immunocytochemistry. In vitro proliferation, migration, and invasion assays were performed to assess cellular behaviors. Clinicopathological factors and immunohistochemical expressions of Oct3/4 and Nanog were evaluated using surgical specimens from 50 patients with stage I/II TSCC. Results: SPs were isolated in all three cell lines examined. Expression levels of Oct3/4 and Nanog were higher in SP cells than MP cells. Additionally, cell migration and invasion abilities were higher in SP cells than MP cells, whereas there was no difference in proliferation. Univariate analysis showed that expression of Oct3/4 and Nanog, vascular and muscular invasion, and mode of invasion were significantly correlated with DNM. Multivariate logistic regression revealed that Oct3/4 expression (risk ratio = 14.78, p = 0.002) and vascular invasion (risk ratio = 12.93, p = 0.017) were independently predictive of DNM. Regarding the diagnostic performance, Oct3/4 showed the highest accuracy, sensitivity, and NPV of 82.0%, 61.5%, and 86.8%, respectively, while vascular invasion showed the highest specificity and PPV of 94.6% and 71.4%, respectively. Conclusion: These results suggest that Oct3/4 and Nanog represent probable CSC markers in HNSCC, which contribute to the development of DNM in part by enhancing cell motility and invasiveness. Moreover, along with vascular invasion, expression of Oct3/4 can be considered a potential predictor for selecting patients at high risk of developing DNM.

Original languageEnglish
Article number730
JournalBMC Cancer
Volume15
Issue number1
DOIs
Publication statusPublished - 2015 Oct 19

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Tongue
Squamous Cell Carcinoma
Neoplastic Stem Cells
Neck
Neoplasm Metastasis
Side-Population Cells
Blood Vessels
Population
Cell Movement
Glossectomy
Odds Ratio
Carcinoma, squamous cell of head and neck
Flow Cytometry
Logistic Models
Immunohistochemistry
Head
Cell Line
Polymerase Chain Reaction
Survival
Neoplasms

Keywords

  • Cancer stem cells
  • Delayed neck metastasis
  • Head and neck squamous cell carcinoma
  • Nanog
  • Oct3/4
  • Side population

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Genetics

Cite this

Expression of Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II oral tongue squamous cell carcinoma. / Habu, Noboru; Imanishi, Yorihisa; Kameyama, Kaori; Shimoda, Masayuki; Tokumaru, Yutaka; Sakamoto, Koji; Fujii, Ryoichi; Shigetomi, Seiji; Otsuka, Kuninori; Sato, Yoichiro; Watanabe, Yoshihiro; Ozawa, Hiroyuki; Tomita, Toshiki; Fujii, Masato; Ogawa, Kaoru.

In: BMC Cancer, Vol. 15, No. 1, 730, 19.10.2015.

Research output: Contribution to journalArticle

Habu, Noboru ; Imanishi, Yorihisa ; Kameyama, Kaori ; Shimoda, Masayuki ; Tokumaru, Yutaka ; Sakamoto, Koji ; Fujii, Ryoichi ; Shigetomi, Seiji ; Otsuka, Kuninori ; Sato, Yoichiro ; Watanabe, Yoshihiro ; Ozawa, Hiroyuki ; Tomita, Toshiki ; Fujii, Masato ; Ogawa, Kaoru. / Expression of Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II oral tongue squamous cell carcinoma. In: BMC Cancer. 2015 ; Vol. 15, No. 1.
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abstract = "Background: The side population (SP) of cancer cells is reportedly enriched with cancer stem cells (CSCs), however, the functional role and clinical relevance of CSC marker molecules upregulated in the SP of head and neck squamous carcinoma (HNSCC) cells are yet to be elucidated. Patients with clinical stage I/II (T1-2N0M0) tongue squamous cell carcinoma (TSCC) typically undergo partial glossectomy; however, development of delayed neck metastasis (DNM) tends to reduce their survival. In the present study, we aimed to determine the CSC markers in the SP of HNSCC cells along with their functions in cellular behaviors, and to clarify the association of these markers with DNM. Methods: Flow cytometry was applied to isolate SP from main population (MP) in HNSCC cells. The expression of the CSC markers was examined by semi-quantitative RT-PCR and immunocytochemistry. In vitro proliferation, migration, and invasion assays were performed to assess cellular behaviors. Clinicopathological factors and immunohistochemical expressions of Oct3/4 and Nanog were evaluated using surgical specimens from 50 patients with stage I/II TSCC. Results: SPs were isolated in all three cell lines examined. Expression levels of Oct3/4 and Nanog were higher in SP cells than MP cells. Additionally, cell migration and invasion abilities were higher in SP cells than MP cells, whereas there was no difference in proliferation. Univariate analysis showed that expression of Oct3/4 and Nanog, vascular and muscular invasion, and mode of invasion were significantly correlated with DNM. Multivariate logistic regression revealed that Oct3/4 expression (risk ratio = 14.78, p = 0.002) and vascular invasion (risk ratio = 12.93, p = 0.017) were independently predictive of DNM. Regarding the diagnostic performance, Oct3/4 showed the highest accuracy, sensitivity, and NPV of 82.0{\%}, 61.5{\%}, and 86.8{\%}, respectively, while vascular invasion showed the highest specificity and PPV of 94.6{\%} and 71.4{\%}, respectively. Conclusion: These results suggest that Oct3/4 and Nanog represent probable CSC markers in HNSCC, which contribute to the development of DNM in part by enhancing cell motility and invasiveness. Moreover, along with vascular invasion, expression of Oct3/4 can be considered a potential predictor for selecting patients at high risk of developing DNM.",
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T1 - Expression of Oct3/4 and Nanog in the head and neck squamous carcinoma cells and its clinical implications for delayed neck metastasis in stage I/II oral tongue squamous cell carcinoma

AU - Habu, Noboru

AU - Imanishi, Yorihisa

AU - Kameyama, Kaori

AU - Shimoda, Masayuki

AU - Tokumaru, Yutaka

AU - Sakamoto, Koji

AU - Fujii, Ryoichi

AU - Shigetomi, Seiji

AU - Otsuka, Kuninori

AU - Sato, Yoichiro

AU - Watanabe, Yoshihiro

AU - Ozawa, Hiroyuki

AU - Tomita, Toshiki

AU - Fujii, Masato

AU - Ogawa, Kaoru

PY - 2015/10/19

Y1 - 2015/10/19

N2 - Background: The side population (SP) of cancer cells is reportedly enriched with cancer stem cells (CSCs), however, the functional role and clinical relevance of CSC marker molecules upregulated in the SP of head and neck squamous carcinoma (HNSCC) cells are yet to be elucidated. Patients with clinical stage I/II (T1-2N0M0) tongue squamous cell carcinoma (TSCC) typically undergo partial glossectomy; however, development of delayed neck metastasis (DNM) tends to reduce their survival. In the present study, we aimed to determine the CSC markers in the SP of HNSCC cells along with their functions in cellular behaviors, and to clarify the association of these markers with DNM. Methods: Flow cytometry was applied to isolate SP from main population (MP) in HNSCC cells. The expression of the CSC markers was examined by semi-quantitative RT-PCR and immunocytochemistry. In vitro proliferation, migration, and invasion assays were performed to assess cellular behaviors. Clinicopathological factors and immunohistochemical expressions of Oct3/4 and Nanog were evaluated using surgical specimens from 50 patients with stage I/II TSCC. Results: SPs were isolated in all three cell lines examined. Expression levels of Oct3/4 and Nanog were higher in SP cells than MP cells. Additionally, cell migration and invasion abilities were higher in SP cells than MP cells, whereas there was no difference in proliferation. Univariate analysis showed that expression of Oct3/4 and Nanog, vascular and muscular invasion, and mode of invasion were significantly correlated with DNM. Multivariate logistic regression revealed that Oct3/4 expression (risk ratio = 14.78, p = 0.002) and vascular invasion (risk ratio = 12.93, p = 0.017) were independently predictive of DNM. Regarding the diagnostic performance, Oct3/4 showed the highest accuracy, sensitivity, and NPV of 82.0%, 61.5%, and 86.8%, respectively, while vascular invasion showed the highest specificity and PPV of 94.6% and 71.4%, respectively. Conclusion: These results suggest that Oct3/4 and Nanog represent probable CSC markers in HNSCC, which contribute to the development of DNM in part by enhancing cell motility and invasiveness. Moreover, along with vascular invasion, expression of Oct3/4 can be considered a potential predictor for selecting patients at high risk of developing DNM.

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KW - Cancer stem cells

KW - Delayed neck metastasis

KW - Head and neck squamous cell carcinoma

KW - Nanog

KW - Oct3/4

KW - Side population

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