MicroRNA-200c attenuates tumour growth and metastasis of presumptive head and neck squamous cell carcinoma stem cells†
Wen-Liang Lo
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan
W-L Lo and C-C Yu contributed equally as first authors.
Search for more papers by this authorCheng-Chia Yu
Institute of Oral Biology and Biomaterial Science, Chung Shan Medical University, Taichung, Taiwan
Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan
W-L Lo and C-C Yu contributed equally as first authors.
Search for more papers by this authorGuang-Yuh Chiou
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorYi-Wei Chen
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorPin-I Huang
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorChian-Shiu Chien
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Search for more papers by this authorLing-Ming Tseng
Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorPen-Yuan Chu
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Laryngology and Head and Neck Surgery, Department of Otolaryngology, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorKai-Hsi Lu
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Medical Research and Education, Cheng-Hsin General Hospital, Taipei, Taiwan
Search for more papers by this authorKuo-Wei Chang
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Search for more papers by this authorShou-Yen Kao
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorCorresponding Author
Shih-Hwa Chiou
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Institute of Pharmacology, National Yang-Ming University and Department of Medical Research and Education, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei 11217, Taiwan.Search for more papers by this authorWen-Liang Lo
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan
W-L Lo and C-C Yu contributed equally as first authors.
Search for more papers by this authorCheng-Chia Yu
Institute of Oral Biology and Biomaterial Science, Chung Shan Medical University, Taichung, Taiwan
Department of Dentistry, Chung Shan Medical University Hospital, Taichung, Taiwan
W-L Lo and C-C Yu contributed equally as first authors.
Search for more papers by this authorGuang-Yuh Chiou
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorYi-Wei Chen
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorPin-I Huang
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorChian-Shiu Chien
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Search for more papers by this authorLing-Ming Tseng
Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorPen-Yuan Chu
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Laryngology and Head and Neck Surgery, Department of Otolaryngology, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorKai-Hsi Lu
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Department of Medical Research and Education, Cheng-Hsin General Hospital, Taipei, Taiwan
Search for more papers by this authorKuo-Wei Chang
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Search for more papers by this authorShou-Yen Kao
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Division of Oral and Maxillofacial Surgery, Department of Stomatology, Taipei Veterans General Hospital, Taipei, Taiwan
Search for more papers by this authorCorresponding Author
Shih-Hwa Chiou
Department of Dentistry and Institute of Oral Biology, School of Dentistry, National Yang-Ming University, Taipei, Taiwan
Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan
Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan
Department of Education and Research, Taipei Veterans General Hospital, Taipei, Taiwan
Institute of Pharmacology, National Yang-Ming University and Department of Medical Research and Education, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei 11217, Taiwan.Search for more papers by this authorNo conflicts of interest were declared.
Abstract
MicroRNA-200c (miR200c) is emerging as an important regulator of tumourigenicity and cancer metastasis with a strong capacity for inducing epithelial–mesenchymal transitions. However, the role of miR200c in head and neck squamous cell carcinoma (HNSCC) and HNSCC-associated cancer stem cells (HNSCC-CSCs) is unknown. In this study, the expression of miR200c in the regional metastatic lymph node of HNSCC tissues was significantly decreased, but BMI1 expression was increased as compared to parental tumours. Importantly, site-directed mutagenesis with a luciferase reporter assay showed that miR200c targeted the 3′ UTR of BMI1 in HNSCC cells. Isolated HNSCC-derived ALDH1+/CD44+ cells displayed CSC-like tumour initiating and radio-resistant properties. The expression levels of miR200c were significantly down-regulated while BMI1 was increased in HNSCC-ALDH1+/CD44+ compared to the other subsets of HNSCC cells. Furthermore, increased miR200c expression or knockdown of BMI1 could significantly inhibit the malignant CSC-like properties of ALDH1+/CD44+ cells. miR200c over-expression further down-regulated the expressions of ZEB1, Snail and N-cadherin, but up-regulated E-cadherin expression in ALDH1+/CD44+ cells. Finally, a xenotransplantion study confirmed that over-expression of miR200c or BMI1 knockdown effectively inhibited the lung metastatic ability and prolonged the survival rate of ALDH1+/CD44+-transplanted mice. In summary, miR200c negatively modulates the expression of BMI1 but also significantly inhibits the metastatic capability of epithelial–mesenchymal transitions in malignant HNSCC by reducing the expression of BMI1/ZEB1. Restoration of miR200c in HNSCC and CSCs may be a promising therapeutic approach. Copyright © 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Supporting Information
| Filename | Description |
|---|---|
| path2826_Suppinfomethods.doc526 KB | Supporting Information: Supplementary materials and methods |
| path2826_Suppinfofs1.tif2.8 MB | Supporting Information: Figure S1. (A) Correlation coefficient values between miR200a, miR200b, or miR200c and Bmi1 in Figure 1A (heatmap of NCI60 database of tumour cells). The data displayed negative correlation between miR200a, miR200b, or miR200c and Bmi1. (B) Correlation coefficient values and p-value between miR200a, miR200b, or miR200c and Bmi1 in Figure 1A was displayed. |
| path2826_Suppinfofs2.tif10.3 MB | Supporting Information: Figure S2. (A) Bmi-1 (upper panel) and miR200c (lower panel) expression levels of 15 independent pairs of normal (N) and cancerous (T) tissues. (B) Transfection efficiency of miR200c and control lentiviral vector by GFP selection marker. |
| path2826_Suppinfofs3.tif3 MB | Supporting Information: Figure S3. (A) Evaluation of the abilities for sphere body formation after serial passage in serum-free medium with bFGF and EGF (patients 3, 4 and 5) by various groups of cells (ALDH1+CD44+, ALDH1+, CD44+, parental, and ALDH1−CD44− cells. (B) Three thousand isolated cells from each cell group, as indicated, were injected into the necks of mice and the tumour volumes were measured over time as indicated (patients 2 and 3). (C) To determine the radiation effect on the tumour growth rate, IR doses of 0–10 Gy were used to treat ALDH1+CD44+, ALDH1+, CD44+, parental or ALDH1−CD44− cells (patients 2 and 3). (D) Detection of BMI1 and ALDH1 co-localization by immunofluorescence. BMI1 protein was prominent and co-existed with ALDH1 protein in the HNSCC cells (upper panel). Average percentage of Bmi1+, ALDH1+, ALDH1+Bmi1+, ALDH1−Bmi1− cells in HNSCC cells (lower panel). |
| path2826_Suppinfofs4.tif1.4 MB | Supporting Information: Figure S4. (A) The linkage distances to ES cells were further expanded with sets of ALDH1+CD44+, ALDH1−CD44−/Bmi-1over, ALDH1+CD44+/miR200cover and ALDH1−CD44− cells. (B) Sox2, Nanog, Oct4, Snail and Nestin expression in control and miR200c-over-expressing ALDH1+CD44+ HNSCCs. |
| path2826_Suppinfots1.doc25 KB | Supporting Information: Table S1. Clinicopathological parameters of 41 HNSCC patients. |
| path2826_Suppinfots2.doc27 KB | Supporting Information: Table S2. The sequences for the primers of quantitative RT–PCR. |
| path2826_Suppinfots3.doc170.5 KB | Supporting Information: Table S3. The expression profile of CD44+ALDH1+ cells. |
| path2826_Suppinfots4.doc232 KB | Supporting Information: Table S4. The expression profile of miR200c-over-expressing CD44+ALDH1+ HNSCC cells. |
Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.
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*Cited in supporting information only.
