Orthodenticle homeobox OTX1 is a potential prognostic biomarker for bladder cancer

ABSTRACT

Bladder cancer (BC) is one of the most aggressive tumors worldwide. OTX1 (orthodenticle homeobox 1) is an important transcription factor involved in various diseases, such as cancers. The aim of this study was to further investigate the role of OTX1 in BC. In this study, differentially expressed genes (DEGs) were screened from tumor tissues and para-cancerous tissues by bioinformatics. The expression of protein and RNA was separately detected by western blotting and immunohistochemistry (IHC), and quantitative polymerase chain reaction (qPCR); cell viability and cell growth were determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and clone formation assays, respectively; cell motility was measured by transwell and wound healing assays; cell cycle was measured by flow cytometry. In this study, 9 DEGs were screened out, and OTX1 was employed as a candidate gene for subsequent study. Results found that OTX1 was highly expressed in BC cells and BC tissues, which was significantly associated with poor prognosis of patients. In addition, OTX1 silencing significantly reduced cell viability, and inhibited cell growth and motility, while OTX1 overexpression got opposite results. Moreover, OTX1 co-expressed genes were enriched in cell cycle-related pathways, suggesting that the role of OTX1 in BC may be related to cell cycle, which was confirmed by flow cytometry analysis. Furthermore, in vivo experiments showed that OTX1 silencing significantly inhibited tumor growth in tumor-bearing mice. Taken together, our findings suggested that OTX1 may play a promotional role in BC progression.

KEYWORDS:

• Bladder cancer
• orthodenticle homeobox 1
• bioinformatics
• cell behavior
• prognostic biomarker

Highlights

1. OTX1 was overexpressed in BC

2. OTX1 was involved in poor prognosis of patients with BC

3. OTX1 promoted the growth and motility of BC cells in vitro

4. OTX1 promoted cell cycle progression of BC cells

5. OTX1 silencing inhibited tumor growth in vivo

Availability of data and materials

All data generated or analyzed during this study are included in this published article.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Ethics approval

All procedures performed in studies involving human participants were in accordance with the standards upheld by the Ethics Committee of the Fourth Affiliated Hospital of Zhejiang University School of Medicine and with those of the 1964 Helsinki Declaration and its later amendments for ethical research involving human subjects.All animal experiments were approved by the Ethics Committee of the Fourth Affiliated Hospital of Zhejiang University School of Medicine for the use of animals and conducted in accordance with the National Institutes of Health Laboratory Animal Care and Use Guidelines.

Statement of informed consent

Written informed consent was obtained from a legally authorized representative(s) for anonymized patient information to be published in this article.

Supplementary material

Supplemental data for this article can be accessed here.

Authors’ contributions

Lei Jiang and Zhongqiang Zuo designed the study, supervised the data collection, Jie Lin analyzed the data, interpreted the data, Chuanfeng Yang prepare the manuscript for publication and reviewed the draft of the manuscript. All authors have read and approved the manuscript.

Additional information

Funding

This work was supported by the Science and Technology program of Jinhua Science and Technology Bureau [Grant No.2020-4-114] and General scientific research Project of Education Department of Zhejiang Province in 2020 [Grant No. Y202045412].