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Hypermethylated ITGA8 Facilitate Bladder Cancer Cell Proliferation and Metastasis

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Abstract

DNA methylation plays a vital role during the development of tumorigenesis. The purpose of this study is to identify candidate DNA methylation drivers during progression of bladder cancer (BLCA). The methylation spectrum in bladder cancer tissues was detected by CHARM analysis, and methylated ITGA8 was selected for further study due to its low expression. Methylation levels in BLCA tissues and cells were detected with methylated-specific PCR (MSP), while mRNA expression and methylation of ITGA8 were detected by qRT-PCR and MSP. After treatment with 5-Aza-dC (DNA methylation inhibitor), the proliferation, migration, and invasion abilities of BLCA cells were determined by MTT, wound healing, and transwell assays, respectively. Flow cytometric analysis was performed to evaluate any variance in the cell cycle. In addition, the effect of demethylated ITGA8 on BLCA tumor growth was verified with an in vivo xenograft tumor model. Based on the methylation profiling of BLCA, ITGA8 was identified to be hypermethylated. ITGA8 methylation levels in BLCA tissues and cells were upregulated, and 5-Aza-dC significantly suppressed ITGA8 methylation levels and increased ITGA8 mRNA expression. Furthermore, after treatment with 5-Aza-dC, the propagation, migration, and invasiveness of the cancer cells were inhibited, and more cancer cells were arrested at the G0/G1 phase. In vivo assays further demonstrated that 5-Aza-dC could impede BLCA tumor growth by repressing methylation levels of ITGA8 and increasing ITGA8 mRNA expression. Hypermethylated ITGA8 facilitated BLCA progression, and 5-Aza-dC treatment inhibited BLCA cell propagation and metastasis by decreasing methylation levels of ITGA8 and inducing cell cycle arrest.

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Data Availability

The datasets used or analysed during the current study are available from the corresponding author on reasonable request.

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Author notes

  1. Xiulong Ma and Liang Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Radiation Oncology, The Second Affiliated Hospital of Xi’an Jiaotong University, Xian, 710004, Shaanxi, China

    Xiulong Ma

  2. Urology Surgery, Jiujiang University Clinic College/Hospital, Jiujiang, 332200, Jiangxi, China

    Liang Zhang

  3. Urology Surgery, Jiujiang University Clinic College/Hospital, Deyang, 618000, Sichuan, China

    Ling Liu

  4. Urology Surgery, Xijing Hospital, Air Force Military Medical University, Xian, 710032, Shaanxi, China

    Dongli Ruan

  5. Urology Surgery, PLA General Hospital, Beijing, 100853, China

    Chunyang Wang

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  1. Xiulong Ma
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Contributions

Contributing to the conception and design: Xiulong Ma; analyzing and interpreting data: Chunyang Wang; drafting the article: Ling Liu, Dongli Ruan; revising it critically for important intellectual content: Liang Zhang; approving the final version to be published: all authors.

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Correspondence to Chunyang Wang.

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This study was authorized by the PLA General Hospital, and obtained written informed consents from all the participants.

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ESM 1

Supplementary Figure S1. The whole uncropped images of the original MS-PCR and qRT-PCR for ITGA8 gene.

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Ma, X., Zhang, L., Liu, L. et al. Hypermethylated ITGA8 Facilitate Bladder Cancer Cell Proliferation and Metastasis. Appl Biochem Biotechnol 196, 245–260 (2024). https://doi.org/10.1007/s12010-023-04512-y

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