Korean J Physiol Pharmacol.  2021 Mar;25(2):119-129. 10.4196/kjpp.2021.25.2.119.

Centromere protein U enhances the progression of bladder cancer by promoting mitochondrial ribosomal protein s28 expression

• Liu BB
• Ma T
• Sun W
• Gao WY
• Liu JM
• Li LQ
• Li WY
• Wang S
• Guo YY

• KMID: 2512955
• DOI: http://doi.org/10.4196/kjpp.2021.25.2.119

Abstract

Bladder cancer is one of the most common types of cancer. Most gene mutations related to bladder cancer are dominantly acquired gene mutations and are not inherited. Previous comparative transcriptome analysis of urinary bladder cancer and control samples has revealed a set of genes that may play a role in tumor progression. Here we set out to investigate further the expression of two candidate genes, centromere protein U (CENPU) and mitochondrial ribosomal protein s28 (MRPS28) to better understand their role in bladder cancer pathogenesis. Our results confirmed that CENPU is up-regulated in human bladder cancer tissues at mRNA and protein levels. Gain-of-function and loss-of-function studies in T24 human urinary bladder cancer cell line revealed a hierarchical relationship between CENPU and MRPS28 in the regulation of cell viability, migration and invasion activity. CENPU expression was also up-regulated in in vivo nude mice xenograft model of bladder cancer and mice overexpressing CENPU had significantly higher tumor volume. In summary, our findings identify CENPU and MRPS28 in the molecular pathogenesis of bladder cancer and suggest that CENPU enhances the progression of bladder cancer by promoting MRPS28 expression.

Keyword

Bladder cancer; Cell; Centromere; Protein

Figure

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  Fig. 1 Abnormally high expression of centromere protein U (CENPU) in bladder cancer cells and tissues. Significantly higher CENPU mRNA (A) and protein (B) expression was found in tumor tissues compared to cancer-adjacent normal tissues. Significantly higher CENPU mRNA (C) and protein (D) expression was found in bladder cancer cells (T24) compared to bladder epithelium cells (SV-HUC-1). Scale bar represents 1 cm. **p < 0.01. ***p < 0.001.

• 

  Fig. 2 Overexpression of centromere protein U (CENPU) enhanced the viability, migration, and invasion of T24 cells. (A, B) CENPU mRNA and protein expression of each group was measured by qRT-PCR and Western blotting. (C) The cell viability of each group was measured using the cell counting kit-8 (CCK-8) assay. (D) The cell invasion ability was assessed using Transwell assay, magnification ×100. (E) Cell scratch test was performed to evaluate the cell migration abilities. (F) N-cadherin, Slug, and Snail protein expression in transfected T24 cells of each group determined by Western analysis. *p < 0.05 or **p < 0.01 vs. si-NC group or pcDNA-NC group.

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  Fig. 3 Overexpression of mitochondrial ribosomal protein s28 (MRPS28) promoted the viability, migration, and invasion of T24 cells. (A, B) The protein expression of MRPS28 for each group was determined by Western blot. (C) Cell viability of each group using the cell counting kit-8 (CCK-8) assay. (D) Cell invasion ability using Transwell assay, magnification ×100. (E) Cell scratch test to evaluate the cell migration abilities. *p < 0.05 vs. si-NC group or pcDNA-NC group.

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  Fig. 4 Centromere protein U (CENPU) enhanced the viability, invasion, and migration of T24 cells by promoting mitochondrial ribosomal protein s28 (MRPS28) expression. (A, D) Cell viability of each group was detected using the cell counting kit-8 (CCK-8) assay. (B, E) Cells invasion ability was measured using Transwell assay, magnification ×100. (C, F) Transwell assay was used to evaluate the cell migration abilities, magnification ×100. **p < 0.01 vs. si-NC group or pcDNA-NC group. ##p < 0.01 vs. si-CENPU group or pcDNA-CENPU group.

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  Fig. 5 Centromere protein U (CENPU) promoted bladder cancer development in vivo by elevating mitochondrial ribosomal protein s28 (MRPS28) expression. (A) Picture of tumor tissues and volume of tumors from nude mice in each group from 1–4 weeks. (B, C) Immunohistochemistry and Western blot analysis was used to measure the expression of CENPU and MRPS28 proteins in tumor tissues of nude mice. (D) Western blot analysis to measure N-cadherin, Slug and Snail protein expression in tumor tissues. In this study, T24 cells were injected subcutaneously into the back of nude mice. *p < 0.05 or ***p < 0.001 vs. pcDNA-NC group. #p < 0.05 or ##p < 0.01 or ###p < 0.001 vs. pcDNA-CENPU group.

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