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WDR12/RAC1 axis promoted proliferation and anti-apoptosis in colorectal cancer cells

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Abstract

Background: WD repeat domain 12 (WDR12) plays a crucial role in the ribosome biogenesis pathway. However, its biological function in colorectal cancer (CRC) remains poorly understood. Therefore, this study aims to investigate the roles of WDR12 in the occurrence and progression of CRC, as well as its underlying mechanisms. Methods: The expression of WDR12 was assessed through The Cancer Genome Atlas (TCGA) and the Human Protein Atlas (HPA) database. Functional experiments including Celigo assay, MTT assay, and Caspase-3/7 assay were conducted to validate the role of WDR12 in the malignant progression of CRC. Additionally, mRNA chip-sequencing and ingenuity pathway analysis (IPA) were performed to identify the molecular mechanism. Results: WDR12 expression was significantly upregulated in CRC tissues compared to normal colorectal tissues. Knockdown of WDR12 reduced proliferation and promoted apoptosis of CRC cell lines in vitro and in vivo experiments. Furthermore, WDR12 expression had a significantly inverse association with diseases and functions, including cancer, cell cycle, DNA replication, recombination, cellular growth, proliferation and repair, as revealed by IPA analysis of mRNA chip-sequencing data. Moreover, the activation of cell cycle checkpoint kinases proteins in the cell cycle checkpoint control signaling pathway was enriched in the WDR12 knockdown CRC cell lines. Additionally, downregulation of rac family small GTPase 1 (RAC1) occurred upon WDR12 knockdown, thereby facilitating the proliferation and anti-apoptosis of CRC cells. Conclusion: Our study demonstrates that the WDR12/RAC1 axis promotes tumor progression in CRC. Therefore, WDR12 may serve as a novel oncogene and a potential target for individualized therapy in CRC. These findings provide an experimental foundation for the clinical development of drugs targeting the WDR12/RAC1 axis.

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Data availability

Data available in a publicly accessible repository (TCGA database; https://portal.gdc.cancer.gov/).

Abbreviations

CDKN2A:

Cyclin dependent kinase inhibitor 2A

CRC:

Colorectal cancer

DEGs:

Differentially expressed genes

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HPA:

The human protein atlas

IPA:

Ingenuity pathway analysis

mRNA:

Messenger ribonucleic acid

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide

PCR:

Polymerase chain reaction

PLK1:

Polo like kinase 1

PPI:

Protein–protein interaction networks

PVDF:

Polyvinylidene fluoride

Rho:

Rhodopsin

RAC1:

Rac family small GTPase 1

RT-qPCR:

Real time quantitative polymerase chain reaction

shCtrl:

Empty lentivirus transfection control

shWDR12:

Knockdown WDR12

siRNA:

Small interfering RNA

TCGA:

The cancer genome atlas

TISCH:

The single cell database

UALCAN:

The University of Alabama at Birmingham cancer data analysis portal

WDR12:

WD repeat domain 12

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Acknowledgements

We acknowledge the technical support from Shanghai Genechem Co.,Ltd and the contribution from TCGA database.

Funding

This research was funded by the Chen Xiao-ping Foundation for the Development of Science and Technology of Hubei Province, grant number CXPJJH122006-1001.

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Authors and Affiliations

  1. Department of Geriatrics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Jiefang Road No.1095, Wuhan, 430030, Hubei, China

    Su Wen, Xueqing Huang, Liping Xiong, Hao Zeng, Shuang Wu, Kangli An & Tiejun Yin

  2. Geneplus-Beijing Institute, Zhongguancun Life Science Park, Peking University Medical Industrial Park, Life Park Road No.8, Beijing, 102205, China

    Jing Bai & Zhipeng Zhou

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  1. Su Wen
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Contributions

TY planned the experiments and revised the manuscript. SW (Su Wen) and XH conducted the experiments. SW (Su Wen) wrote the original manuscript. SW (Su Wen), XH, LX and HZ performed an extensive literature review. KA and SW (Shuang Wu) conducted statistical analysis. JB, ZZ, XX reviewed the manuscript, revising it for important intellectual content. All authors read and approved the final manuscript.

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Correspondence to Tiejun Yin.

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The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Review Board of Shanghai Genechem Co., Ltd (protocol code GSZE0167660 and date of July 8, 2018).

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Wen, S., Huang, X., Xiong, L. et al. WDR12/RAC1 axis promoted proliferation and anti-apoptosis in colorectal cancer cells. Mol Cell Biochem (2024). https://doi.org/10.1007/s11010-024-04937-x

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