Biliverdin Reductase A (BLVRA) Promotes Colorectal Cancer Cell Progression by Activating the Wnt/β-Catenin Signaling Pathway

Haiyan Mao; Yuan Xu; Zhengrong Zhang; Guozhuang Sun; Zhu Wang; Dawei Qiao; Xudong Yin; Siping Liu; Ping Bo

doi:10.2147/CMAR.S242531

Biliverdin Reductase A (BLVRA) Promotes Colorectal Cancer Cell Progression by Activating the Wnt/β-Catenin Signaling Pathway

Cancer Manag Res. 2020 Apr 23:12:2697-2709. doi: 10.2147/CMAR.S242531. eCollection 2020.

Authors

Haiyan Mao^{1

2}, Yuan Xu¹, Zhengrong Zhang¹, Guozhuang Sun³, Zhu Wang¹, Dawei Qiao², Xudong Yin¹, Siping Liu⁴, Ping Bo²

Affiliations

¹ Oncology Department, The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou, Jiangsu Province 225000, People's Republic of China.
² Department of Integrative Chinese and Western Medicine, Medical College of Yangzhou University, Yangzhou, Jiangsu Province 225009, People's Republic of China.
³ Laboratory Department, Xuyi People's Hospital, Huai'an, Jiangsu Province 211700, People's Republic of China.
⁴ Imaging Department, Yangzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine, Yangzhou 225000, People's Republic of China.

Abstract

Purpose: Biliverdin reductase A (BLVRA) is a pleiotropic enzyme that converts biliverdin-IX-alpha into the antioxidant and anti-nitrosative compound, bilirubin-IX-alpha. It is related to various diseases, including cancer. It is overexpressed in many types of cancers and promotes cancer development and metastasis, but the effects of BLVRA in colorectal cancer have not been researched at present. This study was aimed to investigate the effects of biliverdin reductase A (BLVRA) in vivo and vitro experiments and its possible mechanism.

Methods: The clinical samples of CRC patients and CRC cell lines HT-29 and SW620 were chosen to perform the experiments. ELISA and Immunohistochemistry (IHC) were applied to test the level of BLVRA in patients. HT-29 knockdown of BLVRA and SW620 overexpression of BLVRA was established by the lentiviral vector transfection. Reverse transcription-quantitative real-time polymerase chain reaction and Western blotting were performed to examine the expression of BLVRA. MTT was used to detect the proliferation of CRC cells. Flow cytometry was applied to assess the rate of apoptosis. Transwell assay was performed to examine the capacity of migration and invasion. Immunofluorescence staining was adopted to assess the expression of E-cadherin and vimentin. Western blotting was utilized to detect the expression of apoptosis-related proteins, EMT-related proteins and target proteins of Wnt/β-catenin signaling pathway.

Results: Analysis of the clinical samples revealed that BLVRA was overexpressed in CRC patients and implied poor prognosis. BLVRA overexpression in the in vitro studies revealed that it increased the potential of CRC cells for proliferation, migration and invasion; augmented EMT; and hindered apoptosis. In addition, BLVRA overexpression was found to upregulate positive target genes and downregulate negative target genes of the Wnt/β-catenin signaling pathway, which implied that the biological effects of BLVRA in CRC were mediated by this pathway. In contrast, knockdown of BLVRA manifested the opposite effects.

Conclusion: Our results suggested that BLVRA might be a promising prognostic marker and a potential therapeutic target in CRC.

Keywords: Wnt/β-catenin signaling pathway; biliverdin reductase A; cancer progression; colorectal cancer.