RhoB expression associated with chemotherapy response and prognosis in colorectal cancer

Maria Kopsida; Na Liu; Angeliki Kotti; Jing Wang; Lasse Jensen; Ganesan Jothimani; Camilla Hildesjo; Staffan Haapaniemi; Wen Zhong; Surajit Pathak; Xiao-Feng Sun

doi:10.1186/s12935-024-03236-1

RhoB expression associated with chemotherapy response and prognosis in colorectal cancer

Cancer Cell Int. 2024 Feb 15;24(1):75. doi: 10.1186/s12935-024-03236-1.

Authors

Maria Kopsida¹, Na Liu², Angeliki Kotti¹, Jing Wang³, Lasse Jensen⁴, Ganesan Jothimani⁵, Camilla Hildesjo¹, Staffan Haapaniemi⁶, Wen Zhong³, Surajit Pathak^{7

8}, Xiao-Feng Sun⁹

Affiliations

¹ Department of Oncology and Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.
² Department of Gastroenterology, Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
³ Science for Life Laboratory, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden.
⁴ Department of Medical and Health Sciences, Linköping University, Linköping, Sweden.
⁵ Department of Medical Biotechnology, Faculty of Allied Health Sciences, Chettinad Hospital and Research Institute, Chettinad Academy of Research and Education, Kelambakkam, Tamil Nadu, India.
⁶ Department of Surgery and Department of Biomedical and Clinical Sciences, Linköping University, Norrköping, Sweden.
⁷ Department of Oncology and Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden. drsurajitpathak@care.edu.in.
⁸ Department of Medical Biotechnology, Faculty of Allied Health Sciences, Chettinad Hospital and Research Institute, Chettinad Academy of Research and Education, Kelambakkam, Tamil Nadu, India. drsurajitpathak@care.edu.in.
⁹ Department of Oncology and Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden. xiao-feng.sun@liu.se.

Abstract

Purpose: To examine the role of RhoB expression in relation to chemotherapy response, clinical outcomes and associated signaling pathways in colorectal cancer patients.

Materials and methods: The study included 5 colon cancer cell lines, zebrafish embryos and 260 colorectal cancer patients treated with 5-fluorouracil (5-FU) and oxaliplatin (OXL). The methods consisted of CRISPR/Cas9, reactive oxygen species (ROS), caspase-3 activity, autophagy flux, in-silico RNA sequencing and immunohistochemistry. Gene expression analysis and pathway analysis were conducted using RNA-seq data.

Results: All cancer lines tested, including SW480, SW480-KO13 (RhoB knockout), SW480-KO55 (RhoB knockout), HCT116 and HCT116-OE (RhoB overexpressed), exhibited cytotoxicity to 5-FU and OXL. RhoB knockout cell lines demonstrated significantly reduced migration compared to the control cell lines. Furthermore, RhoB played a role in caspase-3-dependent apoptosis, regulation of ROS production and autophagic flux. The mRNA sequencing data indicated lower expression levels of oncogenes in RhoB knockout cell lines. The zebrafish model bearing SW480-KO showed a light trend toward tumor regression. RhoB expression by immunohistochemistry in patients was increased from normal mucosa to tumor samples. In patients who received chemotherapy, high RhoB expression was related to worse survival compared to low RhoB expression. Furthermore, the molecular docking analysis revealed that OXL had a higher binding affinity for RhoB than 5-FU, with a binding affinity of -7.8 kcal/mol and HADDOCK predicted molecular interactions between RhoB and caspase 3 protein. Gene-set enrichment analysis supported these findings, showing that enrichment of DNA damage response pathway and p53 signaling in RhoB overexpression treatment group, while the RhoB knockout treatment group exhibited enrichment in the negative regulation pathway of cell migration.

Conclusion: RhoB was negatively associated with chemotherapy response and survival in colorectal cancers. Therefore, RhoB inhibition may enhance chemotherapeutic responses and patient survival.

Keywords: 5-fluorouracil; Colorectal cancer; Oxaliplatin; RhoB; Signaling pathway; Survival.