Basic Study
Copyright ©The Author(s) 2019. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastrointest Oncol. Nov 15, 2019; 11(11): 971-982
Published online Nov 15, 2019. doi: 10.4251/wjgo.v11.i11.971
Calponin 3 promotes invasion and drug resistance of colon cancer cells
Vidhya A Nair, Noura A Al-khayyal, Sivaramakrishnan Sivaperumal, Wael M Abdel-Rahman
Vidhya A Nair, Environment and Cancer Research Group, Sharjah Institute for Medical Research, University of Sharjah, Sharjah 27272, United Arab Emirates
Noura A Al-khayyal, College of Medicine and Sharjah Institute for Medical Research, University of Sharjah, Sharjah 27272, United Arab Emirates
Sivaramakrishnan Sivaperumal, Department of Biotechnology, Bharathidasan University, Tiruchirappalli 620024, India
Wael M Abdel-Rahman, Department of Medical Laboratory Sciences, College of Health Sciences and Environment and Cancer Research Group, Sharjah Institute for Medical Research, University of Sharjah, Sharjah 27272, United Arab Emirates
Author contributions: Abdel-Rahman WM supported the study, designed the project, obtained funding, contributed to the laboratory work, provided study material, analyzed and interpreted the data, and wrote the manuscript; Nair VA developed the ideas, performed the experiments, and contributed to data analysis; Al-khayyal NA performed the experiments and contributed to data analysis; Sivaperumal S provided critical insights and contributed to the data analysis and write up; All authors revised and endorsed the final draft.
Institutional review board statement: Work involving human tissue was according to ethical standards of Helsinki declaration and under the approval of the Ethics and Research Committee at the University of Sharjah.
Conflict-of-interest statement: The authors have no conflicts of interest to declare.
Data sharing statement: All relevant data were presented in the manuscript. Further information is available from the corresponding author.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Wael M Abdel-Rahman, MD, PhD, Full Professor, Department of Medical Laboratory Sciences, College of Health Sciences and Environment and Cancer Research Group, Sharjah Institute for Medical Research, University of Sharjah, Sharjah 27272, United Arab Emirates. whassan@sharjah.ac.ae
Telephone: +971-6-5057556 Fax: +971-6-5057515
Received: March 6, 2019
Peer-review started: March 8, 2019
First decision: April 15, 2019
Revised: July 27, 2019
Accepted: September 12, 2019
Article in press: September 12, 2019
Published online: November 15, 2019
Processing time: 255 Days and 15.3 Hours
Abstract
BACKGROUND

Calponin 3 (CNN3) is an actin-binding protein expressed in smooth muscle and non-smooth muscle cells. It is required for cytoskeletal rearrangement and wound healing.

AIM

To dissect the role of CNN3 in carcinogenesis with a focus on colon cancer.

METHODS

A total of 20 cancer cell lines (8 breast, 11 colon, and HeLa cervical cancer cell as a positive control for mesenchymal phenotype) and 57 formalin-fixed, paraffin-embedded sections from archived sporadic colorectal carcinomas were included in this study. CNN3 expression analysis by western blot or immunohistochemistry was followed by functional analyses. The CNN3 gene was silenced by specific small interfering RNA (commonly known as siRNA), followed by confirmation of the silencing efficiency by western blotting. Then, the silenced cells and control siRNA-transfected cells were analyzed for changes in epithelial and mesenchymal markers, invasion, and response to 5-fluoruracil treatment. We also performed proteomics analysis using a phospho-kinase array-based panel of 45 proteins.

RESULTS

CNN3 showed positive expression in 6/8 breast and 9/11 colon cancer lines and in HeLa cells. Interestingly, the colorectal adenocarcinoma line SW480 was negative, while the cell line developed from its matching lymph node metastasis (SW620) was positive for CNN3. CNN3 expression was fairly consistent with the metastatic phenotype in colon cancer because it was absent in one other colon cell line from a primary site and expressed in all others. We selected SW620 for subsequent functional analyses. CNN3-silenced SW620 cells showed a reduction in collagen invasion and loss of mesenchymal markers. CNN3 silencing caused an increase in the SW620 colon cancer cell sensitivity to 5-fluorouracil. Phospho-kinase array-based proteomics analysis showed that CNN3 silencing in SW620 reduced extracellular signal-regulated kinase, β-Catenin, mutant p53, c-Jun, and heat shock protein 60 activities but increased that of checkpoint kinase 2. CNN3 was expressed in 20/57 (35%) colon cancer cases as shown by immunohistochemistry. CNN3 was associated with a decrease in overall survival in colon cancer in silico.

CONCLUSION

These results show the involvement of CNN3 in lymph node metastasis and resistance to chemotherapy in colon cancer and suggest that significant oncogenic pathways are involved in these CNN3-related actions.

Keywords: β-Catenin; Calponin 3; Colon cancer; Epithelial-mesenchymal transition; Invasion; Metastasis

Core tip: We hypothesized that calponin 3 (CNN3) may play a role in carcinogenesis based upon its known biological functions. We showed that it is expressed in colon and breast cancer cells and is associated with the metastatic phenotype in colon cancer via upregulating mesenchymal markers. CNN3 also improves the sensitivity to chemotherapy in these tumors. We also showed that it is linked to other carcinogenic pathways such as extracellular signal-regulated kinase 1/2, β-Catenin, mutant p53, c-Jun, and heat shock protein 60 in colorectal cancer. Thus, CNN3 is a promising biomarker in colon cancer.