In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells

Jung-Chien Chen; Yao-Yu Hsieh; Hsiang-Ling Lo; Albert Li; Chia-Jung Chou; Pei-Ming Yang

doi:10.3390/biom9090467

In Vitro and In Silico Mechanistic Insights into miR-21-5p-Mediated Topoisomerase Drug Resistance in Human Colorectal Cancer Cells

Biomolecules. 2019 Sep 9;9(9):467. doi: 10.3390/biom9090467.

Authors

Jung-Chien Chen^{1

2

3}, Yao-Yu Hsieh^{1

4

5}, Hsiang-Ling Lo^{1

6}, Albert Li^{1

6

7}, Chia-Jung Chou^{1

6

8}, Pei-Ming Yang^{9

10

11

12}

Affiliations

¹ PhD Program for Cancer Molecular Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University and Academia Sinica, Taipei 111, Taiwan.
² Department of Surgery, Min-Sheng General Hospital, Taoyuan 168, Taiwan.
³ Central Clinic and Hospital, Taipei 106, Taiwan.
⁴ Division of Hematology and Oncology, Shuang Ho Hospital, Taipei Medical University, New Taipei City 235, Taiwan.
⁵ Division of Hematology and Oncology, Department of Internal Medicine, School of Medicine, College of Medicine, Taipei Medical University, Taipei 111, Taiwan.
⁶ Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei 111, Taiwan.
⁷ School of Medicine, Taipei Medical University, Taipei 111, Taiwan.
⁸ TMU Research Center of Cancer Translational Medicine, Taipei 111, Taiwan.
⁹ PhD Program for Cancer Molecular Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University and Academia Sinica, Taipei 111, Taiwan. [email protected].
¹⁰ Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei 111, Taiwan. [email protected].
¹¹ TMU Research Center of Cancer Translational Medicine, Taipei 111, Taiwan. [email protected].
¹² Cancer Center, Wan Fang Hospital, Taipei Medical University, Taipei 111, Taiwan. [email protected].

Abstract

Although chemotherapy for treating colorectal cancer has had some success, drug resistance and metastasis remain the major causes of death for colorectal cancer patients. MicroRNA-21-5p (hereafter denoted as miR-21) is one of the most abundant miRNAs in human colorectal cancer. A Kaplan-Meier survival analysis found a negative prognostic correlation of miR-21 and metastasis-free survival in colorectal cancer patients (The Cancer Genome Atlas Colon Adenocarcinoma/TCGA-COAD cohort). To explore the role of miR-21 overexpression in drug resistance, a stable miR-21-overexpressing clone in a human DLD-1 colorectal cancer cell line was established. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) cell viability assay found that miR-21 overexpression induced drug resistance to topoisomerase inhibitors (SN-38, doxorubicin, and etoposide/VP-16). Mechanistically, we showed that miR-21 overexpression reduced VP-16-induced apoptosis and concomitantly enhanced pro-survival autophagic flux without the alteration of topoisomerase expression and activity. Bioinformatics analyses suggested that miR-21 overexpression induced genetic reprogramming that mimicked the gene signature of topoisomerase inhibitors and downregulated genes related to the proteasome pathway. Taken together, our results provide a novel insight into the role of miR-21 in the development of drug resistance in colorectal cancer.

Keywords: Connectivity Map; autophagy; colorectal cancer; drug resistance; microRNA.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Autophagy / drug effects
Cell Line, Tumor
Colorectal Neoplasms / pathology*
Computer Simulation*
DNA Topoisomerases / metabolism
Drug Resistance, Neoplasm / genetics*
Gene Expression Regulation, Neoplastic / drug effects
Humans
MicroRNAs / genetics*
Topoisomerase Inhibitors / pharmacology*

Substances

MIRN21 microRNA, human
MicroRNAs
Topoisomerase Inhibitors
DNA Topoisomerases