PIK3C2A is a gene-specific target of microRNA-518a-5p in imatinib mesylate-resistant gastrointestinal stromal tumor

Autor: Jiaqian Lin, Xiaodong Gao, Chen Xu, Kuntang Shen, Qin Hu, Yalan Liu, Yuan Shi, Anwei Xue, Shaohua Lu, Jian-Fang Xu, Yingyong Hou, Yunshan Tan, Xiaojing Li, Dongxian Jiang
Rok vydání: 2016
Předmět:
Adult
Male
0301 basic medicine
Pathology
medicine.medical_specialty
Stromal cell
Gastrointestinal Stromal Tumors
Down-Regulation
Antineoplastic Agents
Apoptosis
Pathology and Forensic Medicine
Phosphatidylinositol 3-Kinases
03 medical and health sciences
0302 clinical medicine
Cell Line
Tumor
microRNA
Humans
Medicine
RNA
Neoplasm
Stromal tumor
3' Untranslated Regions
Protein Kinase Inhibitors
neoplasms
Molecular Biology
Aged
Cell Proliferation
Gastrointestinal Neoplasms
Regulation of gene expression
business.industry
Microarray analysis techniques
Cell growth
Imatinib
Cell Biology
Middle Aged
Gene Expression Regulation
Neoplastic
MicroRNAs
HEK293 Cells
030104 developmental biology
Imatinib mesylate
Drug Resistance
Neoplasm
030220 oncology & carcinogenesis
Imatinib Mesylate
Cancer research
Female
business
medicine.drug
Zdroj: Laboratory Investigation. 96:652-660
ISSN: 0023-6837
DOI: 10.1038/labinvest.2015.157
Popis: Imatinib mesylate resistance occurs in some patients with gastrointestinal stromal tumors (GISTs) during the course of treatment. In this study, we investigated the relationship between microRNAs (miRNAs) and imatinib-resistant GISTs, and the effect of miR-518a-5p on PIK3C2A in imatinib-resistant GISTs. A total of 20 matched-pair GIST samples from imatinib-resistant patients were included in the study. Each of the paired tumor specimens were from the same patient who had surgical removal of GISTs preimatinib and postimatinib treatment. Seven pairs of tissues were resected for microarray analysis, and the remaining 13 pairs were utilized for miRNAs analysis. Target genes were selected based on bioinformatics from multiple biological databases. Luciferase reporter assays were used to confirm the binding of miR-518a-5p to PIK3C2A 3'UTR. GIST882R-NC, 882R-miR-518a-5p-OE, and 882R-miR-518a-5p-KD cell lines were constructed using lentiviral vectors. miR-518a-5p and PIK3C2A expression in 882R-NC, 882R-OE, and 882R-KD cells was assessed by real-time PCR and western blotting. A cell counting kit was used to detect the influence of miR-518a-5p to cell proliferation. TUNEL staining was applied to detect the influence of miR-518a-5p to cell apoptosis. Microarray analysis showed that miR-518a-5p was downregulated in imatinib-resistant GISTs, and the expression of miR-518a-5p was confirmed with good concordance between real-time PCR and miRNA microarray results. Luciferase reporter assays indicated that miR-518a-5p bound to the PIK3C2A 3'UTR. Compared with 882R-OE, PIK3C2A expression was significantly increased in 882R-KD cells. MiR-518a-5p reduced 882R proliferation and promoted 882R apoptosis. In conclusion, PIK3C2A is a gene-specific target of miR-518a-5p in imatinib mesylate-resistant GISTs. Low expression of miR-518a-5p is likely to upregulate PIK3C2A and affect the cellular response to the drug, causing resistance to imatinib in GISTs.
Databáze: OpenAIRE
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