Differential Expression of MicroRNA-19b Promotes Proliferation of Cancer Stem Cells by Regulating the TSC1/mTOR Signaling Pathway in Multiple Myeloma.

Abstract

BACKGROUND/AIMS:

MiR-19b has been reported to be involved in several malignancies, but its role in multiple myeloma (MM) is still unknown. The objective of this study was to explore the biological mechanism of miR-19b in the progression of MM.

METHODS:

First, we performed real-time polymerase chain reaction (PCR) and Western blot to study the expression of miR-19b, tuberous sclerosis 1 (TSC1), and caspase-3 in different groups. MTT assay was performed to explore the effect of miR-19b on survival and apoptosis of cancer stem cells (CSCs). Computation analysis and luciferase assay were utilized to confirm the interaction between miR-19b and TSC1.

RESULTS:

A total of 38 participants comprising 20 subjects with MM and 18 healthy subjects as normal controls were enrolled in our study. Real-time PCR showed dramatic upregulation of miR-19b, but TSC1 was evidently suppressed in the MM group. MiR-19b overexpression substantially promoted clonogenicity and cell viability, and further inhibited apoptosis of CSCs in vitro. Furthermore, miR-19b overexpression downregulated the expression of caspase-3, which induced apoptosis. Using in silico analysis, we identified that TSC1 might be a direct downstream target of miR-19b, and this was further confirmed by luciferase assay showing that miR-19b apparently reduced the luciferase activity of wild-type TSC1 3´-UTR, but not that of mutant TSC1 3´-UTR. There was also evident decrease in TSC1 mRNA and protein in CSCs following introduction of miR-19b. Interestingly, reintroduction of TSC1 abolished the miR-19b-induced proliferation promotion and apoptosis inhibition in CSCs.

CONCLUSION:

These findings collectively suggest that miR-19b promotes cell survival and suppresses apoptosis of MM CSCs via targeting TSC1 directly, indicating that miR-19b may serve as a potential and novel therapeutic target of MM based on miRNA expression.

• Authors: Ni Wang, Xiaohua Liang, Weijian Yu, Shihang Zhou, Meiyun Fang
• Journal: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology (Cell Physiol Biochem) Vol. 50 Issue 5 Pg. 1804-1814 ( 2018) ISSN: 1421-9778 [Electronic] Germany
• PMID: 30396185 (Publication Type: Journal Article)
• Copyright: © 2018 The Author(s). Published by S. Karger AG, Basel.

Chemical References

• 3' Untranslated Regions
• Antagomirs
• MIRN19 microRNA, human
• MicroRNAs
• TSC1 protein, human
• Tuberous Sclerosis Complex 1 Protein
• Tumor Suppressor Proteins
• MTOR protein, human
• Ribosomal Protein S6 Kinases, 70-kDa
• TOR Serine-Threonine Kinases
• Caspase 3

Topics

• 3' Untranslated Regions
• Antagomirs (metabolism)
• Apoptosis
• Base Sequence
• Case-Control Studies
• Caspase 3 (metabolism)
• Cell Line, Tumor
• Cell Proliferation
• Down-Regulation
• Humans
• MicroRNAs (antagonists & inhibitors, genetics, metabolism)
• Multiple Myeloma (genetics, pathology)
• Neoplastic Stem Cells (cytology, metabolism)
• Ribosomal Protein S6 Kinases, 70-kDa (metabolism)
• Sequence Alignment
• Signal Transduction
• TOR Serine-Threonine Kinases (metabolism)
• Tuberous Sclerosis Complex 1 Protein
• Tumor Suppressor Proteins (chemistry, genetics, metabolism)