Multipronged activity of combinatorial miR-143 and miR-506 inhibits Lung Cancer cell cycle progression and angiogenesis in vitro

Sci Rep. 2018 Jul 12;8(1):10495. doi: 10.1038/s41598-018-28872-2.

Abstract

Lung cancer (LC) is the leading cause of cancer-related deaths. Downregulation of CDK1, 4 and 6, key regulators of cell cycle progression, correlates with decreased LC cell proliferation. Enforced expression of miRNAs (miRs) is a promising approach to regulate genes. Here, we study the combinatorial treatment of miR-143 and miR-506 to target the CDK1, 4/6 genes, respectively. We analyzed the differential expression of CDK genes by qPCR, and western blot, and evaluated changes in the cell cycle distribution upon combinatorial treatment. We used an antibody microarray analysis to evaluate protein expression, focusing on the cell cycle pathway, and performed RNA-sequencing for pathway analysis. The combinatorial miR treatment significantly downregulated CDK1, 4 and 6 expression, and induced a shift of the cell cycle populations, indicating a G1 and G2 cell cycle block. The two miRs induces strong cytotoxic activity, with potential synergism, and a significant Caspase 3/7 activation. We identified a strong inhibition of tube formation in the presence or absence VEGF in an in vitro angiogenesis model. Together with the pathways analysis of the RNA-sequencing data, our findings establish the combinatorial miR transfection as a viable strategy for lung cancer treatment that merits further investigation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • A549 Cells
  • Apoptosis / genetics
  • CDC2 Protein Kinase / genetics
  • Cyclin-Dependent Kinase 4 / genetics
  • Cyclin-Dependent Kinase 6 / genetics
  • Down-Regulation
  • G1 Phase Cell Cycle Checkpoints / genetics
  • G2 Phase Cell Cycle Checkpoints / genetics
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic
  • Genetic Therapy / methods*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Lung Neoplasms / blood supply
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology
  • Lung Neoplasms / therapy*
  • MicroRNAs / genetics*
  • Neovascularization, Pathologic / genetics
  • Neovascularization, Pathologic / therapy*
  • Primary Cell Culture
  • Transfection / methods
  • Tumor Microenvironment / genetics

Substances

  • MIRN143 microRNA, human
  • MIRN506 microRNA, human
  • MicroRNAs
  • CDC2 Protein Kinase
  • CDK1 protein, human
  • CDK4 protein, human
  • CDK6 protein, human
  • Cyclin-Dependent Kinase 4
  • Cyclin-Dependent Kinase 6