Down-regulation of circ-PRKCI inhibits cell migration and proliferation in Hirschsprung disease by suppressing the expression of miR-1324 target PLCB1

Cell Cycle. 2018;17(9):1092-1101. doi: 10.1080/15384101.2018.1480210. Epub 2018 Jul 9.

Abstract

Circular RNAs (circRNAs) are a novel class of noncoding RNAs (ncRNAs), which have been shown to participate in intracellular RNA regulatory networks and play vital roles in many pathological processes. Recently, circular RNA_PRKCI (circ-PRKCI) has been reported to regulate cell proliferation, migration and invasion in several human cancers. Hirschsprung disease (HSCR) is a well-known congenital gut motility disorder which roots in the aberrance of cranial-caudal neural crest cell migration. In this study, we investigated whether circ-PRKCI may affect cell migration and proliferation in HSCR. Quantitative reverse transcription PCR (qRT-PCR) was performed to detect the expression of circ-PRKCI in 48 HSCR aganglionic tissues and 48 normal bowel tissues. Luciferase reporter assay and RNA immunoprecipitation (RIP) assay verified the direct interaction between miR-1324 and PLCB1 or circ-PRKCI. Cell counting Kit-8 (CCK-8) and Ethynyldeoxyuridine (EdU) assays were employed to appraise the effects of miR-1324 or circ-PRKCI on cell proliferative potential, while transwell was performed to detect the migration in vitro. We found that circ-PRKCI was significantly down-regulated in HSCR aganglionic tissues. Morever, knockdown of circ-PRKCI suppressed cell proliferation and migration in vitro. Mechanistically, we confirmed that circ-PRKCI functioned as a molecular sponge for miR-1324 to upregulate the expression of PLCB1. In conclusion, our present study revealed the important role of circ-PRKCI-miR-1324-PLCB1 regulatory network in HSCR, providing a novel insight for the pathogenesis of HSCR.

Keywords: Hirschsprung disease; circular RNA_PRKCI; miR-1324; migration; proliferation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cell Movement*
  • Cell Proliferation*
  • Down-Regulation
  • Female
  • Gene Silencing
  • HEK293 Cells
  • Hirschsprung Disease / pathology*
  • Humans
  • Infant
  • Isoenzymes / genetics
  • Isoenzymes / metabolism*
  • Male
  • MicroRNAs / metabolism*
  • Phospholipase C beta / metabolism*
  • Protein Kinase C / genetics
  • Protein Kinase C / metabolism*
  • RNA, Circular / metabolism*
  • ROC Curve
  • Response Elements

Substances

  • Isoenzymes
  • MIRN1324 microRNA, human
  • MicroRNAs
  • RNA, Circular
  • Protein Kinase C
  • protein kinase C lambda
  • PLCB1 protein, human
  • Phospholipase C beta

Grants and funding

This work was supported by the National Natural Science Foundation of China [81701493]; National Natural Science Foundation of China [81700449]; National Natural Science Foundation of China [81570467].