Induction, modulation and potential targets of miR-210 in pancreatic cancer cells.
BACKGROUND
MiR-210 is induced by hypoxia and plays different roles in the development of certain cancers. However, little is known about its role in pancreatic cancer (PC). This study aimed to explore the induction and modulation of PC by miR-210 and its potential molecular targets.


METHODS
PC cells were cultured under normoxic and hypoxic conditions. Expression of miR-210 and hypoxia-inducible factor (HIF)-1alpha was detected using quantitative reverse-transcription polymerase chain reaction. Cancer cells were transiently transfected with HIF-1alpha small interfering RNA (siRNA) and miR-210 mimics, and cell proliferation was measured using the CCK-8 assay. Potential targets for miR-210 were then identified using a dual luciferase reporter assay.


RESULTS
Hypoxic conditions induced miR-210 expression in six PC cell lines (AsPC-1, BxPC-3, MIAPaCa-2, PANC-1, Su86.86 and SW1990), but not in Capan-1 or T3M4 cells. Transfection of HIF-1alpha siRNA into PANC-1 cells markedly inhibited HIF-1alpha expression, and subsequently down-regulated miR-210 expression under hypoxic conditions. MiR-210 had no observable impact on the proliferation of PANC-1 or Su86.86 cells and dual luciferase reporter assays showed significantly reduced luciferase activity in the wild-type E2F3, EFNA3, GIT2, MNT, ZNF462 and EGR3 constructs, compared to the corresponding mutants, but not in HOXA3.


CONCLUSIONS
These results suggest that miR-210 expression in PC cells is induced by hypoxia through a HIF-1alpha-dependent pathway, but does not influence PC cell proliferation. Also, E2F3, EFNA3, GIT2, MNT, ZNF462 and EGR3 may be potential miR-210 targets in PC.
Department of General Surgery, Chinese Academy of Medical Sciences, Beijing, China.
Wei-Yun
WY
Chen
Wen-Jing
WJ
Liu
Yu-Pei
YP
Zhao
Li
L
Zhou
Tai-Ping
TP
Zhang
Ge
G
Chen
Hong
H
Shu
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors / Q000235:genetics / Q000378:metabolism; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D050699:E2F3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D051776:Early Growth Response Protein 3 / Q000235:genetics / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000235:genetics / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D017930:Genes, Reporter; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000235:genetics / Q000378:metabolism; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D035683:MicroRNAs / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D051778:Basic Helix-Loop-Helix Leucine Zipper Transcription Factors; D050699:E2F3 Transcription Factor; C098319:E2F3 protein, human; C079945:EGR3 protein, human; C430213:GIT2 protein, human; D020690:GTPase-Activating Proteins; C497442:HIF1A protein, human; C489832:HOXA3 protein, human; D018398:Homeodomain Proteins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; C543800:MIRN210 microRNA, human; C106465:MNT protein, human; D035683:MicroRNAs; D012097:Repressor Proteins; D051776:Early Growth Response Protein 3; D008156:Luciferases