MiR-1254 inhibits proliferation, migration and invasion of human brain tumour cell lines

Gang Li,Yu-Hua Guan,Yan Wang,Hua Tang,Lai Jiang,Gang Zheng,Xin Huang,Dong-Gang Zhao

doi:10.4314/tjpr.v17i1.6

Abstract

Purpose: To investigate the expression of miR-1254 in 5 astrocytoma cell lines, and the mechanism involved.Methods: Total RNA was isolated by RNeasy RNA isolation kit while cDNA was prepared by RevertAid cDNA synthesis kit. The transcripts were analysed by real-time polymerase chain reaction (RT-PCR). Transfection of miR-1254 was carried out using FuGENE HD (Promega). Apoptosis was determined by DAPI, acridine orange (AO)/ethidium bromide (EB) and annexin V/PI double staining. Cell migration and invasion were investigated by wound healing and Martigel invasion assays, respectively. Protein expression was measured by western blotting.Results: Expression of miR-1254 was significantly down-regulated in the astrocytoma cell lines when compared to normal astrocyte cells (p < 0.05). Ectopic expression of miR-1254 in astrocytoma SW 1088 cells inhibited cell proliferation via initiation of apoptosis and cell cycle arrest. Over-expression of miR- 1254 also led to significant decrease in cell migration and invasion of SW 1088 astrocytoma cells (p < 0.05).Conclusion: The results show that the expression of miR-1254 is down-regulated in astrocytoma cell lines, but over-expression of miR-1254 inhibits proliferation of the cell lines via cell cycle arrest and apoptosis. Thus, miR-1254 has promising potential for use in the treatment of brain tumour.Keywords: Brain tumour, Astrocytoma, miR-1254, Apoptosis, Cell migration

Highlights

Micro RNAs are small, non-coding RNA molecules that play vital roles in gene silencing
The expression of Micro RNAs (miRNAs) may be up-regulated in one cancer type, but it may be down-regulated in other type [4]
Cultured SW 1088 cells transfected with pMSCv-pur vector containing miR-1254 or empty vector were seeded at a density of 1.2 × 104 to 1.5 × 104 in 96-well microtitre plates

Summary

INTRODUCTION

Micro RNAs (miRNAs) are small, non-coding RNA molecules that play vital roles in gene silencing. The present study was carried out to determine the expression of miR-1254 in five different astrocytoma cell lines and in normal human astrocytes. The expression profile of miR-1254 was studied using quantitative RT-PCR in the five astrocytoma cell lines (CCF-STTG1, SW 1088, SW 178, U-87 M and U-118 M) and normal human astrocytes. Cultured SW 1088 cells transfected with pMSCv-pur vector containing miR-1254 or empty vector were seeded at a density of 1.2 × 104 to 1.5 × 104 in 96-well microtitre plates. This was followed by the addition of MTT solution to the wells. The SW1088 brain tumour astrocytoma cells were subjected to annexin V/PI and examined by flow cytometry

RESULTS

CONCLUSION

Conflict of interest