MiR-145 inhibits proliferation of primary colon adenocarcinoma cells via induction of apoptosis, cell cycle arrest and inhibition of cell migration

Yong Yang,Xi Ming,Peng Li,Xiu-Tian Guo,Xu-Feng Ding

doi:10.4314/tjpr.v17i8.5

Abstract

Purpose: To investigate the role and potential of miR-145 as a therapeutic target for the treatment of primary colon adenocarcinoma in cell linesMethods: The expression of miR145 was determined by quantitative real time-polymerase chain reaction (RT-PCR), while cell viability was determined by MTT assay. Apoptosis was assessed by 4',6-diamidino-2-phenylindole (DAPI), acridine orange/ethidium bromide (AO/EB), and annexin V/PI doublestaining. Cell cycle analysis was performed by flow cytometry, while immunoblotting was used to determine protein expression.Results: The expression of miR-145 was significantly enhanced in all the colon adenocarcinoma cell lines investigated. On the other hand, suppression of miR-145 expression led to significant decrease in cell viability, activation of apoptosis, G2/M cell cycle arrest, and inhibition of migration of colon adenocarcinoma cells.Conclusion: These results indicate that miR-145 regulates the proliferation and metastasis of colon adenocarcinoma cells. Thus, it may be a prospective drug target for the treatment of this disease.Keywords: MicroRNA, Apoptosis, Cell migration, Proliferation

Highlights

Primary colon adenocarcinoma is one of the lethal cancers, and is responsible for significant level of mortality across the globe [1]
Cell proliferation is regulated by microRNAs from several types of cells [6]
The results revealed that the transcripts of miR145 were significantly upregulated in all the colon adenocarcinoma cell lines (WiDr, SW48, LS123, and LS180), when compared to the normal colon cell line (CCD-18Co) (p < 0.05, Figure 1)

Summary

INTRODUCTION

Primary colon adenocarcinoma is one of the lethal cancers, and is responsible for significant level of mortality across the globe [1]. The expressions of MiR-145 in four different colon adenocarcinoma cell lines were determined. The effect of miR145 on the proliferation, cell viability and migration of the human colon adenocarcinoma cells were investigated. Colon adenocarcinoma cells (SW48) were cultured in 6-well plates at a density of 1×106 cells per well, and incubated for 24 h. This was immediately followed by DAPI staining. Protein concentration was estimated in each cell extract, the protein samples were subjected to electrophoresis (SDS–PAGE) This was followed by transference to a nitrocellulose membrane and incubation with the primary antibody (1:1000) for 24 h at 4 ○C. Difference between the samples were considered statistically significant at p < 0.05

RESULTS

CONCLUSION

DISCUSSION