Berberine induced apoptosis of ht-29 cells through mediating of mir-21

Sara Tutunchi,1,* Roghayeh tofigh,2 Mina zare,3 Ghodratollah panahi,4

1. Department of Medical Genetics, Shahid Sadoughi University of Medical Sciences, Yazd, Iran
2. Department of animal biology, Tabriz University, Tabriz, Iran
3. Department of clinical biochemistry, Shiraz University of Medical Sciences, Shiraz, Iran
4. Department of clinical biochemistry, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Introduction

Berberine is an isoquinoline that can be extracted from many medicinal herbs. micrornas (mirnas) are small non-coding rnas that through post transcriptionally control gene expression. cumulating data suggest that mirnas have role in many diseases, including cancer. oncogenic function of mir-21 has a widely described in regulating cancer cell apoptosis. in this study, we investigate the mechanisms by which berberine suppressed the proliferation of ht-29 cells.

Methods

The effect of berberine (0,0.01,0.1,1,10, 50, 100, 200, and 500µm) on cell viability was assessed by mtt. mirna and total rna were extracted from ht-29 cells using the mirneasy kit (qiagen). cdna synthesis was performed using miscript ii rt kit (qiagen) and real time-pcr was performed using realq miscript sybr green pcr kit (qiagen).

Results

50 μmol/l berberine for 24 h was unaffected cell proliferation of ht-29 cells but, after treatment with a high concentration of berberine (≥50 μmol/l) the viability of ht-29 cells was significantly reduced compared to the control cells (p<0.05) (figure 1). berberine (100µm) after 24 h significantly increases caspase-8, bcl-2 and bax expression in ht-29 cells (p < 0.001) (figure 2). also, our data suggested that mir-21 was significantly downregulated (p < 0.001) after treatment with berberine (100µm) for 24 h (figure 3a).

Conclusion

The result of present study demonstrated that berberine inhibits the proliferation of ht-29 cells and increasing expression of bcl-2, bax and caspas-8. also, our study suggests that berberine could activate apoptosis in ht-29 cells. berberin-induced apoptosis in is mediated through the downregulating of mir-21.

Keywords

Colorectal cancer, mir-21, bcl-2, caspase 8, bax