Abstract
miRNAs play an important regulatory role in variety of cellular functions and several diseases, including cancer. MicroRNA-21 (miR-21) is overexpressed in almost all types of human cancers. Studies revealed that the knockdown of miR-21 results in reduced tumor cell growth, cell cycle arrest and cell apoptosis. In this study, we evaluated the effect of doxorubicin on miR-21 expression in mcf-7 breast cancer cells. miRNA was extracted from mcf-7 cells treated with doxorubicin and untreated cells using miRNeasy Kit (Qiagen) according to the manufacturer’s instructions. cDNA synthesis was performed using miScript II RT Kit (Qiagen) and Real Time-PCR was performed using Real Q Plus 2x Master Mix Green-(Ampliqon, Denmark). The relative expression of miR-16 and miR-21 was calculated using comparative Ct method. All tests were run in triplicate to minimize the experimental errors. Samples with a Ct > 37 were excluded from the analysis. Statistically, a significant decrease in cell proliferation of mcf-7 cells was found in doxorubicin group compared with control groups 24 hours after transfection, dose dependently (p value< 0.001). After 24 hours, Doxorubicin (100 μm) significantly decreased miR-21 expression in mcf-7 cells (p = 0.0001). Also, the expression of caspase 9 significantly increased after Doxorubicin (100 μm) treatment (p = 0.0003). Together, these findings indicate that miR-21 plays a key role in regulating cell apoptosis in mcf-7 cells and may serve as a target for effective therapies.
Highlights
Cancers are a group of disease with abnormal cell growth and potential to spread to different tissues of body [1]-[6] Despite much progress in the management of cancer, cancer is still a major public health problem and one of the deadliest diseases worldwide, with approximately 14 million new cases and 8.2 million death each year [7] [8]
We evaluated the effect of doxorubicin on miR-21 expression in mcf-7 breast cancer cells. miRNA was extracted from mcf-7 cells treated with doxorubicin and untreated cells using miRNeasy Kit (Qiagen) according to the manufacturer’s instructions. cDNA synthesis was performed using miScript II RT Kit (Qiagen) and Real Time-PCR was performed using Real Q Plus 2x Master Mix Green-(Ampliqon, Denmark)
We evaluated the effect of doxorubicin on miR-21expression in mcf-7 breast cancer cells
Summary
Cancers are a group of disease with abnormal cell growth and potential to spread to different tissues of body [1]-[6] Despite much progress in the management of cancer, cancer is still a major public health problem and one of the deadliest diseases worldwide, with approximately 14 million new cases and 8.2 million death each year [7] [8]. Many studies were interested to identify specific molecules involved in breast cancer and understand their characteristics [10]. MiRNAs are negative regulators of gene expression which induce mRNA degradation or translational repression [16]. It was shown that many miRNAs are influential in the development of many human cancers [17]. Identified in many types of tumors, miRNAs can act as oncogenic or tumor suppressors [19] Dysregulation of miRNA expression has been implicated in estrogen-related diseases including breast cancer and endometrial cancer [20]. The important target genes of miR-21 are involved in cell proliferation, activation, and apoptosis [21]. We evaluated the effect of doxorubicin on miR-21expression in mcf-7 breast cancer cells
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