Alteration in miRNA gene expression pattern in acute promyelocytic leukemia cell induced by arsenic trioxide: a possible mechanism to explain arsenic multi-target action

摘要

MicroRNAs (miRNAs) are involved in cancer pathogenesis, apoptosis, and cell growth, and these miRNAs are thought to be functional as oncogenes and/or tumor suppressors in the gene regulatory networks. We studied the potential contribution of miRNAs in acute promyelocytic leukemia (APL) cell NB4 during the apoptosis induction by arsenic trioxide (ATO). The apoptotic effects of ATO on the NB4 cell line at a pharmacological dose (2 μM) was verified using cell growth and viability assays, MTT assay, BrdU cell proliferation assay, flow cytometric analysis, and caspase-3 activity assay. miRNAs from untreated and 2 μM ATO-treated NB4 cell line were extracted, purified, and converted to complementary DNAs. Differential expressions of 88 cancer-related miRNAs were analyzed by real-time reverse transcription PCR using miRNA PCR cancer-array system. After normalizing to the average Ct value of three housekeeping genes in the array (U6, SNORD47, and SNORD48), the fold change of miRNAs was calculated in the ATO-treated cells as compared to untreated. Among the 88 cancer-focused miRNAs, 51 miRNAs were found to be differentially expressed more than 2-fold after ATO treatment. Of these, 48 miRNAs were upregulated up to 21.65-fold changes, while three miRNAs were downregulated up to 5.19-fold changes. By screening the literature, a majority of these upregulated miRNAs were found to have tumor and/or metastatic suppressors’ functions associated with cell cycle arrest and apoptosis, as well as inhibition of angiogenesis, invasion, and metastasis. Our results demonstrate that ATO, at the relevant concentration, modulate a substantial number of cancer-related miRNAs in APL cell line; most of these are known to function as a tumor and/or metastatic suppressors and have confirmed targets involved in cell cycle arrest and apoptosis. The results of this study support the hypothesis that miRNAs may play a mediatory role in eliciting the multi-target and pleiotropic action of ATO.