Tumour Bio：三氧化二砷诱导急性早幼粒细胞白血病细胞miRNA变化

MicroRNAs（miRNA）参与癌症的发病、细胞凋亡以及细胞生长，这些miRNAs被认为在基因调控网络发挥癌基因和/或抑癌基因的功能。

近日，杂志Tumour Biol上一则研究着重探究了在三氧化二砷（ATO）诱导急性早幼粒细胞白血病（APL）细胞NB4细胞凋亡过程中miRNA的潜在作用。研究人员用MTT法、尿嘧啶细胞增殖实验、流式细胞仪分析技术以及caspase-3活性测定实验确定ATO对NB4细胞株细胞凋亡影响的药理剂量。

提取纯化未处理的和2 muM ATO处理的NB4细胞系中miRNA，并转化为互补DNA。结果发现三氧化二砷处理细胞后，88个miRNA中的51个miRNA出现差异表达，表达量提高了2倍以上。其中，48个miRNA上调了21.65倍，而​​3个miRNAs表达下调了5.19倍。

通过文献筛选发现，大多数这些上调的miRNA与肿瘤和/或转移抑制基因、细胞周期阻滞和凋亡以及抑制血管生成、侵袭和转移相关。

研究结果表明三氧化二砷，能在一定浓度下调节APL细胞株相关miRNA，其中大部分是众所周知肿瘤和/或转移性抑制因子，并已确认参与细胞周期阻滞、细胞凋亡。这项研究的结果支持了miRNA可能在ATO治疗急性早幼粒细胞白血病中发挥关键作用这一假设。

doi:10.1007/s13277-011-0259-1
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Alteration in miRNA gene expression pattern in acute promyelocytic leukemia cell induced by arsenic trioxide: a possible mechanism to explain arsenic multi-target action.

Ghaffari SH, Bashash D, Dizaji MZ, Ghavamzadeh A, Alimoghaddam K.

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.