Soltani-Sedeh Hadis, Irani Shiva, Mirfakhraie Reza, Soleimani Masoud
Department of Biology, Science and Research Branch, Islamic Azad University; Stem Cell Technology Research Center, Tehran, Iran.
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
J Cancer Res Ther. 2019 Jan-Mar;15(1):32-37. doi: 10.4103/jcrt.JCRT_267_17.
MicroRNAs are small noncoding RNAs which modulate gene expression at different levels. It has been shown that downregulation of miR-34a occurs in varieties of cancers including colorectal cancer (CRC). In this study, we investigated the potential tumor inhibitory effects of miR-34a alone or in combination with paclitaxel in CRC cells.
SW480 cells were transduced with lentiviral overexpressed miR-34a. First, using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay, the effect of miR-34a induction alone or in combination with paclitaxel on the cell viability and cell proliferation were estimated. Then, the expression level of target genes was measured using quantitative reverse transcription-polymerase chain reaction analysis. Eventually, the role of miR-34a and paclitaxel on cell cycle were determined with flow cytometry.
Gene expression analysis showed that miR-34a downregulates the expression of BCL2 and SIRT1 genes at mRNA level. Furthermore, miR-34a has a potential to reduce cell viability and cell cycle arrest at G1 phase. Combination of paclitaxel with overexpression of miR-34a significantly decreased cell viability compared to cell treated with miR-34a or paclitaxel alone. Interestingly, a combination of miR-34a and paclitaxel arrested cell cycle at two phases.
Our results suggested that combination therapy of miR-34a and paclitaxel could be considered as the potential treatment of CRC.
微小RNA是一类小的非编码RNA，可在不同水平调节基因表达。研究表明，包括结直肠癌（CRC）在内的多种癌症中均存在miR-34a的下调。在本研究中，我们调查了单独使用miR-34a或与紫杉醇联合使用对CRC细胞的潜在肿瘤抑制作用。
用慢病毒过表达miR-34a转导SW480细胞。首先，使用3-（4,5-二甲基-2-噻唑基）-2,5-二苯基-2H-四氮唑溴盐法，评估单独诱导miR-34a或与紫杉醇联合使用对细胞活力和细胞增殖的影响。然后，使用定量逆转录-聚合酶链反应分析测量靶基因的表达水平。最终，通过流式细胞术确定miR-34a和紫杉醇对细胞周期的作用。
基因表达分析表明，miR-34a在mRNA水平下调BCL2和SIRT1基因的表达。此外，miR-34a具有降低细胞活力并使细胞周期停滞在G1期的潜力。与单独用miR-34a或紫杉醇处理的细胞相比，紫杉醇与miR-34a过表达联合使用可显著降低细胞活力。有趣的是，miR-34a和紫杉醇的组合使细胞周期停滞在两个阶段。
我们的结果表明，miR-34a和紫杉醇联合治疗可被视为CRC的潜在治疗方法。
