Najafi Zoya, Sharifi Mohammadreza, Javadi Gholamreza
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran.
Department of Genetics and Molecular Biology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
J Gastrointest Cancer. 2020 Mar;51(1):109-115. doi: 10.1007/s12029-019-00215-y.
Dysregulation of microRNAs (miRNAs) has been shown to be involved in the pathogenesis and progression of many malignancies. Human hepatocellular carcinoma (HCC) is one of the most common cancers worldwide and the third cause of cancer-related deaths. Recent data suggest that microRNA-23b (miR-23b) is significantly high in different types of cancer, specifically human hepatocellular carcinoma. Locked nucleic acid (LNA)-modified oligonucleotides have recently been suggested as a novel approach for targeting miRNAs as antisense-based gene silencing. The aim of this study was to explore the functional role of LNA-anti-miR-23b in a HepG2 (hepatocarcinoma) cell line.
HepG2 cells were transfected with LNA-anti-miR-23b for 24, 48, and 72 h. Quantitative real-time reverse transcriptase-PCR (qRT-PCR) was performed to assess miR-23b expression by LNA-anti-miR-23b. The viability of the cells was evaluated by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay.
LNA-anti-miR-23b was successfully transfected into human HepG2 cells and suppressed the miR-23b. LNA-anti-miR-23b reduced the invasive behaviors of HepG2 cells after 24 h, compared to untreated cells and scrambled LNA-transfected cells, and this effect was more pronounced after 72 h.
Our findings suggest that inhibition of miR-23b could be used as a novel approach in inhibition of HCC proliferation.
微小RNA（miRNA）失调已被证明与许多恶性肿瘤的发病机制和进展有关。人类肝细胞癌（HCC）是全球最常见的癌症之一，也是癌症相关死亡的第三大原因。最近的数据表明，微小RNA-23b（miR-23b）在不同类型的癌症中显著升高，特别是在人类肝细胞癌中。锁核酸（LNA）修饰的寡核苷酸最近被认为是一种靶向miRNA的新型方法，可作为基于反义的基因沉默。本研究的目的是探讨LNA-抗miR-23b在HepG2（肝癌）细胞系中的功能作用。
将LNA-抗miR-23b转染HepG2细胞24、48和72小时。进行定量实时逆转录PCR（qRT-PCR）以评估LNA-抗miR-23b对miR-23b表达的影响。通过MTT（3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑）试验评估细胞活力。
LNA-抗miR-23b成功转染到人HepG2细胞中并抑制了miR-23b。与未处理的细胞和乱序LNA转染的细胞相比，LNA-抗miR-23b在24小时后降低了HepG2细胞的侵袭行为，并且在72小时后这种作用更加明显。
我们的研究结果表明，抑制miR-23b可作为抑制HCC增殖的一种新方法。
