Wang Zuopeng, Yao Wei, Li Kai, Zheng Na, Zheng Chao, Zhao Xiaolong, Zheng Shan
Department of Pediatric Surgery, Children's Hospital of Fudan University, Shanghai 201102, P.R. China.
Department of Endocrinology, Huashan Hospital of Fudan University, Shanghai 200040, P.R. China.
Oncol Lett. 2017 Jun;13(6):4727-4733. doi: 10.3892/ol.2017.6052. Epub 2017 Apr 20.
MicroRNA (miR/miRNA)-21 is a well-known oncogenic miRNA that is overexpressed in various types of tumors. The tumor-suppressor genes programmed cell death 4 (PDCD4) and phosphatase tensin homologue (PTEN), are targets of miR-21, and are underexpressed in several types of cancer. However, the expression of miR-21 and its target genes in neuroblastoma (NB) remains unclear. In the present study, a miR-21 inhibitor oligonucleotide was transfected into the SK-N-SH cell line, and the expression of miR-21, PTEN and PDCD4 was detected through quantitative polymerase chain reaction analysis. Western blotting was used to examine levels of PTEN, PDCD4 and caspase-3 proteins. The expression of PTEN and PDCD4 in the SK-N-SH cell line transfected with the miR-21 inhibitor was significantly increased compared with untransfected SK-N-SH and negative control-transfected cells. Cell proliferation was inhibited and the apoptotic ratio was significantly increased in miR-21 inhibitor-transfected cells compared with untransfected SK-N-SH and negative control-transfected cells. Western blot analysis revealed a significant increase in caspase-3 expression compared with untransfected SK-N-SH and negative control-transfected cells. The results of the present study indicate that miR-21 may serve an oncogenic role in the cellular processes underlying NB development and thus may be a novel therapeutic target for the treatment of patients with NB.
微小RNA(miR/miRNA)-21是一种著名的致癌性微小RNA,在多种类型的肿瘤中均有过表达。肿瘤抑制基因程序性细胞死亡4(PDCD4)和张力蛋白磷酸酶同源物(PTEN)是miR-21的靶标,在几种类型的癌症中表达不足。然而,miR-21及其靶标基因在神经母细胞瘤(NB)中的表达仍不清楚。在本研究中,将miR-21抑制剂寡核苷酸转染到SK-N-SH细胞系中,并通过定量聚合酶链反应分析检测miR-21、PTEN和PDCD4的表达。采用蛋白质免疫印迹法检测PTEN、PDCD4和半胱天冬酶-3蛋白的水平。与未转染的SK-N-SH细胞和阴性对照转染细胞相比,转染miR-21抑制剂的SK-N-SH细胞系中PTEN和PDCD4的表达显著增加。与未转染的SK-N-SH细胞和阴性对照转染细胞相比,转染miR-21抑制剂的细胞中细胞增殖受到抑制,凋亡率显著增加。蛋白质免疫印迹分析显示,与未转染的SK-N-SH细胞和阴性对照转染细胞相比,半胱天冬酶-3的表达显著增加。本研究结果表明,miR-21可能在NB发生的细胞过程中发挥致癌作用,因此可能是治疗NB患者的新治疗靶点。