Huang Jie, Wu Zhou, Wang Guangxiu, Cai Yingxian, Cai Minshan, Li Yaozhang
Cancer Center, Affiliated Hospital of Guangdong Medical College, Zhanjiang, Guangdong 524001, P.R. China.
Mol Med Rep. 2015 Oct;12(4):5730-6. doi: 10.3892/mmr.2015.4169. Epub 2015 Aug 3.
Previous studies have reported the antitumor activity of N‑Myc downstream‑regulated gene 2 (NDRG2), a novel p53‑inducible gene, in several types of cancer. The present study aimed to investigate the effects of NDRG2 expression on the proliferation of a human bladder cancer cell line. NDRG2 and control green fluorescent protein (GFP) recombinant adenovirus plasmids were constructed and transfected into a bladder cancer cell line with mutant p53 (T24 cells). NDRG2 expression was analyzed using western blot analysis and immunofluorescence assay (IFA); in addition, the subcellular localization of NDRG2 was detected using a confocal microscope. The proliferation rate of cells was measured using colony formation and MTT assays. Furthermore, the cell cycle of transfected T24 cells was detected by flow cytometry. The results indicated that T24 cells expressed low levels of NDRG2 prior to infection with GFP‑NDRG2 recombinant adenovirus; by contrast, following infection, NDRG2 was primarily overexpressed in mitochondria. The proliferation rate of T24 cells was significantly reduced by NDRG2 expression (P<0.01). In addition, 82.1% of NDRG2‑expressing cells were in S‑phase, compared to 74.4% in the control virus‑infected cells (P<0.05). Furthermore, upregulation of NDRG2 induced an increase in oncosis, rather than apoptosis, in T24 cell. In conclusion, the results of the present study indicated that NDRG2 expression in mitochondria may arrest bladder cancer cells in S‑phase as well as decrease cell proliferation through inducing oncosis. It was therefore proposed that NDRG2 was not only a biomarker, but also a tumor suppressor for bladder cancer.
先前的研究报道了一种新型的p53诱导基因N- myc下游调控基因2(NDRG2)在几种癌症中的抗肿瘤活性。本研究旨在探讨NDRG2表达对人膀胱癌细胞系增殖的影响。构建了NDRG2和对照绿色荧光蛋白(GFP)重组腺病毒质粒,并将其转染到具有突变型p53的膀胱癌细胞系(T24细胞)中。使用蛋白质免疫印迹分析和免疫荧光测定(IFA)分析NDRG2表达;此外,使用共聚焦显微镜检测NDRG2的亚细胞定位。使用集落形成和MTT测定法测量细胞的增殖率。此外,通过流式细胞术检测转染的T24细胞的细胞周期。结果表明,在感染GFP-NDRG2重组腺病毒之前,T24细胞表达低水平的NDRG2;相比之下,感染后,NDRG2主要在线粒体中过表达。NDRG2表达显著降低了T24细胞的增殖率(P<0.01)。此外,表达NDRG2的细胞中有82.1%处于S期,而对照病毒感染细胞中这一比例为74.4%(P<0.05)。此外,NDRG2的上调诱导T24细胞发生胀亡而非凋亡。总之,本研究结果表明,线粒体中NDRG2的表达可能使膀胱癌细胞停滞在S期,并通过诱导胀亡来降低细胞增殖。因此推测NDRG2不仅是膀胱癌的生物标志物,也是一种肿瘤抑制因子。