Jiang Zhou, Li Shu, Han Meng-Jiao, Hu Guo-Ming, Cheng Pu
Department of Oncology, Second Affiliated Hospital, Zhejiang University School of Medicine, Zhejiang University Hangzhou 310009, China.
Department of Hematology, Second Affiliated Hospital, Zhejiang University School of Medicine Hangzhou, China.
Am J Transl Res. 2020 Jul 15;12(7):3858-3870. eCollection 2020.
NSUN5, a gene encodes a cytosine-5 RNA methyltransferase, is rarely mentioned in cancers. Our study is the first one to evaluate the role of NSUN5 in the progression of colorectal cancer. Data from TCGA was used to show the different expression of NSUN5 between CRC tumor tissues and adjacent normal ones. The NSUN5 expression in the tissue microarray was detected by immunohistochemistry (IHC). qRT-PCR was conducted for NSUN5 expression examination in CRC cell lines. Cell proliferation was analyzed by the Celigo machine. GESA and correlation analysis were performed to reveal the possible underlying mechanism. The effects of NSUN5 expression on CRC cell behavior in vitro were analyzed by flow cytometry and β-galactosidase staining. The expression of cell-cycle related proteins were evaluated by western blot. Subcutaneously implanted tumor model was carried out for animal experiment. NSUN5 expression was up-regulated in CRC tumor tissues and cells, and associated with advanced tumor stages (III, IV). NSUN5 could promote cell proliferation, trigger cell cycle arrest in vitro and boost tumor growth in vivo. In addition, knockdown of NSUN5 could lead to a higher expression of Rb and a lower expression of CDK4, CDK6, p-Rb and CCNE1, but made no difference on P21, Bcl-2, caspase3 and C-Caspase3 of CRC cells. Taken together, we identify NSUN5 as a promoter in CRC development via cell cycle regulation.
NSUN5是一种编码胞嘧啶-5 RNA甲基转移酶的基因，在癌症中很少被提及。我们的研究是首个评估NSUN5在结直肠癌进展中作用的研究。利用来自TCGA的数据展示了结直肠癌肿瘤组织与相邻正常组织中NSUN5的表达差异。通过免疫组织化学（IHC）检测组织芯片中的NSUN5表达。对结直肠癌细胞系进行qRT-PCR检测NSUN5表达。用Celigo机器分析细胞增殖。进行基因集富集分析（GESA）和相关性分析以揭示可能的潜在机制。通过流式细胞术和β-半乳糖苷酶染色分析NSUN5表达对体外结直肠癌细胞行为的影响。通过蛋白质免疫印迹法评估细胞周期相关蛋白的表达。进行皮下植入肿瘤模型的动物实验。NSUN5在结直肠癌肿瘤组织和细胞中表达上调，并与晚期肿瘤阶段（III、IV期）相关。NSUN5可促进细胞增殖，在体外引发细胞周期阻滞并在体内促进肿瘤生长。此外，敲低NSUN5可导致结直肠癌细胞中Rb表达升高，CDK4、CDK6、p-Rb和CCNE1表达降低，但对P21、Bcl-2、caspase3和C-Caspase3无影响。综上所述，我们通过细胞周期调控确定NSUN5是结直肠癌发展的促进因子。
