Wang Tengkai, Ji Rui, Liu Guanqun, Ma Beilei, Wang Zehua, Wang Qian
Department of Internal Medicine, Qilu Hospital, Shandong University, 107 West Wenhua Road, Jinan, 250012, Shandong, P.R. China.
Department of Gastroenterology, Qilu Hospital, Shandong University, Jinan, Shandong, P.R. China.
Cancer Cell Int. 2021 Nov 10;21(1):602. doi: 10.1186/s12935-021-02291-2.
Gastric cancer (GC) is one of the most common malignancies, molecular mechanism of which is still not clear. Aberrant expression of tumor-associated genes is the major cause of tumorigenesis. DBF4 is an important factor in cancers, although there is yet no report on its function and molecular mechanism in GC.
The expression of DBF4 in tumor tissues or cells of GC was detected by qRT-PCR and western blotting. Gastric cancer cell line MGC-803 and AGS were transfected with DBF4 siRNA or overexpression vector to detect the function of DBF4 in proliferation, migration and the sensitivity to 5-Fu with CCK-8 assay, colony formation assay, transwell assay, and wound healing assay. miR-30a was found to be the regulator of DBF4 by online bioinformatics software and confirmed with qRT-PCR, western blot and dual-luciferase reporter assays.
In our study, increased expression of DBF4 in GC tissues was first identified through The Cancer Genome Atlas (TCGA) and later confirmed using specimens from GC patients. Furthermore, functional experiments were applied to demonstrate that DBF4 promotes cell proliferation and migration in GC cell lines, moreover weakens the sensitivity of MGC803 and AGS cells to 5-Fu. We further demonstrated that miR-30a showed significantly lower expression in GC cells and inhibited the expression of DBF4 through 3'-UTR suppression. Furthermore, rescue experiments revealed that the miR-30a-DBF4 axis regulated the GC cell proliferation, migration and the sensitivity to 5-Fu. The important composition in tumor microenvironment, lactate, may be the primary factor that suppressed miR-30a to strengthen the expression of DBF4.
Taken together, our study was the first to identify DBF4 as a regulator of cell proliferation and migration in GC. Furthermore, our study identified the lactate-miR-30a-DBF4 axis as a crucial regulator of tumor progression and the tumor sensitivity to 5-Fu, which maybe serve useful for the development of novel therapeutic targets.
胃癌（GC）是最常见的恶性肿瘤之一，其分子机制仍不清楚。肿瘤相关基因的异常表达是肿瘤发生的主要原因。DBF4是癌症中的一个重要因子，尽管目前尚无关于其在胃癌中的功能和分子机制的报道。
采用qRT-PCR和western blotting检测DBF4在胃癌肿瘤组织或细胞中的表达。用DBF4 siRNA或过表达载体转染胃癌细胞系MGC-803和AGS，通过CCK-8法、集落形成试验、transwell试验和伤口愈合试验检测DBF4在增殖、迁移及对5-氟尿嘧啶敏感性方面的功能。通过在线生物信息学软件发现miR-30a是DBF4的调节因子，并通过qRT-PCR、western blot和双荧光素酶报告基因试验进行了验证。
在我们的研究中，首先通过癌症基因组图谱（TCGA）确定了胃癌组织中DBF4表达增加，随后使用胃癌患者的标本进行了验证。此外，功能实验表明DBF4促进胃癌细胞系的细胞增殖和迁移，并且削弱MGC803和AGS细胞对5-氟尿嘧啶的敏感性。我们进一步证明，miR-30a在胃癌细胞中表达显著降低，并通过3'-UTR抑制作用抑制DBF4的表达。此外，挽救实验表明miR-30a-DBF4轴调节胃癌细胞的增殖、迁移及对5-氟尿嘧啶的敏感性。肿瘤微环境中的重要成分乳酸可能是抑制miR-30a以增强DBF4表达的主要因素。
综上所述，我们的研究首次确定DBF4是胃癌细胞增殖和迁移的调节因子。此外，我们的研究确定了乳酸-miR-30a-DBF4轴是肿瘤进展和肿瘤对5-氟尿嘧啶敏感性的关键调节因子，这可能有助于开发新的治疗靶点。
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