Department of Gastroenterology, Division of Southern Building, Chinese PLA General Hospital, Beijing 100853, P.R. China.
Oncol Rep. 2017 Aug;38(2):1133-1139. doi: 10.3892/or.2017.5758. Epub 2017 Jun 27.
Previous studies have shown that microRNA-31 (miR-31) functions as a tumor-suppressor in various types of cancer. In the present study we found that miR-31 was significantly downregulated in gastric cancer (GC) as determined by microRNA (miRNA) array screening analysis. Although aberrant expression of miR-31 has been found in different types of cancer, its pathophysiological effect and role in tumorigenesis still remain to be elucidated. In the present study, we detected miR-31 expression in both metastatic GC cell lines and tissues that are potentially highly metastatic by real-time polymerase chain reaction (PCR). Transwell and scratch healing assays were conducted to examine whether the ectopic expression of miR-31 could modify the invasion and migration abilities of GC cells in vitro. We found that miR-31 inhibited GC metastasis in a nude mouse xenograft model of GC. Luciferase reporter assays demonstrated that miR-31 could directly bind to the 3' untranslated region of RhoA and downregulate the expression of RhoA. Significant downregulation of miR-31 in 78 GC tissues and four GC cell lines was examined by real-time reverse transcription-PCR. Moreover, the decreased expression of miR-31 was demonstrated to be associated with lymph node metastasis, poor pT and pN stage, and invasion ability into lymphatic vessels as determined by the Mann-Whitney U test. We also found that miR-31 could inhibit cell invasion and migration abilities in vitro using the Transwell and scratch healing assays in BGC-823, SGC-7901, MGC-803 as well as AGS cells. Experiments in a nude mouse model revealed that miR-31 suppressed tumorigenicity in vivo. The luciferase activity assay and western blotting indicated that RhoA was the potential target of miR-31 in GC cells. Collectively, our results provide important evidence that the downregulation of miR-31 inhibited the invasion and migration abilities of GC cells through direct targeting of the tumor metastasis‑associated gene, RhoA. These findings suggest that miR-31 may be a promising therapeutic candidate in the treatment of GC metastasis.
先前的研究表明,microRNA-31 (miR-31) 在多种类型的癌症中作为肿瘤抑制因子发挥作用。在本研究中,我们通过 microRNA (miRNA) 阵列筛选分析发现 miR-31 在胃癌 (GC) 中显著下调。尽管 miR-31 的异常表达已在不同类型的癌症中被发现,但它在肿瘤发生中的病理生理作用和作用仍有待阐明。在本研究中,我们通过实时聚合酶链反应 (PCR) 检测了具有潜在高转移性的转移性 GC 细胞系和组织中的 miR-31 表达。Transwell 和划痕愈合试验用于检测 miR-31 的异位表达是否可以改变 GC 细胞在体外的侵袭和迁移能力。我们发现 miR-31 抑制了 GC 在 GC 裸鼠异种移植模型中的转移。荧光素酶报告基因试验表明,miR-31 可以直接结合 RhoA 的 3'非翻译区并下调 RhoA 的表达。通过实时逆转录-PCR 检测到 78 个 GC 组织和四个 GC 细胞系中 miR-31 的显著下调。此外,通过 Mann-Whitney U 检验,发现 miR-31 的表达降低与淋巴结转移、较差的 pT 和 pN 分期以及淋巴管侵袭能力有关。我们还发现,通过 Transwell 和划痕愈合试验,miR-31 可以抑制 BGC-823、SGC-7901、MGC-803 和 AGS 细胞中的体外细胞侵袭和迁移能力。裸鼠模型实验表明,miR-31 抑制了体内肿瘤的发生。荧光素酶活性测定和 Western blot 分析表明,RhoA 是 GC 细胞中 miR-31 的潜在靶标。总之,我们的研究结果提供了重要证据,表明 miR-31 通过直接靶向肿瘤转移相关基因 RhoA,抑制了 GC 细胞的侵袭和迁移能力。这些发现表明,miR-31 可能是治疗 GC 转移的有前途的治疗候选物。