Wang Danping, Luo Lin, Guo Junming
Zhejiang Provincial Key Laboratory of Pathophysiology, Department of Biochemistry and Molecular Biology, Ningbo University School of Medicine, 818 Fenghua Road, Ningbo, 315211, China.
Med Oncol. 2014 Aug;31(8):98. doi: 10.1007/s12032-014-0098-1. Epub 2014 Jul 10.
MicroRNAs (miRNAs) are a class of small noncoding RNAs, which regulate gene expression in the posttranscriptional level. They are involved in cancer occurrence and development. Different members of the same miRNA family may have different roles. Since the fact that metastasis is the main cause of cancer-related death and miR-129 has three members, in this study, we focused on the migration inhibitory role of miR-129-1-3p in gastric cancer and explored the possible mechanisms. We first compared the expression of three miR-129 family members, miR-129-5p, miR-129-1-3p, and miR-129-2-3p, between gastric carcinoma tissues and surgical margin non-cancer samples by quantitative real-time reverse transcription-polymerase chain reaction (QRT-PCR). Then, we selected miR-129-1-3p for further analysis and transfected its mimic and inhibitor into gastric cancer BGC-823 cells, respectively. Then, we compared the BGC-823 cells' migration capacity by transwell assay. Finally, we predicted the possible targets of miR-129-1-3p and selected one target for further analysis by QRT-PCR and luciferase reporter assay. The results showed that the expression of miR-129-1-3p in gastric carcinoma was significantly lower than that in surgical margin samples. And miR-129-1-3p could inhibit the migration of BGC-823 cells. From the candidates, we selected bradykinin receptor B2 (BDKRB2), which was reported relating to metastasis, as a target for further analysis. QRT-PCR showed that the expression of BDKRB2 was negatively related to miR-129-1-3p. Luciferase reporter assay showed that BDKRB2 was the target of the miR-129-1-3p. In summary, miR-129-1-3p inhibits the BGC-823 cell migration by targeting BDKRB2.
微小RNA（miRNA）是一类小的非编码RNA，其在转录后水平调节基因表达。它们参与癌症的发生和发展。同一miRNA家族的不同成员可能具有不同的作用。由于转移是癌症相关死亡的主要原因，且miR-129有三个成员，因此在本研究中，我们聚焦于miR-129-1-3p在胃癌中的迁移抑制作用，并探索其可能的机制。我们首先通过定量实时逆转录-聚合酶链反应（QRT-PCR）比较了胃癌组织与手术切缘非癌样本中三个miR-129家族成员miR-129-5p、miR-129-1-3p和miR-129-2-3p的表达。然后，我们选择miR-129-1-3p进行进一步分析，并分别将其模拟物和抑制剂转染到胃癌BGC-823细胞中。接着，我们通过Transwell实验比较BGC-823细胞的迁移能力。最后，我们预测了miR-129-1-3p的可能靶标，并通过QRT-PCR和荧光素酶报告基因实验选择了一个靶标进行进一步分析。结果显示，miR-129-1-3p在胃癌中的表达明显低于手术切缘样本中的表达。并且miR-129-1-3p可以抑制BGC-823细胞的迁移。从候选物中，我们选择了与转移相关的缓激肽受体B2（BDKRB2）作为进一步分析的靶标。QRT-PCR显示BDKRB2的表达与miR-129-1-3p呈负相关。荧光素酶报告基因实验表明BDKRB2是miR-129-1-3p的靶标。综上所述，miR-129-1-3p通过靶向BDKRB2抑制BGC-823细胞迁移。
