miR-378a-3p 通过直接靶向 RAB31 抑制 Hedgehog 通路蛋白 GLI1/2，在胃癌中作为肿瘤抑制因子发挥作用。
MiR-378a-3p acts as a tumor suppressor in gastric cancer directly targeting RAB31 and inhibiting the Hedgehog pathway proteins GLI1/2.
机构信息
Medical School of Chinese PLA, Beijing 100853, China.
Senior Department of General Surgery, The First Medical Center, Chinese PLA General Hospital, Beijing 100853, China.
出版信息
Cancer Biol Med. 2022 Oct 18;19(12):1662-82. doi: 10.20892/j.issn.2095-3941.2022.0337.
OBJECTIVE
To improve the prognosis of patients with gastric cancer (GC), more effective therapeutic targets are urgently needed. Increasing evidence indicates that miRNAs are involved in the progression of various tumors, and RAS-associated protein in the brain 31 (RAB31) is upregulated and promotes the progression of multiple malignant tumors. Here, we focused on identifying RAB31-targeted miRNAs and elucidating their potential mechanism in the progression of GC.
METHODS
RAB31 and miR-378a-3p expression levels were detected in paired fresh GC tissues and GC cell lines. Bioinformatics analysis was used to predict the miRNAs targeting RAB31 and the relationships between RAB31 and other genes. Dual-luciferase reporter assays were applied to verify the targeted interaction relationship. CCK-8, colony formation, flow cytometry, wound healing, and Transwell assays were performed to assess the proliferation, apoptosis, migration, and invasion of GC cells. Tumorsphere formation assays were performed to assess the stemness of gastric cancer stem cells. Related proteins were detected by Western blot. Xenograft assays in nude mice were performed to explore the effect of miR-378a-3p .
RESULTS
We report the first evidence that miR-378a-3p is downregulated in GC, whereas its overexpression inhibits proliferation, invasion, and migration as well as promotes apoptosis in GC cells. Mechanistically, miR-378a-3p inhibits the progression of GC by directly targeting RAB31. Restoring RAB31 expression partially offsets the inhibitory effect of miR-378a-3p. Further research revealed that miR-378a-3p inhibits GLI1/2 in the Hedgehog signaling pathway and attenuates the stemness of gastric cancer stem cells. Finally, xenograft assays showed that miR-378a-3p inhibits GC tumorigenesis .
CONCLUSIONS
MiR-378a-3p inhibits GC progression by directly targeting RAB31 and inhibiting the Hedgehog signaling pathway proteins GLI1/2.
目的
为改善胃癌（GC）患者的预后，迫切需要更有效的治疗靶点。越来越多的证据表明，miRNAs 参与了多种肿瘤的进展，而脑蛋白 Ras 相关蛋白 31（RAB31）上调并促进了多种恶性肿瘤的进展。在这里，我们专注于鉴定 RAB31 靶向的 miRNAs，并阐明它们在 GC 进展中的潜在机制。
方法
检测配对的新鲜 GC 组织和 GC 细胞系中 RAB31 和 miR-378a-3p 的表达水平。生物信息学分析用于预测靶向 RAB31 的 miRNAs 以及 RAB31 与其他基因之间的关系。双荧光素酶报告基因实验用于验证靶向相互作用关系。CCK-8、集落形成、流式细胞术、划痕愈合和 Transwell 实验用于评估 GC 细胞的增殖、凋亡、迁移和侵袭。肿瘤球形成实验用于评估胃癌干细胞的干性。通过 Western blot 检测相关蛋白。裸鼠异种移植实验用于探索 miR-378a-3p 的作用。
结果
我们首次报道 miR-378a-3p 在 GC 中下调，而过表达 miR-378a-3p 抑制 GC 细胞的增殖、侵袭和迁移，并促进其凋亡。机制上，miR-378a-3p 通过直接靶向 RAB31 抑制 GC 的进展。恢复 RAB31 的表达部分抵消了 miR-378a-3p 的抑制作用。进一步的研究表明，miR-378a-3p 抑制 Hedgehog 信号通路中的 GLI1/2 并减弱胃癌干细胞的干性。最后，异种移植实验表明 miR-378a-3p 抑制 GC 肿瘤发生。
结论
miR-378a-3p 通过直接靶向 RAB31 抑制 GC 进展，并抑制 Hedgehog 信号通路蛋白 GLI1/2。
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