Department of Dermatology, Yantai Yuhuangding Hospital, Yantai, China.
Department of Dermatology, Yantai Yeda Hospital, Yantai, China.
Neoplasma. 2019 Nov;66(6):908-917. doi: 10.4149/neo_2018_181205N923. Epub 2019 Jun 28.
miRNA has shown its potential in the regulation of squamous cell carcinoma (SCC). However, the mechanism of such an effect was not quite clear. Therefore, we aimed to investigate whether miR-30a-5p participated in the regulation of cutaneous SCC (cSCC) and the possible mechanism involved. 5-Ethynyl-2'-deoxyuridine (EdU) and cell cycle were measured using flow cytometry. The formation of cell colony was tested by colony formation assay. The capacities of migration and invasion were tested by wound healing assay and Transwell invasion assay, respectively. The target of miR-30a-5p was predicted by bioinformatics and identified by luciferase assay. Western blot was used for the determination of proteins and qPCR was for mRNA levels. miR-30a-5p expression was lowered in SCL-1 and A431 cells, and its upregulation suppressed EdU positive cells, colony numbers, migration, invasion and Bcl-2 expression, and elevated Bcl-2-associated X protein (Bax) and cleaved Caspase-3 expressions, arresting cell cycle in G1 phase. Moreover, forkhead box protein G1 (FOXG1) was proved to be the target of miR-30a-5p, and FOXG1 overexpression partially offsets the decreased colony numbers, migration and invasion rates due to miR-30a-5p overexpression in SCL-1 and A431 cells. miR-30a-5p showed a regulatory role on the expression of FOXG1 and further modulated the progressing of cSCC cells, which could be a novel pathway intervening the development of cSCC.
miRNA 在调节鳞状细胞癌（SCC）方面显示出其潜力。然而，这种作用的机制尚不清楚。因此，我们旨在研究 miR-30a-5p 是否参与调节皮肤鳞状细胞癌（cSCC）及其可能涉及的机制。通过流式细胞术测量 5-乙炔基-2'-脱氧尿苷（EdU）和细胞周期。通过集落形成测定法测试细胞集落的形成。通过划痕愈合测定法和 Transwell 侵袭测定法分别测试迁移和侵袭能力。通过生物信息学预测 miR-30a-5p 的靶标，并通过荧光素酶测定法进行鉴定。Western blot 用于测定蛋白质，qPCR 用于测定 mRNA 水平。miR-30a-5p 在 SCL-1 和 A431 细胞中的表达降低，其上调抑制 EdU 阳性细胞、集落数量、迁移、侵袭和 Bcl-2 表达，并提高 Bcl-2 相关 X 蛋白（Bax）和裂解 Caspase-3 的表达，将细胞周期阻滞在 G1 期。此外，叉头框蛋白 G1（FOXG1）被证明是 miR-30a-5p 的靶标，FOXG1 过表达部分抵消了由于 miR-30a-5p 过表达导致 SCL-1 和 A431 细胞中集落数量、迁移和侵袭率降低的情况。miR-30a-5p 对 FOXG1 的表达具有调节作用，并进一步调节 cSCC 细胞的进展，这可能是干预 cSCC 发展的新途径。
