microRNA-374 通过靶向 P53 信号通路调控 Gadd45a 对人 SCC 细胞增殖、迁移、侵袭和凋亡的影响。
Effects of microRNA-374 on proliferation, migration, invasion, and apoptosis of human SCC cells by targeting Gadd45a through P53 signaling pathway.
机构信息
Department of Dermatology, Affiliated Hospital of Hebei Engineering University, Handan 056002, P.R. China.
Department of Dermatology, Affiliated Hospital of Hebei Engineering University, Handan 056002, P.R. China
出版信息
Biosci Rep. 2017 Aug 4;37(4). doi: 10.1042/BSR20170710. Print 2017 Aug 31.
The present study investigated the effects of microRNA-374 (miR-374) on human squamous cell carcinoma (SCC) cell proliferation, migration, invasion, and apoptosis through P53 signaling pathway by targeting growth arrest and DNA-damage-inducible protein 45 α (Gadd45a). Skin samples were collected from patients with skin SCC and normal skin samples. Expression of miR-374, Gadd45a, P53, P73, P16, c-myc, bcl-2, Bax, caspase-3, and caspase-9 were detected using quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. A431 and SCL-1 cells were divided into blank, negative control (NC), miR-374 mimics, miR374 inhibitors, siRNA-Gadd45a, and miR-374 inhibitors + siRNA-Gadd45a groups. Their proliferation, migration, invasion, cell cycle, and apoptosis were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, scratch test, Transwell assay, and flow cytometry. SCC skin tissues exhibited decreased expression of miR-374, P73, P16, Bax caspase-3 and caspase-9, and increased levels of Gadd45a, P53, c-myc, and Bcl-2 compared with the normal skin tissues. The miR-374 inhibitors group exhibited decreased expression of miR-374, P73, P16, Bax caspase-3 and caspase-9, and increased expression of Gadd45a, P53, c-myc, and Bcl-2, enhanced cell proliferation, migration, and invasion, and reduced apoptosis compared with the blank and NC groups; the miR-374 mimics group followed opposite trends. Compared with the blank and NC groups, the miR-374 inhibitors + siRNA-Gadd45a group showed decreased miR-374 level; the siRNA-Gadd45a group showed elevated levels of P73, P16, Bax, caspase-3 and caspase-9, decreased levels of Gadd45a, P53, c-myc, and Bcl-2, reduced cell proliferation, migration, and invasion, and accelerated apoptosis. miR-374 induces apoptosis and inhibits proliferation, migration, and invasion of SCC cells through P53 signaling pathway by down-regulating Gadd45a.
本研究通过靶向生长停滞和 DNA 损伤诱导蛋白 45α（Gadd45a），探讨微小 RNA-374（miR-374）对人鳞状细胞癌（SCC）细胞增殖、迁移、侵袭和凋亡的影响及其对 P53 信号通路的影响。收集皮肤 SCC 患者和正常皮肤样本的皮肤样本。采用实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测 miR-374、Gadd45a、P53、P73、P16、c-myc、bcl-2、Bax、caspase-3 和 caspase-9 的表达。将 A431 和 SCL-1 细胞分为空白组、阴性对照组（NC）、miR-374 模拟物组、miR-374 抑制剂组、siRNA-Gadd45a 组和 miR-374 抑制剂+siRNA-Gadd45a 组。通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴盐（MTT）测定、划痕试验、Transwell 测定和流式细胞术评估细胞增殖、迁移、侵袭、细胞周期和凋亡。与正常皮肤组织相比，SCC 皮肤组织中 miR-374、P73、P16、Bax caspase-3 和 caspase-9 的表达降低，Gadd45a、P53、c-myc 和 Bcl-2 的水平升高。miR-374 抑制剂组的 miR-374、P73、P16、Bax caspase-3 和 caspase-9 的表达降低，Gadd45a、P53、c-myc 和 Bcl-2 的表达升高，细胞增殖、迁移和侵袭增强，细胞凋亡减少，与空白组和 NC 组相比；miR-374 模拟物组则呈现相反的趋势。与空白组和 NC 组相比，miR-374 抑制剂+siRNA-Gadd45a 组的 miR-374 水平降低；siRNA-Gadd45a 组的 P73、P16、Bax、caspase-3 和 caspase-9 水平升高，Gadd45a、P53、c-myc 和 Bcl-2 水平降低，细胞增殖、迁移和侵袭减少，细胞凋亡加速。miR-374 通过下调 Gadd45a 诱导 SCC 细胞凋亡，抑制细胞增殖、迁移和侵袭，激活 P53 信号通路。
Zotero 插件