Department of Oncology, Binzhou People's Hospital, Binzhou, China.
Department of Dermatology, Binzhou People's Hospital, Binzhou, China.
Braz J Med Biol Res. 2019 Oct 10;52(10):e8385. doi: 10.1590/1414-431X20198385. eCollection 2019.
Malignant melanoma (MM) is one of the malignant tumors with highly metastatic and aggressive biological actions. Schizandrin A (SchA) is a bioactive lignin compound with strong anti-oxidant and anti-aging properties, which is stable at room temperature and is often stored in a cool dry place. Hence, we investigated the effects of SchA on MM cell line A375 and its underlying mechanism. A375 cells were used to construct an in vitro MM cell model. Cell viability, proliferation, apoptosis, and migration were detected by Cell Counting Kit-8, BrdU assay, flow cytometry, and transwell two-chamber assay, respectively. The cell cycle-related protein cyclin D1 and cell apoptotic proteins (Bcl-2, Bax, cleaved-caspase-3, and cleaved-caspase-9) were analyzed by western blot. Alteration of H19 expression was achieved by transfecting with pEX-H19. PI3K/AKT pathway was measured by detecting phosphorylation of PI3K and AKT. SchA significantly decreased cell viability in a dose-dependent manner. Furthermore, SchA inhibited cell proliferation and cyclin D1 expression. SchA increased cell apoptosis along with the up-regulation of pro-apoptotic proteins (cleaved-caspase-3, cleaved-caspase-9, and Bax) and the down-regulation of anti-apoptotic protein (Bcl-2). Besides, SchA decreased migration and down-regulated matrix metalloproteinases (MMP)-2 and MMP-9. SchA down-regulated lncRNA H19. Overexpression of H19 blockaded the inhibitory effects of SchA on A375 cells. SchA decreased the phosphorylation of PI3K and AKT while H19 overexpression promoted the phosphorylation of PI3K and AKT. SchA inhibited A375 cell growth, migration, and the PI3K/AKT pathway through down-regulating H19.
恶性黑色素瘤（MM）是一种具有高度转移性和侵袭性生物学行为的恶性肿瘤。五味子甲素（SchA）是一种具有强抗氧化和抗衰老特性的生物活性木质素化合物，在室温下稳定，通常存放在阴凉干燥的地方。因此，我们研究了 SchA 对 MM 细胞系 A375 及其潜在机制的影响。使用 A375 细胞构建体外 MM 细胞模型。通过细胞计数试剂盒-8、BrdU 检测、流式细胞术和 Transwell 双室测定分别检测细胞活力、增殖、凋亡和迁移。通过 Western blot 分析细胞周期相关蛋白 cyclin D1 和细胞凋亡蛋白（Bcl-2、Bax、cleaved-caspase-3 和 cleaved-caspase-9）。通过转染 pEX-H19 来改变 H19 的表达。通过检测 PI3K 和 AKT 的磷酸化来测量 PI3K/AKT 途径。SchA 呈剂量依赖性显著降低细胞活力。此外，SchA 抑制细胞增殖和 cyclin D1 表达。SchA 增加细胞凋亡，同时上调促凋亡蛋白（cleaved-caspase-3、cleaved-caspase-9 和 Bax），下调抗凋亡蛋白（Bcl-2）。此外，SchA 降低迁移并下调基质金属蛋白酶（MMP）-2 和 MMP-9。SchA 下调 lncRNA H19。H19 的过表达阻断了 SchA 对 A375 细胞的抑制作用。SchA 降低了 PI3K 和 AKT 的磷酸化，而过表达 H19 则促进了 PI3K 和 AKT 的磷酸化。SchA 通过下调 H19 抑制 A375 细胞生长、迁移和 PI3K/AKT 途径。
