沉默miR-222上调PUMA基因表达诱导高转移潜能腺样囊性癌细胞凋亡

Silencing miR-222 up-regulates PUMA expression and induces apoptosis in highly metastatic adenoid cystic carcinoma cells

目的:研究沉默miR-222靶向p53上调凋亡调控因子(PUMA)对高转移潜能腺样囊性癌细胞系SACC-LM的影响。方法:用As-miR-222和对照(As-miR-222 NC)分别转染SACC-LM细胞,并设立空白对照,进行Transwell、CCK-8和流式细胞术,观察miR-222沉默对SACC-LM细胞迁移、增殖、凋亡和周期分布的影响。通过实时荧光定量PCR (qRT-PCR)与蛋白印迹测定(Western Blot)证实沉默miR-222靶向PUMA表达。结果:转染As-miR-222可抑制SACC-LM细胞的迁移和增殖,增加细胞凋亡率,并使G0/G1期的细胞比例显著增加(P<0. 01)。qRT-PCR与蛋白印迹实验证实,miR-222的沉默通过上调PUMA表达参与上述行为的调节。结论:沉默miR-222通过上调PUMA抑制SACC-LM细胞的迁移、增殖,诱导凋亡。

Objective: To study the effect of silencing miR-222 targeting p53 up-regulation of apoptosis regulator (PUMA) on highy metastatic adenoid cystic carcinoma cell line SACC-LM. Methods: SACC-LM cells were transfected with As-miR- 222 and As-miR-222 NC respectively. The blank group was set up simultaneously. The migration was detected by Transwell assay. The proliferation was detected by CCK-8 assay. Apoptosis and cell cycle were examined by flow cytometry. The expressions of miR-222 and PUMA were analyzed by qRT-PCR and Western Blot. Results: Transfection of As-miR-222 inhibited the migration and proliferation of SACC-LM cells, increased the rate of apoptosis, and significantly increased the proportion of cells in G0/G1 phase (P<0. 01). MiR-222 was down-regulated significantly (P<0.05), while PUMA was up-regulated as confirmed by RT-PCR and Western Blot (P<0.01). Conclusions: Silencing miR-222 inhibits the migration and proliferation, indaces apoptosis of SACC-LM cells by up-regulating PUMA.