Luo Min, Sun Gang, Sun Jing-Wu
Department of Otorhinolaryngology Head and Neck Surgery, Affiliated Provincial Hospital of Medical University of Anhui, Hefei 230001, Anhui, China.
Department of Otorhinolaryngology Head and Neck Surgery, Chaohu Hospital Affiliated to Medical University of Anhui, Chaohu 238000, Anhui, China.
Auris Nasus Larynx. 2019 Aug;46(4):583-592. doi: 10.1016/j.anl.2018.10.020. Epub 2018 Nov 16.
To explore the effect of miR-196bon the biological features of human laryngeal squamous cell carcinoma (LSCC) through targeting PCDH-17.
miR-196b and PCDH-17 expressions were determined in tissues, and the targeting relation of miR-196b and PCDH-17 was verified through dual-luciferase reporter system. In vitro, Hep-2 cells were divided into the Control, miR-196b inhibitors, miR-NC, PCDH-17, and miR-196b mimics+PCDH-17 groups. The miR-196b and PCDH-17 expressions were determined by qRT-PCR or/and Western blot, and the biological features by MTT, Annexin V-FITC/PI, wound-healing and Transwell assays.
MiR-196b was found to be up-regulated, while PCDH-17 was down-regulated in a negative correlation in LSCC patients, which was related to histological grade and TNM stage. And low expression of miR-196b and high expression of PCDH-17 contributed to an increase in the 5-year-survival rate of LSCC patients. Besides, miR-196b directly targeted PCDH-17, while miR-196b inhibitors could up-regulate the PCDH-17 in Hep-2 cells. Moreover, miR-196b inhibitors and PCDH-17 curbed Hep-2 cell proliferation but facilitated the apoptosis, with decreases in cell invasion and migration. In addition, no statistical significance was found in cell proliferation, apoptosis, invasion and migration between Control group and miR-196b mimics+PCDH-17 group.
LSCC patients exhibited the up-regulated miR-196b and down-regulated PCDH-17, which are correlated with the major clinical features and prognosis. Inhibiting miR-196b may suppress proliferation, migration and invasion abilities, and promote apoptosis of Hep-2 cells via targeting PCDH-17.
通过靶向原钙黏蛋白17（PCDH-17）探讨微小RNA-196b（miR-196b）对人喉鳞状细胞癌（LSCC）生物学特性的影响。
检测组织中miR-196b和PCDH-17的表达，并通过双荧光素酶报告系统验证miR-196b与PCDH-17的靶向关系。体外实验中，将人喉癌细胞系Hep-2细胞分为对照组、miR-196b抑制剂组、阴性对照miR-NC组、PCDH-17组以及miR-196b模拟物+PCDH-17组。采用实时定量聚合酶链反应（qRT-PCR）或/和蛋白质免疫印迹法检测miR-196b和PCDH-17的表达，通过噻唑蓝（MTT）比色法、膜联蛋白V异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）双染法、划痕愈合实验及Transwell实验检测细胞生物学特性。
LSCC患者中miR-196b呈上调表达，PCDH-17呈下调表达，二者呈负相关，且与组织学分级和TNM分期相关。miR-196b低表达和PCDH-17高表达有助于提高LSCC患者五年生存率。此外，miR-196b直接靶向PCDH-17，miR-196b抑制剂可上调Hep-2细胞中PCDH-17的表达。而且，miR-196b抑制剂和PCDH-17均可抑制Hep-2细胞增殖，但促进细胞凋亡，并降低细胞侵袭和迁移能力。此外，对照组与miR-196b模拟物+PCDH-17组在细胞增殖、凋亡、侵袭及迁移方面无统计学差异。
LSCC患者中miR-196b上调，PCDH-17下调，二者与主要临床特征及预后相关。抑制miR-196b可能通过靶向PCDH-17抑制Hep-2细胞增殖、迁移和侵袭能力，并促进其凋亡。
