Department of Oral Medicine, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Biochemistry & Molecular Cell Biology, Shanghai Key Laboratory of Tumor Microenvironment and Inflammation, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Cell Mol Med. 2019 Jun;23(6):4326-4337. doi: 10.1111/jcmm.14324. Epub 2019 Apr 11.
Oral lichen planus (OLP) is considered a precancerous lesion with no known cure. Recent studies reported that abnormal regulation of apoptosis was involved in the pathogenesis of OLP. Next generation sequencing was used to screen the candidate microRNAs and genes in biopsies from patients with OLP and healthy mucosa. Human oral keratinocytes were transfected into the related oligonucleotides of miR-27b-3p/cyclophilin D and their control groups. Apoptosis was detected by TdT-mediated dUTP nick end labelling and flow cytometry. The levels of mRNA and protein were detected by quantitative PCR, Western blots, and enzyme-linked immunosorbent assays, respectively. Luciferase assays were performed to detect the luciferase activities of miR-27b-3p and cyclophilin D. Here, we showed that basal epithelium apoptosis was reduced and the miR-27b-3p levels were decreased in clinical OLP samples. We also found that down-regulation of miR-27b-3p inhibited epithelial keratinocyte apoptosis by up-regulating cyclophilin D expression. Moreover, cyclophilin D increased the protein stability of Bcl2 through direct binding, and Bcl2 suppressed caspase9/3 activation and cytochrome C release. Taken together, these data showed that miR-27b-3p regulated keratinocyte apoptosis through cyclophilin D/Bcl2 signalling, suggesting the miR-27b-3p regulated the pathogenesis of OLP.
口腔扁平苔藓(OLP)被认为是一种癌前病变,目前尚无已知的治愈方法。最近的研究报告称,细胞凋亡的异常调节参与了 OLP 的发病机制。下一代测序技术被用于筛选 OLP 患者和健康黏膜活检组织中的候选 microRNAs 和基因。将相关的 miR-27b-3p/环孢素 D 和它们的对照组的寡核苷酸转染到人口腔角质形成细胞中。通过 TdT 介导的 dUTP 缺口末端标记和流式细胞术检测细胞凋亡。通过定量 PCR、Western blot 和酶联免疫吸附测定分别检测 mRNA 和蛋白水平。通过荧光素酶测定检测 miR-27b-3p 和环孢素 D 的荧光素酶活性。在这里,我们表明,在临床 OLP 样本中,基底上皮细胞凋亡减少,miR-27b-3p 水平降低。我们还发现,下调 miR-27b-3p 通过上调 cyclophilin D 的表达抑制上皮角质形成细胞凋亡。此外,环孢素 D 通过直接结合增加 Bcl2 的蛋白稳定性,Bcl2 抑制 caspase9/3 激活和细胞色素 C 释放。总之,这些数据表明,miR-27b-3p 通过 cyclophilin D/Bcl2 信号通路调节角质形成细胞凋亡,提示 miR-27b-3p 调节 OLP 的发病机制。