Department of Otolaryngology, Head and Neck Surgery, The Second Xiangya Hospital, Central South University, Changsha, China.
J Gene Med. 2019 Oct;21(10):e3118. doi: 10.1002/jgm.3118. Epub 2019 Sep 4.
The present study aimed to investigate the functions and regulation mechanism of the transmembrane protease, serine 3 (TMPRSS3), which plays an important role in sensorineural hearing loss.
House Ear Institute-Organ of Corti 1 (HEI-OC1) cells, comprising auditory-related cells, were used in the present study. An overexpression vector and small hairpin RNA target on TMPRSS3 were designed and transfected into HEI-OC1 cells. Circular RNA (circRNA) sequencing was conducted and expression profiles were obtained. The circular structure of circRNAs was validated with a polymerase chain reaction and Sanger sequencing using convergent and divergent primers.
Overexpression of TMPRSS3 increased cell viability, whereas suppression of TMPRSS3 increased the percentage of apoptotic cells and decreased cell viability, compared to the control group. circRNA sequencing provided expression profiles indicating that the overexpression of TMPRSS3 increased the expression level of 195 circRNAs. Results of GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) studies indicated that the circRNAs are focused on the RAS signaling pathway. The pathway, circ-Slc41a2 (chr10: 82744115|82767120), miR-182 and Akt, might comprise one of the key cascades of TMPRSS3.
TMPRSS3 is an important molecule in the regulation of cell viability and cell apoptosis of HEI-OC1 cells. Its functions are dependent on the circ-Slc41a2, miR-182 and Akt cascade.
本研究旨在探讨跨膜丝氨酸蛋白酶 3（TMPRSS3）的功能和调控机制，该蛋白在感音神经性听力损失中发挥重要作用。
本研究使用包含听觉相关细胞的耳研究所-耳蜗 1 型（HEI-OC1）细胞。设计了 TMPRSS3 的过表达载体和小发夹 RNA 靶点，并转染到 HEI-OC1 细胞中。进行环状 RNA（circRNA）测序并获得表达谱。使用聚合酶链反应和 Sanger 测序，使用聚合酶链反应和 Sanger 测序，使用收敛和发散引物验证 circRNA 的环状结构。
与对照组相比，TMPRSS3 的过表达增加了细胞活力，而 TMPRSS3 的抑制增加了凋亡细胞的百分比并降低了细胞活力。circRNA 测序提供了表达谱，表明 TMPRSS3 的过表达增加了 195 个 circRNA 的表达水平。GO（基因本体论）和 KEGG（京都基因与基因组百科全书）研究的结果表明，circRNAs 集中在 RAS 信号通路。circ-Slc41a2（chr10：82744115|82767120）、miR-182 和 Akt 通路可能构成 TMPRSS3 的关键级联反应之一。
TMPRSS3 是调节 HEI-OC1 细胞细胞活力和细胞凋亡的重要分子。其功能依赖于 circ-Slc41a2、miR-182 和 Akt 级联反应。
