Qian Chengwei, Dai Yongzheng, Xu Xiaoli, Jiang Yong
Preventive Dentistry Laboratory, College and Hospital of Stomatology, Anhui Medical University.
Preventive Dentistry Laboratory, College and Hospital of Stomatology, Anhui Medical University
Ann Clin Lab Sci. 2019 Sep;49(4):457-467.
This study aims to explore the regulatory mechanism of hypoxia-inducible factor HIF-1α on Kv3.4. Oral squamous cell carcinoma (OSCC) cell lines SCC3 and CAL27 were used in this study. Western blotting and qRT-PCR methods were used to detect Kv3.4 expression levels in OSCC and their adjacent tissues. The expression changes of Kv3.4 and HIF-1α in a hypoxic environment were detected in cell lines. The stable OSCC cell lines with knockouts of HIF-1α and Kv3.4 were constructed. Transwell and CCK-8 assays were used to detect changes in the invasion, migration and proliferation ability after transfection. Chromatin immunoprecipitation and luciferase reporter gene assays were used to determine the regulatory and binding sites of HIF-1α on Kv3.4. The expression level of Kv3.4 in oral cancer tissue was higher than normal oral epithelium's regular value. The expression level of HIF-1α and Kv3.4 increased under hypoxia. Knocking out HIF-1α and Kv3.4 could reduce the invasion, migration and proliferation of cells. A down regulation of HIF-1α will reduce the Kv3.4 expression level. Overexpressing Kv3.4 after knocking down HIF-1α partially restored the proliferation and invasion of cell lines. Therefore, HIF-1α regulates the invasion, migration and proliferation of oral cancer cells by regulating Kv3.4 expression.
本研究旨在探讨缺氧诱导因子HIF-1α对Kv3.4的调控机制。本研究采用口腔鳞状细胞癌（OSCC）细胞系SCC3和CAL27。运用蛋白质免疫印迹法和qRT-PCR方法检测OSCC及其癌旁组织中Kv3.4的表达水平。检测细胞系在缺氧环境下Kv3.4和HIF-1α的表达变化。构建HIF-1α和Kv3.4基因敲除的稳定OSCC细胞系。采用Transwell实验和CCK-8实验检测转染后侵袭、迁移和增殖能力的变化。运用染色质免疫沉淀实验和荧光素酶报告基因实验确定HIF-1α对Kv3.4的调控及结合位点。口腔癌组织中Kv3.4的表达水平高于正常口腔上皮的正常水平。在缺氧条件下，HIF-1α和Kv3.4的表达水平升高。敲除HIF-1α和Kv3.4可降低细胞的侵袭、迁移和增殖能力。下调HIF-1α会降低Kv3.4的表达水平。敲低HIF-1α后过表达Kv3.4可部分恢复细胞系的增殖和侵袭能力。因此，HIF-1α通过调控Kv3.4的表达来调节口腔癌细胞的侵袭、迁移和增殖。
