基因敲除
细胞生长
流式细胞术
生物
细胞内
细胞
线粒体
细胞迁移
线粒体ROS
细胞凋亡
分子生物学
细胞生物学
生物化学
作者
Yunyi Huang,Zhipei Chen,Xuanyi Chen,Xiaohua Chen,Meng Xu
摘要
34-kDa translocase of the outer mitochondrial membrane (TOMM34) has been reported highly expressed in many cancers and is positively correlated to poorer prognosis. Our prior study showed TOMM34 is highly expressed in oral squamous cell carcinoma (OSCC) and is closely related to TNM classification and tumor size. TOMM34 is also associated with lymph node metastasis and poorer overall survival and disease-free survival in HPV-negative OSCC.We knocked down TOMM34 in OSCC cells (SCC15, HPV positive; Cal27, HPV negative) with siRNA and over-expressed with plasmids. The effects of TOMM34 on cell proliferation, migration and invasion abilities were detected by EdU assay, CCK-8 assay, wound-healing assay, and Transwell assay. We also detected the mitochondrial morphology and the intracellular Reactive Oxygen Species (ROS) level by fluorescence staining and flow cytometry. Finally, we monitored the protein levels of ERK pathway-related molecules.TOMM34 knockdown decreased the proliferation in SCC15 and Cal27, and weakened the migration and invasion abilities as well. Mitochondria became shorter, in the shape of dots or short rods, suggesting that mitochondrial damage occurred. Intracellular ROS levels increased significantly after knockdown TOMM34 and decreased after over-expressing TOMM34. The phosphorylation levels of ERK1/2 and MEK1/2 in SCC15 were significantly higher than in Cal27. Besides, the phosphorylation levels of ERK1/2 and MEK1/2 were inhibited in SCC15 after knockdown of TOMM34, but not in Cal27.TOMM34 promotes the cell proliferation, migration, and invasion of OSCC. In addition, TOMM34 participates in maintaining the mitochondrial shape and reducing the intracellular ROS level to protect cancer cells. Furthermore, TOMM34 increases the activity of ERK1/2 and MEK1/2 in HPV-positive OSCC cells but not in HPV-negative.
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