Hei Hongya, Gao Jianjun, Dong Jibin, Tao Jie, Tian Lulu, Pan Wanma, Wang Hongyu, Zhang Xuemei
Department of Pharmacology, School of Pharmacy, Fudan University, Shanghai 201203, China.
Department of Bone Metabolism, Institute of Radiation Medicine, Fudan University, Shanghai, 200032, China.
Mol Cells. 2016 Jul;39(7):530-5. doi: 10.14348/molcells.2016.0033. Epub 2016 Jun 21.
Large conductance calcium-activated potassium (BK) channels participate in many important physiological functions in excitable tissues such as neurons, cardiac and smooth muscles, whereas the knowledge of BK channels in bone tissues and osteoblasts remains elusive. To investigate the role of BK channels in osteoblasts, we used transcription activator-like effector nuclease (TALEN) to establish a BK knockout cell line on rat ROS17/2.8 osteoblast, and detected the proliferation and mineralization of the BK-knockout cells. Our study found that the BK-knockout cells significantly decreased the ability of proliferation and mineralization as osteoblasts, compared to the wild type cells. The overall expression of osteoblast differentiation marker genes in the BK-knockout cells was significantly lower than that in wild type osteoblast cells. The BK-knockout osteoblast cell line in our study displays a phenotype decrease in osteoblast function which can mimic the pathological state of osteoblast and thus provide a working cell line as a tool for study of osteoblast function and bone related diseases.
大电导钙激活钾（BK）通道参与神经元、心脏和平滑肌等可兴奋组织中的许多重要生理功能，而关于骨组织和成骨细胞中BK通道的知识仍然知之甚少。为了研究BK通道在成骨细胞中的作用，我们使用转录激活样效应核酸酶（TALEN）在大鼠ROS17/2.8成骨细胞上建立了BK基因敲除细胞系，并检测了BK基因敲除细胞的增殖和矿化情况。我们的研究发现，与野生型细胞相比，BK基因敲除细胞作为成骨细胞的增殖和矿化能力显著降低。BK基因敲除细胞中成骨细胞分化标志物基因的总体表达明显低于野生型成骨细胞。我们研究中的BK基因敲除成骨细胞系表现出成骨细胞功能下降的表型，这可以模拟成骨细胞的病理状态，从而提供一个工作细胞系作为研究成骨细胞功能和骨相关疾病的工具。
