Zang Jie, Zuo Dongqing, Shogren Kristen L, Gustafson Carl T, Zhou Zifei, Thompson Michael A, Guo Ruiwei, Prakash Y S, Lu Lichun, Guo Wei, Maran Avudaiappan, Yaszemski Michael J
Department of Orthopedic Surgery, Mayo Clinic, Rochester, MN 55905, USA.
Musculoskeletal Tumor Center, Peking University People's Hospital, Beijing 100044, China.
Chin J Cancer Res. 2019 Feb;31(1):203-211. doi: 10.21147/j.issn.1000-9604.2019.01.15.
To examine the role of store-operated calcium entry (SOCE) and stromal interaction molecule 1 (STIM1) in survival and migration of osteosarcoma cells and investigate what blockade of store-operated Ca contributes to the regulation of osteosarcoma cells.
First, we examined the expression levels of STIM1 in osteosarcoma cell lines by Western analysis and in tissue specimens by immunohistochemistry. Second, we investigated the effect of SOCE and STIM1 on osteosarcoma cell viability using MTS assays and on cell proliferation using colony formation. Third, we investigated the role of SOCE and STIM1 in cell migration using wound healing assays and Boyden chamber assays. Finally, we studied the effect of SOCE on the nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) activity by luciferase assays.
STIM1 was overexpressed in osteosarcoma cell lines and tissue specimens and was associated with poor survival of osteosarcoma patients. Also, inhibition of SOCE and STIM1 decreased the cell viability and migration of osteosarcoma cells. Furthermore, our results showed that blockade of store-operated Ca channels involved down-regulation of NFATc1 in osteosarcoma cells.
STIM1 is essential for osteosarcoma cell functions, and STIM1 and Ca entry pathway could be further explored as molecular targets in the treatment of osteosarcoma.
研究储存性钙内流（SOCE）和基质相互作用分子1（STIM1）在骨肉瘤细胞存活和迁移中的作用，并探讨阻断储存性钙内流对骨肉瘤细胞调控的影响。
首先，我们通过蛋白质免疫印迹分析检测骨肉瘤细胞系中STIM1的表达水平，并通过免疫组织化学检测组织标本中的表达水平。其次，我们使用MTS法研究SOCE和STIM1对骨肉瘤细胞活力的影响，使用集落形成法研究对细胞增殖的影响。第三，我们使用伤口愈合试验和博伊登小室试验研究SOCE和STIM1在细胞迁移中的作用。最后，我们通过荧光素酶试验研究SOCE对活化T细胞核因子胞质1（NFATc1）活性的影响。
STIM1在骨肉瘤细胞系和组织标本中过表达，且与骨肉瘤患者的不良生存相关。此外，抑制SOCE和STIM1可降低骨肉瘤细胞的活力和迁移能力。此外，我们的结果表明，阻断储存性钙通道涉及骨肉瘤细胞中NFATc1的下调。
STIM1对骨肉瘤细胞功能至关重要，STIM1和钙内流途径可作为骨肉瘤治疗的分子靶点进一步探索。
