Department of Clinical Biochemistry, Copenhagen University Hospital, Rigshospitalet, 2100 Copenhagen, Denmark.
Department of Clinical Biochemistry, Research Institute Nordstjernevej 38, Rigshospitalet, 2600 Glostrup, Denmark.
Cells. 2020 Oct 22;9(11):2341. doi: 10.3390/cells9112341.
Multiple myeloma is a malignant expansion of plasma cells and aggressively affects bone health. We show that P2X7 receptor altered myeloma growth, which affects primary bone cells in vitro. Expression on six human myeloma cell lines confirmed the heterogeneity associated with P2X7 receptor. Pharmacology with 2'(3')-O-(4-benzoylbenzoyl) adenosine 5'-triphosphate (BzATP) as agonist showed dose-dependent membranal pores on RPMI-8226 ( = 0.0027) and blockade with P2X7 receptor antagonists. Ca influx with increasing doses of BzATP ( = 0.0040) was also inhibited with antagonists. Chronic P2X7 receptor activation reduced RPMI-8226 viability ( = 0.0208). No apoptosis or RPMI-8226 death was observed by annexin V/propidium iodide (PI) labeling and caspase-3 cleavage, respectively. However, bromodeoxyuridine (BrdU) labelling showed an accumulation of RPMI-8226 in the S phase of cell cycle progression (61.5%, = 0.0114) with significant decline in G0/G1 (5.2%, = 0.0086) and G2/M (23.5%, = 0.0015) phases. As myeloma pathology depends on a positive and proximal interaction with bone, we show that P2X7 receptor on RPMI-8226 inhibited the myeloma-induced suppression on mineralization ( = 0.0286) and reversed the excessive osteoclastic resorption. Our results demonstrate a view of how myeloma cell growth is halted by P2X7 receptor and the consequences on myeloma-osteoblast and myeloma-osteoclast interaction in vitro.
多发性骨髓瘤是浆细胞的恶性扩张，严重影响骨骼健康。我们表明，P2X7 受体改变了骨髓瘤的生长，这会影响体外的原发性骨细胞。六种人类骨髓瘤细胞系的表达证实了与 P2X7 受体相关的异质性。用 2'（3'）-O-（4-苯甲酰基苯甲酰基）腺苷 5'-三磷酸（BzATP）作为激动剂的药理学研究表明，RPMI-8226 上存在剂量依赖性的膜孔（=0.0027），并且 P2X7 受体拮抗剂可阻断。随着 BzATP 剂量的增加，Ca 内流（=0.0040）也被拮抗剂抑制。慢性 P2X7 受体激活降低了 RPMI-8226 的活力（=0.0208）。用 Annexin V/碘化丙啶（PI）标记和 caspase-3 切割分别观察到没有细胞凋亡或 RPMI-8226 死亡。然而，溴脱氧尿苷（BrdU）标记显示 RPMI-8226 在细胞周期进展的 S 期积累（61.5%，=0.0114），G0/G1 期（5.2%，=0.0086）和 G2/M 期（23.5%，=0.0015）显著下降。由于骨髓瘤病理学依赖于与骨骼的积极和近端相互作用，我们表明 RPMI-8226 上的 P2X7 受体抑制了骨髓瘤诱导的矿化抑制（=0.0286）并逆转了过度的破骨细胞吸收。我们的研究结果表明，P2X7 受体如何阻止骨髓瘤细胞的生长，以及这种作用对体外骨髓瘤-成骨细胞和骨髓瘤-破骨细胞相互作用的影响。
