摘要
用半定量RT-PCR和流式细胞术,研究了白血病细胞系KG1a中P2X7受体在基因和蛋白水平的表达。用荧光分光光度计,测定了用激动剂三磷酸腺苷(ATP)和苯甲酰苯甲酸ATP(BzATP)刺激前后细胞内钙离子浓度的变化,证明P2X7受体的功能。结果表明:KG1a细胞表达P2X7受体,且在激动剂的刺激下能引起KG1a细胞通过P2X_7受体的胞外钙内流;去除胞外钙离子时,激动剂不能引起胞内钙浓度的升高;提示KG1a细胞表达P2X7受体的基因和功能蛋白,激活该受体引起胞外钙离子的内流。
The expression of ionotropic ATP-gated channel, P2X7 receptor, is studied at mRNA and protein level in human leukemia cell line KG1a by semi-quantitative RT-PCR and flow cytometry. In order to confirm whether the expressed P2X7 receptor has biological functions, cell Ca^2＋ is measured using the fluorescent dye Fura-2/AM. Intracellular free Ca^2＋ concentration （[Ca^2＋]i） increase in these cells induced by extracellular ATP as well as the more potent and specific agonist, 2′,3′-O-（4-Benzoylbenzoyl）-ATP （BzATP）, is determined by spectrophotometer. The results show that expression of P2X7 receptor is founded at both mRNA and protein level in KG1a cells. ATP and BzATP can induce extracellular Ca^2＋ influx and increase in [Ca^2＋]i through P2X7 channel in KG1a cells. Furthermore, ATP and BzATP cannot cause [Ca^2＋]i to increase in the absence of extracellular Ca^2＋ in KG1a cells. These results suggest that P2X7 receptor is functionally expressed in KG1a cells.
出处
《唐山师范学院学报》
2005年第5期33-36,53,共5页
Journal of Tangshan Normal University
基金
国家自然科学基金资助项目(30100072)
