Expression of sarco/endoplasmic reticular Ca(2+)-ATPase in human lens epithelial cells and cultured human lens epithelial B-3 cells.
Expression of sarco/endoplasmic reticular Ca(2+)-ATPase in human lens epithelial cells and cultured human lens epithelial B-3 cells.
复制标题
人晶状体上皮细胞和培养的人晶状体上皮B-3细胞中肉质/内质网状Ca(2)-ATP酶的表达。
DOI：
10.1076/ceyr.19.5.389.5289
复制
发表时间：
1999
影响因子：
2
通讯作者：
Paterson,CA
中科院分区：
文献类型：
--
作者：
Liu,L;Bian,L;Borchman,D;Paterson,CA
SERCA 2b（100 Kd）和SERCA 3（97 Kd）在非肌肉组织中共表达。这两种类型的SERCA在非肌肉组织如脑和血小板中的Ca 2+调节中起重要作用。本研究以人晶状体上皮细胞、人透镜上皮B-3细胞（HLE B-3细胞）和大鼠晶状体上皮细胞为研究对象，检测了这两种SERCA在人晶状体上皮细胞、人透镜上皮B-3细胞（HLE B-3细胞）和大鼠晶状体上皮细胞中的表达，为进一步研究SERCA在白内障形成中的作用奠定基础。采用逆转录聚合酶链反应（RT-PCR）方法，以SERCA 2b和SERCA 3为引物，检测SERCA 3和SERCA 2b mRNA。用SERCA 2b和SERCA 3抗体分别在人透镜上皮细胞、人透镜上皮细胞B-3细胞和大鼠透镜上皮细胞中扩增出270 bp和186 bp的SERCA 3和SERCA 2b蛋白片段。两个片段均为预期的SERCA 2b和SERCA 3产物。这两个片段在人透镜上皮细胞中的序列分别与已发表的部分SERCA 2 b和SERCA 3 mRNA序列具有100%和98%的同源性。两个PCR产物所翻译的氨基酸序列与GeneBank中已发表的SERCA 2b和SERCA 3序列同源性均为100%。结论Sarco/内质网Ca 2+ -ATPase（SERCAs）亚型2b和3 mRNA在人透镜上皮细胞、培养的人透镜上皮细胞系（HLE B-3）和大鼠透镜上皮细胞中均有表达。SERCA 2 b和SERCA 3可能在调节透镜上皮细胞钙稳态中起重要作用。
PURPOSESarco/Endoplasmic Reticulum Ca 2+ -ATPase (SER-CAs) isoforms, SERCA2b (100Kd) and SERCA3 (97Kd), are co-expressed in some non-muscle tissues. Both types of SERCAs play important roles in Ca 2+ regulation in non-muscle tissues such as brain and platelets. We tested whether these two SERCAs are present in human lens epithelial cells, human lens epithelial B-3 cells (HLE B-3 cells) and rat lens epithelial cells providing a basis for further studies the role of SERCAs in cataract formation.METHODSLens epithelial cells from rat lens, human lens and cultured human lens epithelial B-3 cell line (HLE B-3) were used to extract mRNA and membrane proteins. Reverse transcription polymerase chain reaction (RT-PCR) was employed to detect SERCA3 and SERCA2b mRNA using SERCA2b and SERCA3 primers. Western blotting was used to detect SERCA3 and SERCA2b proteins using anti-SERCA2b and 3 antibodies.RESULTSA 270 bp fragment was amplified with SERCA2b primers and a 186 bp fragment amplified with SERCA3 primer in human lens epithelial cells, human lens epithelial B-3 cells and rat lens epithelial cells. Both fragments are the expected SERCA2b and SERCA3 products. The sequences of these two fragments in human lens epithelial cells are 100% and 98% homologous to the published partial SERCA2b and SERCA3 mRNA sequence, respectively. The amino acid sequences translated from both PCR products are 100% homologous to the published SERCA2b and SERCA3 sequences in GeneBank. Western blotting confirmed that the expected 100 KDa and a 97 KDa proteins are recognized by anti-SERCA2b and anti-SERCA3 antibodies.CONCLUSIONSarco/Endoplasmic Reticulum Ca 2+ -ATPase (SERCAs) isoforms 2b and 3 mRNAs are expressed in human lens epithelial cells, the cultured human lens epithelial B-3 cell line (HLE B-3) and rat lens epithelial cells. SERCA2b and SERCA3 may play an important role in the regulation of calcium homeostasis in lens epithelial cells.