Molecular diversity of neuronal-type calcium channels identified in small cell lung carcinoma.
Molecular diversity of neuronal-type calcium channels identified in small cell lung carcinoma.
复制标题
小细胞肺癌中神经元型钙通道的分子多样性。
DOI：
10.1016/s0025-6196(12)61144-6
复制
发表时间：
1992
影响因子：
8.9
通讯作者：
Lennon,VA
中科院分区：
文献类型：
--
作者：
Oguro-Okano,M;Griesmann,GE;Wieben,ED;Slaymaker,SJ;Snutch,TP;Lennon,VA
使用聚合酶链反应（PCR），我们确定了两个不同类别的神经元型电压门控钙通道（VGCC）的RNA转录在一个原型小细胞肺癌（SCLC）细胞系，SCC-9。设计寡核苷酸引物以编码已知神经元VGCC类别的α1-亚基共有的氨基酸序列。对来自两个独立PCR产物的互补DNA（cDNA）克隆进行测序，发现其中一个对应于脑A类VGCC片段，该片段被预测编码小脑浦肯野细胞特征性的P型VGCC（对二氢吡啶和ω-芋螺毒素不敏感），但以前未在人类中鉴定。第二个PCR产物与神经元和神经内分泌细胞的D类VGCC片段相同（除了一个保守的核苷酸差异），该片段编码L型VGCC（对二氢吡啶敏感）。通过北方印迹分析，两种cDNA均与SCC-9中获得的信使RNA（mRNA）杂交; D类cDNA还与人大脑皮质mRNA杂交，但均不与骨骼肌细胞系TE 671的mRNA杂交。虽然没有鉴定出对应于B类VGCC（N型）的cDNA，但已知SCLC表达对ω-芋螺毒素敏感的VGCC，并且当与来自Lambert-Eaton肌无力综合征患者的血清IgG复合时，与125 I标记的ω-芋螺毒素共沉淀。可以想象，在SCLC中表达的神经元型VGCC的多个类别可能具有独特的和相关的抗原决定簇，这些抗原决定簇可能引起抗神经元自身免疫反应。这可以解释一系列副肿瘤性神经系统疾病，包括Lambert-Eaton综合征和亚急性小脑变性。
Using the polymerase chain reaction (PCR), we identified RNA transcripts for two distinct classes of neuronal-type voltage-gated Ca2+channels (VGCC) in a prototypic small cell lung carcinoma (SCLC) cell line, SCC-9. Oligonucleotide primers were designed to encode amino acid sequences common to α1-subunits of known neuronal VGCC classes. Sequencing of complementary DNA (cDNA) clones derived from two independent PCR products revealed that one corresponded to a brain class A VGCC fragment predicted to encode a P-type VGCC (insensitive to dihydropyridines and ω-conotoxin) characteristic of cerebellar Purkinje cells but not previously identified in humans. The second PCR product was identical (except for one conservative nucleotide difference) to a fragment of the class D VGCC of neurons and neuroendocrine cells, which encodes an L-type VGCC (sensitive to dihydropyridines). By Northern blot analyses, both cDNAs hybridized to messenger RNAs (mRNAs) obtained from SCC-9; class D hybridized additionally to human cerebral cortical mRNA, but neither hybridized to mRNA from the skeletal muscle cell line TE671. Although no cDNA corresponding to class B VGCC (N-type) was identified, SCLCs are known to express VGCC that are sensitive to ω-conotoxin and coprecipitate with125I-labeled-ω-conotoxin when complexed with serum IgG from patients with the Lambert-Eaton myasthenic syndrome. The multiple classes of neuronal-type VGCC expressed in SCLC could conceivably have both unique and related antigenic determinants that may give rise to antineuronal autoimmune responses. This would account for a spectrum of paraneoplastic neurologic disorders including the Lambert-Eaton syndrome and subacute cerebellar degeneration.