Functional expression of voltage-gated sodium channels in primary cultures of human cervical cancer
Functional expression of voltage-gated sodium channels in primary cultures of human cervical cancer
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DOI：
10.1002/jcp.20871
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发表时间：
2007-02-01
影响因子：
5.6
通讯作者：
Gomora, Juan Carlos
中科院分区：
文献类型：
--
作者：
Diaz, Daniel;Delgadillo, Dulce Maria;Gomora, Juan Carlos
宫颈癌(CAC)是世界上第三大最常见的女性癌症死因，也是发展中国家女性的第一大死因。几种离子通道在癌症中表达上调，实际上钾通道已被认为是CAC的肿瘤标志物和治疗靶点。电压门控钠通道(VGSC)的活性参与了前列腺癌和乳腺癌细胞的增殖、运动和侵袭；然而，这类通道在CAC中的作用还没有被探索过。在本研究中，我们从分子和电生理水平鉴定了人宫颈癌活检组织原代培养的VGSC。利用全细胞膜片钳技术，我们分离和鉴定了电压门控Na+电流是所有被研究细胞内向电流的主要成分。钠电流具有动力学、电压依赖性、对河豚毒素(TTX)阻断的敏感性和对[Na+](O)的依赖性。通过用标准RT-PCR和特异性引物分析编码TTX敏感的Na+通道α亚基的mRNAs的表达，我们在CAC原代培养和活检组织的总RNA中检测到Na(V)1.2、Na(V)1.4、Na(V)1.6和Na(V)1.7转录本。扩增产物的限制性内切酶分析与相应基因的分子性质一致。值得注意的是，在正常宫颈活检组织中只检测到Na(V)1.4钠通道的转录本。结果首次显示了VGSC在人CAC原代培养中的功能性表达，并提示这些通道可能被认为是该类型癌症的潜在分子标志物。J.细胞。物理。2007年，210：469478。(C)2006年Wiley-Liss，Inc.
Cervical cancer (CaC) is the third most frequent cause of death from cancer among women in the world and the first in females of developing countries. Several ion channels are upregulated in cancer, actually potassium channels have been suggested as tumor markers and therapeutic targets for CaC. Voltage-gated sodium channels (VGSC) activity is involved in proliferation, motility, and invasion of prostate and breast cancer cells; however, the participation of this type of channels in CaC has not been explored. In the present study, we identified both at the molecular and electrophysiological level VGSC in primary cultures from human cervical carcinoma biopsies. With the whole cell patch clamp technique, we isolated and identified a voltage-gated Na+ current as the main component of the inward current in all investigated cells. Sodium current was characterized by its kinetics, voltage dependence, sensitivity to tetrodotoxin (TTX) block and dependence to [Na+](o). By analyzing the expression of mRNAs encoding TTX-sensitive Na+ channel alpha subunits with standard RT-PCR and specific primers, we detected Na(v)1.2, Na(v)1.4, Na(v)1.6, and Na(v)1.7 transcripts in total RNA obtained from primary cultures and biopsies of CaC. Restriction enzyme analysis of PCR products was consistent with the molecular nature of the corresponding genes. Notably, only transcripts for Na(v)1.4 sodium channels were detected in biopsies from normal cervix. The results show for the first time the functional expression of VGSC in primary cultures from human CaC, and suggest that these channels might be considered as potential molecular markers for this type of cancer. J. Cell. Physiol. 210: 469478, 2007. (c) 2006 Wiley-Liss, Inc.