【摘要】 目的 研究不同病变宫颈组织中Eag1表达情况以及缺氧对宫颈癌Hela细胞膜上Eag1钾离子通道表达和活性的影响。方法 采用免疫组化法检测不同宫颈组织和宫颈癌Hela细胞中Eag1蛋白的表达情况;采用RT-PCR法检测不同缺氧时间和缺氧浓度对Hela细胞中Eag1 mRNA表达的影响。采用全细胞膜片钳记录的方法，研究缺氧对Eag1钾离子通道活性的影响。结果 宫颈上皮内瘤变组织中Eag1蛋白的表达明显高于正常组，宫颈癌组织中Eag1的表达明显高于宫颈上皮内瘤变组;Eag1的表达随临床分期的增加逐渐增高，Eag1的表达与病理分级无明显相关性。正常培养的宫颈癌Hela细胞中即有Eag1表达;其表达随缺氧程度加深及缺氧时间的延长明显增强。Hela细胞中Eag1钾离子通道是电压依赖性的，具有延迟整流及不失活特性;轻度缺氧条件能使Hela Eag1钾离子通道电流强度增大、激活时间缩短。结论 Eag1钾离子通道蛋白有可能成为新的早期筛查和判断宫颈癌生物学行为的诊断指标。
【关键词】 宫颈癌;钾离子通道;Eag1
Expression of Eag1 K+ channel in cancer of the cervix and the regulation of hypoxia
LIN Shuang,LI Li,LIU Yuan,et al. Department of Obstreics and Gynecology,Daping Hospital,The Third Military Medical University,Chongqing 400042,China
[Abstract] Objective Immunohistochemistry was employed to detect the expression of Eag1 protein in different cervix uteri tissues and CACX Hela cells.Methods We determined the effect of different hypoxia time and hypoxic concentration on the expression of Eag1 mRNA in Hela cell using RT-PCR. Meanwhile, whole cellular patch clamp recording was employed to investigate the effect of hypoxia on activity of Eag1 K+ channel.Results The expression of Eag1 protein in cervical intraepithelial neoplasia (CIN) tissue was significantly higher than that in normal control, and the expression in CACX tissue was remarkably higher than that in CIN group. The expression of Eag1 ascended as the development of clinical stage of cervical cancer, but almost irrelevant to pathological grade. Results of immunofluorescence and RT-PCR suggested that the expression of Eag1 significantly increased along with the aggravation of hypoxic degree and prolong of hypoxia time. Whole cellular patch clamp recording showed that slight anoxia condition could augment cell membrane potential of Hela, increase the current intensity of Eag1 K+ channel and shorten the activation time.Conclusion Eag1 K+ channel protein is possible to become diadynamic criteria for new early screeni
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