School of Biological Sciences, Dublin Institute of Technology, Dublin, Ireland
School of Biological Sciences, Dublin Institute of Technology, Dublin, Ireland.
Anticancer Res. 2014 Jun;34(6):2851-7.
BACKGROUND/AIM: The claudin family of proteins are key constituents of tight junctions and the aberrant expression of these proteins can contribute to de-stabilisation of tight junctions and thus to loss of cell polarity and cohesion. Increased expression of claudin-1 and claudin-7 has been observed in pre-invasive cervical lesions and cervical carcinomas. The present study attempted to assess the effect of claudin-1 and claudin-7 overexpression on the HeLa cervical carcinoma cell line, in terms of cell proliferation/viability, permeability, invasion and migration.
HeLa cells were stably transfected with expression vectors containing the claudin-1 and claudin-7 genes to produce two separate stable cell lines expressing claudin-1 and claudin-7, respectively. The stable cell lines were examined with regard to their invasion and migration abilities, cell permeability and cell proliferation/viability and compared to non-claudin-1 or -7 transfected HeLa.
The present study found that claudin-1 and claudin-7 affected the migratory ability of HeLa cells, reducing their ability to migrate in a gap closure assay compared to non-claudin-transfected HeLa cells. Monolayers of claudin-1 and claudin-7 transfected cells also displayed an increased transepithelial electrical resistance indicating decreased permeability compared to non-claudin-transfected HeLa. The study found that claudin-1 or claudin-7 expression had no effect on the proliferation or viability of HeLa cells. Claudin-1 or -7 expression also did not affect the invasive ability of HeLa cells with both stable cells lines and non-claudin-transfected HeLa cells all showing low invasive ability.
The results of the present study indicate that claudin-1 and claudin-7 overexpression alone does not contribute to increased tumorigenesis in cervical carcinoma, instead claudin-1 and - 7 expression in HeLa cells contribute to reducing the migratory ability of cells and decrease their permeability.
背景/目的:紧密连接蛋白家族的蛋白是紧密连接的关键组成部分,这些蛋白的异常表达可能导致紧密连接的不稳定,从而导致细胞极性和细胞间黏附的丧失。在癌前宫颈病变和宫颈癌中观察到 claudin-1 和 claudin-7 的表达增加。本研究试图评估 claudin-1 和 claudin-7 过表达对 HeLa 宫颈癌细胞系的影响,包括细胞增殖/活力、通透性、侵袭和迁移。
用含有 claudin-1 和 claudin-7 基因的表达载体稳定转染 HeLa 细胞,分别产生表达 claudin-1 和 claudin-7 的两个独立稳定细胞系。用这些稳定细胞系检测其侵袭和迁移能力、细胞通透性以及细胞增殖/活力,并与非 claudin-1 或 -7 转染的 HeLa 细胞进行比较。
本研究发现,claudin-1 和 claudin-7 影响 HeLa 细胞的迁移能力,与非 claudin 转染的 HeLa 细胞相比,在缝隙闭合测定中降低了迁移能力。claudin-1 和 claudin-7 转染细胞的单层也显示出更高的跨上皮电阻,表明与非 claudin 转染的 HeLa 相比通透性降低。研究发现,claudin-1 或 claudin-7 的表达对 HeLa 细胞的增殖或活力没有影响。claudin-1 或 -7 的表达也不影响 HeLa 细胞的侵袭能力,两种稳定细胞系和非 claudin 转染的 HeLa 细胞的侵袭能力均较低。
本研究结果表明,claudin-1 和 claudin-7 的过表达本身不会导致宫颈癌的肿瘤发生增加,相反,claudin-1 和 -7 在 HeLa 细胞中的表达有助于降低细胞的迁移能力并降低其通透性。