Modulation of ATP8B1 Gene Expression in Colorectal Cancer Cells
Suggest its Role as a Tumor Suppressor

doi:10.2174/1568009622666220517092340

摘要

目的: 本研究旨在了解肿瘤抑制基因在结直肠癌发生和发展中的作用。背景: 散发性结直肠癌（CRC）通过不同的分子事件发展。18q染色体的缺失是腺瘤向癌进展过程中的一个显著事件。关于该事件的分子效应器的信息有限。早些时候，我们报道了ATP8B1是一种与大肠癌相关的新基因。ATP8B1属于P型ATP酶（P4 ATP酶）家族，其主要功能是促进磷脂的移位。摘要: 在本研究中，我们试图将位于染色体18q上的ATP8B1基因作为肿瘤抑制基因。方法:细胞培养、患者数据分析、稳定的ATP8B1过表达SW480细胞系的产生、病毒颗粒的制备、细胞转导、用CRISPR/Cas9产生稳定的ATP8B1敲除HT29细胞系、用shRNA产生稳定的ATP8B1基因敲除HT2 9细胞系、ATP8B1基因表达的定量、实时细胞增殖和迁移测定，细胞增殖测定、细胞迁移测定、蛋白质分离和western印迹、终点细胞活力测定、鞘磷脂摄取和流出、统计和计算分析。结果: 我们研究了本地患者数据，并确认肿瘤样本中ATP8B1的表达降低。CRC细胞系通过降低和提高ATP8B1水平进行工程化，这为研究其在癌症进展中的作用提供了工具。CRISPR/Cas9或shRNA强制降低ATP8B1表达与CRC细胞系HT29的生长和增殖增加有关。相反，ATP8B1的过度表达导致SW480细胞系的生长和增生减少。我们生成了一个ATP8B1下游的基因网络。此外，我们提供了ATP8B1水平调节对该网络的预测效果以及对脂肪酸代谢相关基因的可能影响。结论: 位于染色体18q上的肿瘤抑制基因（ATP8B1）可能参与大肠癌的进展。敲除该基因会导致细胞增殖率增加，细胞死亡减少，表明其作为肿瘤抑制因子的作用。增加该基因在结直肠癌细胞中的表达会减缓其生长并增加细胞死亡。这些证据表明ATP8B1作为肿瘤抑制基因的作用。

关键词: 结直肠癌，翻转酶，离子转运蛋白，肿瘤抑制基因，染色体18q，脂质转运

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DOI https://dx.doi.org/10.2174/1568009622666220517092340	Print ISSN 1568-0096
Publisher Name Bentham Science Publisher	Online ISSN 1873-5576

当代肿瘤药物靶点

大肠癌细胞中ATP8B1基因表达的调节提示其作为肿瘤抑制因子的作用

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图形摘要

当代肿瘤药物靶点

大肠癌细胞中ATP8B1基因表达的调节提示其作为肿瘤抑制因子的作用

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