TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells.
TALENs-directed knockout of the full-length transcription factor Nrf1α that represses malignant behaviour of human hepatocellular carcinoma (HepG2) cells.
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TALENs 定向敲除全长转录因子 Nrf1 α，抑制人肝细胞癌细胞 (HepG2) 的恶性行为
DOI：
10.1038/srep23775
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发表时间：
2016-04-11
影响因子：
4.6
通讯作者：
Zhang Y
中科院分区：
文献类型：
--
作者：
Ren Y;Qiu L;Lü F;Ru X;Li S;Xiang Y;Yu S;Zhang Y
全长Nrf 1 α被加工成不同的同种型，它们共同调节维持细胞稳态和器官完整性所必需的基因，小鼠中Nrf 1的肝脏特异性缺失导致自发性肝癌。在此，我们报道了在低分化高转移性肝细胞癌中，人组成型Nrf 1 α而不是较小的Nrf 1 β/γ表达减弱或消失。因此，Nrf 1 α在肿瘤的生理病理发生发展中具有重要作用，但其特异性的病理生物学功能尚不清楚。为了解决这一问题，在人肝细胞癌（HepG 2）细胞中产生了TALENs定向敲除Nrf 1 α，而不是Nrf 1 β/γ。在扫描电子显微镜下观察到Nrf 1 α−/−细胞被拉长，具有细长的纺锤形和细胞间扩大的间隙。与野生型对照相比，Nrf 1 α−/−细胞的侵袭和迁移能力显著增加沿着细胞周期G2-M期阻滞和S期减少，并伴有细胞凋亡抑制。尽管Nrf 1 α−/−细胞的软琼脂集落形成略有增加，但其功能丧失显著促进了裸鼠皮下癌异种移植物的肝转移不良生长。结合分子表达结果，我们推测Nrf 1 α（及其主要衍生物）是抑制癌细胞过程（如EMT）和恶性行为（如迁移）的基因调控机制的必需品。
The full-length Nrf1α is processed into distinct isoforms, which together regulate genes essential for maintaining cellular homeostasis and organ integrity, and liver-specific loss of Nrf1 in mice results in spontaneous hepatoma. Herein, we report that the human constitutive Nrf1α, rather than smaller Nrf1β/γ, expression is attenuated or abolished in the case of low-differentiated high-metastatic hepatocellular carcinomas. Therefore, Nrf1α is of importance in the physio-pathological origin and development, but its specific pathobiological function(s) remains elusive. To address this, TALENs-directed knockout of Nrf1α, but not Nrf1β/γ, is created in the human hepatocellular carcinoma (HepG2) cells. The resulting Nrf1α−/− cells are elongated, with slender spindle-shapes and enlarged gaps between cells observed under scanning electron microscope. When compared with wild-type controls, the invasive and migratory abilities of Nrf1α−/− cells are increased significantly, along with the cell-cycle G2-M arrest and S-phase reduction, as accompanied by suppressed apoptosis. Despite a modest increase in the soft-agar colony formation of Nrf1α−/− cells, its loss-of-function markedly promotes malgrowth of the subcutaneous carcinoma xenograft in nude mice with hepatic metastasis. Together with molecular expression results, we thus suppose requirement of Nrf1α (and major derivates) for gene regulatory mechanisms repressing cancer cell process (e.g. EMT) and malignant behaviour (e.g. migration).