Melanoma is one of the most aggressive, potentially fatal forms of skin cancer and has been shown to be associated with solar ultraviolet radiation‐dependent initiation and progression. Despite remarkable recent advances with targeted and immune therapeutics, lasting and recurrence‐free survival remain significant concerns. Therefore, additional novel mechanism‐based approaches are needed for effective melanoma management. The sirtuin SIRT6 appears to have a pro‐proliferative function in melanocytic cells. In this study, we determined the effects of genetic manipulation of SIRT6 in human melanoma cells, in vitro and in vivo. Our data demonstrated that CRISPR/Cas9‐mediated knockout (KO) of SIRT6 in A375 melanoma cells resulted in a significant (1) decrease in growth, viability and clonogenic survival and (2) induction of G1‐phase cell cycle arrest. Further, employing a RT2 Profiler PCR array containing 84 key transformation and tumorigenesis genes, we found that SIRT6 KO resulted in modulation of genes involved in angiogenesis, apoptosis, cellular senescence, epithelial‐to‐mesenchymal transition, hypoxia signaling and telomere maintenance. Finally, we found significantly decreased tumorigenicity of SIRT6 KO A375 cells in athymic nude mice. Our data provide strong evidence that SIRT6 promotes melanoma cell survival, both in vitro and in vivo, and could be exploited as a target for melanoma management.

中文翻译：

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CRISPR/Cas9 介导的 SIRT6 敲除在体外和体内人类黑色素瘤细胞中产生显着的抗增殖反应

黑色素瘤是最具侵袭性、可能致命的皮肤癌之一，已被证明与太阳紫外线辐射依赖性的发生和进展有关。尽管最近在靶向和免疫疗法方面取得了显着进展，但持久和无复发生存率仍然是重大问题。因此，有效的黑色素瘤管理需要额外的基于机制的新方法。Sirtuin SIRT6 似乎在黑素细胞中具有促增殖功能。在这项研究中，我们在体外和体内确定了 SIRT6 基因操作对人类黑色素瘤细胞的影响。我们的数据表明，A375 黑色素瘤细胞中 CRISPR/Cas9 介导的 SIRT6 敲除 (KO) 导致显着 (1) 生长、活力和克隆存活率的降低，以及 (2) G1 期细胞周期停滞的诱导。此外，使用包含 84 个关键转化和肿瘤发生基因的 RT2 Profiler PCR 阵列，我们发现 SIRT6 KO 导致参与血管生成、细胞凋亡、细胞衰老、上皮间质转化、缺氧信号传导和端粒维持的基因的调节。最后，我们发现 SIRT6 KO A375 细胞在无胸腺裸鼠中的致瘤性显着降低。我们的数据提供了强有力的证据，表明 SIRT6 在体外和体内促进黑色素瘤细胞存活，并可作为黑色素瘤管理的靶点。最后，我们发现 SIRT6 KO A375 细胞在无胸腺裸鼠中的致瘤性显着降低。我们的数据提供了强有力的证据，表明 SIRT6 在体外和体内促进黑色素瘤细胞存活，并可作为黑色素瘤管理的靶点。最后，我们发现 SIRT6 KO A375 细胞在无胸腺裸鼠中的致瘤性显着降低。我们的数据提供了强有力的证据，表明 SIRT6 在体外和体内促进黑色素瘤细胞存活，并可作为黑色素瘤管理的靶点。