RNA interference-mediated knockdown of SIRT1 and/or SIRT2 in melanoma: Identification of downstream targets by large-scale proteomics analysis.
RNA interference-mediated knockdown of SIRT1 and/or SIRT2 in melanoma: Identification of downstream targets by large-scale proteomics analysis.
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DOI：
10.1016/j.jprot.2017.09.002
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发表时间：
2018-01-06
影响因子：
3.3
通讯作者：
Ahmad N
中科院分区：
文献类型：
--
作者：
Wilking-Busch MJ;Ndiaye MA;Liu X;Ahmad N
黑色素瘤是所有皮肤癌中最臭名昭著和最致命的，现有的治疗方案尚未被证明能有效地管理这种肿瘤，特别是转移疾病。Sirtuin(SIRT)蛋白已被证明在黑色素瘤中差异表达。我们已经证明，SIRT1和SIRT2在黑色素瘤中过表达，抑制SIRT1在人黑色素瘤细胞中产生抗增殖反应。为了阐明SIRT 1和/或2在黑色素瘤中的作用，我们利用shRNA介导的RNA干扰在A375人黑色素瘤细胞中稳定地敲除了SIRT 1、2及其组合。我们发现SIRT1和SIRT1&2联合基因敲除导致黑色素瘤细胞增殖下降。此外，SIRT 1和/或2的敲除导致黑色素瘤细胞集落形成减少。为了探索SIRT 1和/或2的下游靶点，我们利用稳定系进行了无标记的纳米LC-MS/MS蛋白质组学分析。我们发现异常水平的蛋白质参与了许多重要的细胞过程，包括细胞骨架组织、核糖体活性、氧化应激反应和血管生成。这些发现提供了细胞系统因sirtuin抑制而发生变化的明确证据，并为未来的研究揭示了几个很好的候选者。
Melanoma is the most notorious and fatal of all skin cancers and the existing treatment options have not been proven to effectively manage this neoplasm, especially the metastatic disease. Sirtuin (SIRT) proteins have been shown to be differentially expressed in melanoma. We have shown that SIRTs 1 and 2 were overexpressed in melanoma and inhibition of SIRT1 imparts anti-proliferative responses in human melanoma cells. To elucidate the impact of SIRT 1 and/or 2 in melanoma, we created stable knockdowns of SIRTs 1, 2, and their combination using shRNA mediated RNA interference in A375 human melanoma cells. We found that SIRT1 and SIRT1&2 combination knockdown caused a decreased cellular proliferation in melanoma cells. Further, the knockdown of SIRT 1 and/or 2 resulted in a decreased colony formation in melanoma cells. To explore the downstream targets of SIRTs 1 and/or 2, we employed a label-free quantitative nano-LC-MS/MS proteomics analysis using the stable lines. We found aberrant levels of proteins involved in many vital cellular processes, including cytoskeletal organization, ribosomal activity, oxidative stress response, and angiogenesis. These findings provide clear evidence of cellular systems undergoing alterations in response to sirtuin inhibition, and have unveiled several excellent candidates for future study.
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DOI：
10.1016/s1387-2656(05)11004-7
发表时间：
2005-01-01
期刊：
BIOTECHNOLOGY ANNUAL REVIEW, VOL 11
影响因子：
--
作者：
Berridge, MV;Herst, PM;Tan, AS
通讯作者：
Tan, AS
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
--
作者：
Kim HB;Lee SH;Um JH;Oh WK;Kim DW;Kang CD;Kim SH
通讯作者：
Kim SH
Ahnak 通过调节 TGFβ/Smad 信号通路发挥肿瘤抑制因子的作用。
影响因子：
8
作者：
通讯作者：
--
影响因子：
21.3
作者：
Fukata, Y;Itoh, TJ;Kaibuchi, K
通讯作者：
Kaibuchi, K