Genetic Manipulation of Sirtuin 3 Causes Alterations of Key Metabolic Regulators in Melanoma.
Genetic Manipulation of Sirtuin 3 Causes Alterations of Key Metabolic Regulators in Melanoma.
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DOI：
10.3389/fonc.2021.676077
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发表时间：
2021
影响因子：
4.7
通讯作者：
Ahmad N
中科院分区：
文献类型：
--
作者：
Singh CK;George J;Chhabra G;Nihal M;Chang H;Ahmad N
线粒体sirtuin SIRT 3在细胞代谢和能量产生中起着关键作用，这使其成为包括黑色素瘤在内的癌症管理的明显靶标。先前，我们已经证明SIRT 3在人黑色素瘤中组成性上调，并且其抑制导致体外人黑色素瘤细胞和体内人黑色素瘤异种移植物的抗增殖作用。在这项研究中，我们在细胞培养和小鼠异种移植中采用敲低和过表达策略扩展了我们的数据，以进一步验证和建立SIRT 3在黑素细胞中的促增殖功能及其相关的潜在机制，特别是关注代谢调节。我们发现，短发夹RNA（shRNA）介导的SIRT 3基因敲低G361黑色素瘤细胞显示减少肿瘤的免疫缺陷Nu/Nu小鼠。相反，SIRT 3过表达的Hs 294 T黑色素瘤细胞显示肿瘤生长增加。这些作用与异种移植组织中增殖标记物（PCNA）、存活标记物（Survivin）和血管生成标记物（VEGF）的变化一致。此外，在体外培养系统中，我们使用PCR阵列确定SIRT 3敲低对SK-MEL-2细胞中葡萄糖代谢的影响。SIRT 3敲除导致参与葡萄糖（32个基因）和糖原（5个基因）代谢的调节和酶途径的总共37个基因的改变。使用独创性途径分析（IPA）对这些鉴定的基因的功能注释预测了响应SIRT 3敲低的细胞活力/增殖、肿瘤生长和活性氧（ROS）降低以及细胞凋亡增加的累积作用。此外，对SIRT 3调节基因的IPA基因网络分析揭示了除了几个黑素瘤相关基因之外，这些基因之间的相互作用。Sirtuin途径被鉴定为显示SIRT 3与代谢调控基因沿着与其他sirtuin相互作用的顶级经典途径之一。IPA分析还预测了HIF 1 α、PKM、KDM 8、PPARGC 1A、mTOR的抑制以及P53和CLPP的激活;这些基因参与了主要的癌症/黑色素瘤相关信号传导事件。总的来说，这些结果表明SIRT 3抑制影响细胞代谢，从而赋予针对黑素瘤的抗增殖反应。
The mitochondrial sirtuin SIRT3 plays key roles in cellular metabolism and energy production, which makes it an obvious target for the management of cancer, including melanoma. Previously, we have demonstrated that SIRT3 was constitutively upregulated in human melanoma and its inhibition resulted in anti-proliferative effects in vitro in human melanoma cells and in vivo in human melanoma xenografts. In this study, we expanded our data employing knockdown and overexpression strategies in cell culture and mouse xenografts to further validate and establish the pro-proliferative function of SIRT3 in melanocytic cells, and its associated potential mechanisms, especially focusing on the metabolic regulation. We found that short-hairpin RNA (shRNA) mediated SIRT3 knockdown in G361 melanoma cells showed diminished tumorigenesis in immunodeficient Nu/Nu mice. Conversely, SIRT3 overexpressing Hs294T melanoma cells showed increased tumor growth. These effects were consistent with changes in markers of proliferation (PCNA), survival (Survivin) and angiogenesis (VEGF) in xenografted tissues. Further, in in vitro culture system, we determined the effect of SIRT3 knockdown on glucose metabolism in SK-MEL-2 cells, using a PCR array. SIRT3 knockdown caused alterations in a total of 37 genes involved in the regulation and enzymatic pathways of glucose (32 genes) and glycogen (5 genes) metabolism. Functions annotation of these identified genes, using the ingenuity pathway analysis (IPA), predicted cumulative actions of decreased cell viability/proliferation, tumor growth and reactive oxygen species (ROS), and increased apoptosis in response to SIRT3 knockdown. Further, IPA gene network analysis of SIRT3 modulated genes revealed the interactions among these genes in addition to several melanoma-associated genes. Sirtuin pathway was identified as one of the top canonical pathways showing the interaction of SIRT3 with metabolic regulatory genes along with other sirtuins. IPA analysis also predicted the inhibition of HIF1α, PKM, KDM8, PPARGC1A, mTOR, and activation of P53 and CLPP; the genes involved in major cancer/melanoma-associated signaling events. Collectively, these results suggest that SIRT3 inhibition affects cellular metabolism, to impart an anti-proliferative response against melanoma.
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影响因子：
7.4
作者：
Ozden, Ozkan;Park, Seong-Hoon;Wagner, Brett A.;Song, Ha Yong;Zhu, Yueming;Vassilopoulos, Athanassios;Jung, Barbara;Buettner, Garry R.;Gius, David
通讯作者：
Gius, David
影响因子：
4.3
作者：
Loftus SK;Baxter LL;Cronin JC;Fufa TD;NISC Comparative Sequencing Program;Pavan WJ
通讯作者：
Pavan WJ
影响因子：
3.7
作者：
Li S;Banck M;Mujtaba S;Zhou MM;Sugrue MM;Walsh MJ
通讯作者：
Walsh MJ
影响因子：
9.7
作者：
Gutteridge RE;Singh CK;Ndiaye MA;Ahmad N
通讯作者：
Ahmad N
影响因子：
8.8
作者：
Allen, Eric L.;Ulanet, Danielle B.;Smolen, Gromoslaw A.
通讯作者：
Smolen, Gromoslaw A.