Mitochondrial calcium uniporter activity is dispensable for MDA-MB-231 breast carcinoma cell survival.
Mitochondrial calcium uniporter activity is dispensable for MDA-MB-231 breast carcinoma cell survival.
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DOI：
10.1371/journal.pone.0096866
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Anderson ME
中科院分区：
文献类型：
--
作者：
Hall DD;Wu Y;Domann FE;Spitz DR;Anderson ME
通过线粒体 Ca2+ 单向转运蛋白 (MCU) 的钙摄取被认为对于调节细胞信号事件、能量状态和生存至关重要。通过最近鉴定出编码 MCU 蛋白复合物亚基的基因，现在可以对单向转运蛋白进行功能解剖。癌细胞表现出与线粒体 Ca2+ 水平改变相关的线粒体功能障碍的许多方面，包括细胞凋亡抵抗、活性氧产生增加和氧化代谢减少。我们使用公开数据库确定乳腺癌患者的预后与 MCU Ca2+ 传导孔亚基表达增加和 MICU1 调节亚基表达减少呈负相关。因此，我们假设乳腺癌细胞可能对 MCU 通道操作敏感。我们使用广泛研究的 MDA-MB-231 乳腺癌细胞系来研究用特定 siRNA 和腺病毒过表达构建体破坏或增加线粒体 Ca2+ 摄取的激活是否会使这些细胞对治疗相关的应激敏感。 MDA-MB-231 细胞被发现含有功能性 MCU 通道，这些通道很容易对细胞刺激做出反应，并在营养撤退时引发强烈的 AMPK 磷酸化反应。令人惊讶的是，MCU 或 MICU1 的敲低不会影响活性氧的产生，也不会对暴露于辐射、化疗药物或营养剥夺的 MDA-MB-231 细胞的克隆细胞存活产生显着影响。野生型或显性失活突变体 MCU 的过度表达不会影响基础克隆效率或神经酰胺诱导的细胞杀伤。相比之下，MCU 或 MICU1 敲除后，非癌性乳腺上皮 HMEC 细胞的存活率降低。这些结果支持这样的结论，即 MDA-MB-231 乳腺癌细胞不依赖于 MCU 或 MICU1 活性来生存，与之前在宫颈癌、结肠癌和前列腺癌衍生细胞中的发现相反，并表明并非所有癌症都对针对线粒体 Ca2+ 摄取机制的治疗敏感。
Calcium uptake through the mitochondrial Ca2+ uniporter (MCU) is thought to be essential in regulating cellular signaling events, energy status, and survival. Functional dissection of the uniporter is now possible through the recent identification of the genes encoding for MCU protein complex subunits. Cancer cells exhibit many aspects of mitochondrial dysfunction associated with altered mitochondrial Ca2+ levels including resistance to apoptosis, increased reactive oxygen species production and decreased oxidative metabolism. We used a publically available database to determine that breast cancer patient outcomes negatively correlated with increased MCU Ca2+ conducting pore subunit expression and decreased MICU1 regulatory subunit expression. We hypothesized breast cancer cells may therefore be sensitive to MCU channel manipulation. We used the widely studied MDA-MB-231 breast cancer cell line to investigate whether disruption or increased activation of mitochondrial Ca2+ uptake with specific siRNAs and adenoviral overexpression constructs would sensitize these cells to therapy-related stress. MDA-MB-231 cells were found to contain functional MCU channels that readily respond to cellular stimulation and elicit robust AMPK phosphorylation responses to nutrient withdrawal. Surprisingly, knockdown of MCU or MICU1 did not affect reactive oxygen species production or cause significant effects on clonogenic cell survival of MDA-MB-231 cells exposed to irradiation, chemotherapeutic agents, or nutrient deprivation. Overexpression of wild type or a dominant negative mutant MCU did not affect basal cloning efficiency or ceramide-induced cell killing. In contrast, non-cancerous breast epithelial HMEC cells showed reduced survival after MCU or MICU1 knockdown. These results support the conclusion that MDA-MB-231 breast cancer cells do not rely on MCU or MICU1 activity for survival in contrast to previous findings in cells derived from cervical, colon, and prostate cancers and suggest that not all carcinomas will be sensitive to therapies targeting mitochondrial Ca2+ uptake mechanisms.
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影响因子：
64.5
作者：
Mallilankaraman K;Doonan P;Cárdenas C;Chandramoorthy HC;Müller M;Miller R;Hoffman NE;Gandhirajan RK;Molgó J;Birnbaum MJ;Rothberg BS;Mak DO;Foskett JK;Madesh M
通讯作者：
Madesh M
影响因子：
2.9
作者：
Glancy, Brian;Balaban, Robert S.
通讯作者：
Balaban, Robert S.
DOI：
10.1126/science.1242993
发表时间：
2013-12-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Sancak Y;Markhard AL;Kitami T;Kovács-Bogdán E;Kamer KJ;Udeshi ND;Carr SA;Chaudhuri D;Clapham DE;Li AA;Calvo SE;Goldberger O;Mootha VK
通讯作者：
Mootha VK
影响因子：
16.6
作者：
Qiu J;Tan YW;Hagenston AM;Martel MA;Kneisel N;Skehel PA;Wyllie DJ;Bading H;Hardingham GE
通讯作者：
Hardingham GE
内膜的四十千克蛋白是线粒体钙Uniporter。
影响因子：
64.8
作者：
De Stefani, Diego;Raffaello, Anna;Teardo, Enrico;Szabo, Ildiko;Rizzuto, Rosario
通讯作者：
Rizzuto, Rosario