Proteomic analysis of media from lung cancer cells reveals role of 14-3-3 proteins in cachexia.
Proteomic analysis of media from lung cancer cells reveals role of 14-3-3 proteins in cachexia.
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DOI：
10.3389/fphys.2015.00136
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发表时间：
2015
影响因子：
4
通讯作者：
Andrade FH
中科院分区：
文献类型：
--
作者：
McLean JB;Moylan JS;Horrell EM;Andrade FH
目的：在诊断时，60% 的肺癌患者出现恶病质，这是一种严重的消耗综合征，会增加发病率和死亡率。肿瘤分泌多种导致恶病质肌肉萎缩的因素，但尚未确定所有这些因素。我们使用 Orbitrap 电喷雾电离质谱法来鉴定刘易斯肺癌细胞 (LCM) 条件培养基中新型恶病质诱导候选物。结果：在三个生物学重复中确认了 10058 个蛋白质。 33 种被鉴定为分泌蛋白，包括 14-3-3 蛋白，它们是高度保守的接头蛋白，已知有 200 多个结合伙伴。我们通过蛋白质印迹证实了 LCM 中胞外 14-3-3 蛋白的存在，并发现 LCM 含有比用 C2C12 肌管条件化的培养基更少的 14-3-3 含量。使用中和抗体，我们耗尽了肌管培养基中的细胞外 14-3-3 蛋白，从而导致肌球蛋白含量减少。我们在分化的 C2C12 肌管中鉴定出了 14-3-3 蛋白 CD13 的拟议受体，并发现通过 Bestatin 抑制 CD13 也会导致肌球蛋白含量减少。结论：我们的新发现表明，细胞外 14-3-3 蛋白可能充当先前未识别的肌因子，并可能通过 CD13 发出信号以帮助维持肌肉质量。
Aims: At the time of diagnosis, 60% of lung cancer patients present with cachexia, a severe wasting syndrome that increases morbidity and mortality. Tumors secrete multiple factors that contribute to cachectic muscle wasting, and not all of these factors have been identified. We used Orbitrap electrospray ionization mass spectrometry to identify novel cachexia-inducing candidates in media conditioned with Lewis lung carcinoma cells (LCM). Results: One-hundred and 58 proteins were confirmed in three biological replicates. Thirty-three were identified as secreted proteins, including 14-3-3 proteins, which are highly conserved adaptor proteins known to have over 200 binding partners. We confirmed the presence of extracellular 14-3-3 proteins in LCM via western blot and discovered that LCM contained less 14-3-3 content than media conditioned with C2C12 myotubes. Using a neutralizing antibody, we depleted extracellular 14-3-3 proteins in myotube culture medium, which resulted in diminished myosin content. We identified the proposed receptor for 14-3-3 proteins, CD13, in differentiated C2C12 myotubes and found that inhibiting CD13 via Bestatin also resulted in diminished myosin content. Conclusions: Our novel findings show that extracellular 14-3-3 proteins may act as previously unidentified myokines and may signal via CD13 to help maintain muscle mass.
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影响因子：
7.3
作者：
Gardino, Alexandra K.;Yaffe, Michael B.
通讯作者：
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影响因子：
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通讯作者：
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影响因子：
3.8
作者：
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通讯作者：
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