An optical labeling-based proliferation assay system reveals the paracrine effect of interleukin-6 in breast cancer.
An optical labeling-based proliferation assay system reveals the paracrine effect of interleukin-6 in breast cancer.
复制标题
DOI：
10.1016/j.bbamcr.2014.10.004
复制
发表时间：
2015-01
期刊：
影响因子：
--
通讯作者：
Toi M
中科院分区：
文献类型：
--
作者：
Itou J;Tanaka S;Sato F;Akiyama R;Kawakami Y;Toi M
增殖分析是表征各种细胞类型的基本方法之一。在传统的细胞增殖分析中，不能随着时间的推移观察到相同的样本，也不能单独分析不同细胞群体中的特定组，例如癌细胞。为了克服这些局限性，我们建立了一种基于光学标记的细胞增殖检测系统，其荧光可以通过辐射不可逆地从绿色光转换为红色。在一次无毒的光转化事件后，每个细胞中的红色荧光强度因细胞分裂而减弱。由此，我们开发了一种简单的方法，通过监测红色荧光随时间的减少来量化细胞增殖。本研究表明，基于光学标记的细胞增殖实验是分析细胞增殖的一种可行的新方法，可以加深我们对细胞增殖机制调控机制的理解。我们使用这个新建立的系统来分析分泌型白介素6(IL-6)在癌细胞增殖中的作用，这一功能尚未得到充分的表征。IL-6基因敲除后，细胞增殖减少。然而，与表达IL-6的细胞共培养后，Kaeed标记的IL-6基因敲除细胞的增殖能力得到恢复。这些数据表明，在基底样乳腺癌细胞中，IL-6表现出旁分泌作用，正向调节细胞的增殖。因此，我们的结果表明癌细胞可以分泌信号分子，如IL-6，以支持其他癌细胞的增殖。
Proliferation analysis is one of the basic approaches to characterize various cell types. In conventional cell proliferation assays, the same sample cannot be observed over time, nor can a specific group within a heterogeneous population of cells, for example, cancerous cells, be analyzed separately. To overcome these limitations, we established an optical labeling-based proliferation assay system with the Kaede protein, whose fluorescence can be irreversibly photoconverted from green to red by irradiation. After a single non-toxic photoconversion event, the intensity of red fluorescence in each cell is reduced by cell division. From this, we developed a simple method to quantify cell proliferation by monitoring reduction of red fluorescence over time. This study shows that the optical labeling-based proliferation assay is a viable novel method to analyze cell proliferation, and could enhance our understanding of mechanisms regulating cell proliferation machinery. We used this newly established system to analyze the functions of secreted interleukin-6 (IL-6) in cancer cell proliferation, which had not been fully characterized. Reduction in proliferation was observed following IL-6 knockdown. However, after co-culturing with IL-6-expressing cells, the proliferation of Kaede-labeled IL-6-knockdown cells was restored. These data indicate that in basal-like breast cancer cells, IL-6 exhibits a paracrine effect to positively regulate cell proliferation. Our results thus demonstrate that cancer cells can secrete signaling molecules, such as IL-6, to support the proliferation of other cancer cells.
登录
查看更多内容
影响因子：
10.8
作者：
Kang, Bin;Austin, Lauren A.;El-Sayed, Mostafa A.
通讯作者：
El-Sayed, Mostafa A.
影响因子：
8
作者：
Douglas, AM;Goss, GA;Begley, CG
通讯作者：
Begley, CG
影响因子：
4.6
作者：
Itou, Junji;Kawakami, Hiroko;Kawakami, Yasuhiko
通讯作者：
Kawakami, Yasuhiko
影响因子：
16.6
作者：
通讯作者：
--
影响因子：
4.7
作者：
Moon, Hyun-Yi;Kim, Oc-Hee;Kim, Cheol-Hee
通讯作者：
Kim, Cheol-Hee