Morphology, Motility, and Cytoskeletal Architecture of Breast Cancer Cells Depend on Keratin 19 and Substrate.
Morphology, Motility, and Cytoskeletal Architecture of Breast Cancer Cells Depend on Keratin 19 and Substrate.
复制标题
DOI：
10.1002/cyto.a.24011
复制
发表时间：
2020-11
期刊：
影响因子：
--
通讯作者：
Chung BM
中科院分区：
文献类型：
--
作者：
Lam VK;Sharma P;Nguyen T;Nehmetallah G;Raub CB;Chung BM
癌细胞通过伴随细胞形状调节的事件获得运动性，包括改变角蛋白的表达。然而，角蛋白在癌细胞结构改变中的作用还不清楚。因此，我们消除了MDA-MB-231细胞系的乳腺癌细胞中角蛋白19（K19）的表达，并发现缺乏K19的细胞在培养中变得更长，在KRT 19转导后，形态学向亲本表型逆转。此外，肌动蛋白应力纤维和局灶性粘连的数量显着减少KRT 19敲除（KO）细胞。然后使用数字全息显微镜（DHM）定量表征KRT 19 KO细胞的改变的形态，这不仅证实了KRT 19 KO细胞的表型变化，而且还确定了K19依赖性形态变化依赖于底物类型。一种新的单细胞分析的定量方法，从DHM，通过平均相位差图，促进K19基板对细胞形态的相互作用的评价。当铺在胶原基质上时，KRT 19 KO细胞较少伸长并且类似于亲本细胞。评估单细胞运动性进一步显示，虽然KRT 19 KO细胞在刚性表面上比亲本细胞移动得更快，但当细胞接种在胶原上时，运动性的这种增加被消除。总体而言，我们的研究表明，K19抑制细胞运动通过调节细胞的形状在一个底物依赖性的方式。因此，这项研究提供了一个潜在的基础，角蛋白的表达改变与细胞形状和运动的癌细胞的变化。
Cancer cells gain motility through events that accompany modulation of cell shape and include altered expression of keratins. However, the role of keratins in change of cancer cell architecture is not well understood. Therefore, we ablated the expression of keratin 19 (K19) in breast cancer cells of the MDA-MB-231 cell line and found that cells lacking K19 become more elongated in culture, with morphological reversion toward the parental phenotype upon transduction of KRT19. Also, the number of actin stress fibers and focal adhesions were significantly reduced in KRT19 knockout (KO) cells. The altered morphology of KRT19 KO cells was then characterized quantitatively using digital holographic microscopy (DHM), which not only confirmed the phenotypic change of KRT19 KO cells but also identified that the K19-dependent morphological change is dependent on the substrate type. A new quantitative method of single cell analysis from DHM, via average phase difference maps, facilitated evaluation of K19–substrate interactive effects on cell morphology. When plated on collagen substrate, KRT19 KO cells were less elongated and resembled parental cells. Assessing single cell motility further showed that while KRT19 KO cells moved faster than parental cells on a rigid surface, this increase in motility became abrogated when cells were plated on collagen. Overall, our study suggests that K19 inhibits cell motility by regulating cell shape in a substrate-dependent manner. Thus, this study provides a potential basis for the altered expression of keratins associated with change in cell shape and motility of cancer cells.
登录
查看更多内容
通过数字全息显微镜在体外对伤口愈合的无定量染色和连续多模式监测。
影响因子：
3.7
作者：
Bettenworth D;Lenz P;Krausewitz P;Brückner M;Ketelhut S;Domagk D;Kemper B
通讯作者：
Kemper B
影响因子：
3.3
作者：
Hookway C;Ding L;Davidson MW;Rappoport JZ;Danuser G;Gelfand VI
通讯作者：
Gelfand VI
影响因子：
64.5
作者：
Cheung KJ;Gabrielson E;Werb Z;Ewald AJ
通讯作者：
Ewald AJ
影响因子：
64.8
作者：
Kim, S;Wong, P;Coulombe, PA
通讯作者：
Coulombe, PA
影响因子：
3.3
作者：
Bordeleau F;Galarneau L;Gilbert S;Loranger A;Marceau N
通讯作者：
Marceau N