The oncogene KRAS promotes cancer cell dissemination by stabilizing spheroid formation via the MEK pathway.
The oncogene KRAS promotes cancer cell dissemination by stabilizing spheroid formation via the MEK pathway.
复制标题
DOI：
10.1186/s12885-018-4922-4
复制
发表时间：
2018-12-03
期刊：
影响因子：
3.8
通讯作者：
Fujii T
中科院分区：
文献类型：
--
作者：
Ogishima J;Taguchi A;Kawata A;Kawana K;Yoshida M;Yoshimatsu Y;Sato M;Nakamura H;Kawata Y;Nishijima A;Fujimoto A;Tomio K;Adachi K;Nagamatsu T;Oda K;Kiyono T;Osuga Y;Fujii T
腹膜转移是卵巢癌的一个重要预后因素。虽然稳定的球状体形成促进卵巢癌的癌细胞腹膜转移，但相关的癌基因尚不清楚。在这项研究中，我们评估了KRAS癌基因在卵巢癌细胞播散中的作用，重点是细胞在球体条件下的稳定性，以及球体转化后细胞内信号的调节。在本研究中，我们使用了ID 8（一种鼠卵巢癌细胞系）和ID 8-KRAS（一种致癌KRAS（G12 V）转导的ID 8细胞系）。进行球体形成（3D）培养和细胞增殖测定以评价这些细胞的生长特性。cDNA微阵列分析进行，以确定在漂浮条件下参与KRAS相关信号转导的基因。MEK抑制剂用于评估对癌症腹膜扩散的影响。单层（2D）培养物中的细胞活力和增殖在ID 8和ID 8-KRAS细胞之间没有差异。然而，3D培养中活的和增殖的ID 8-KRAS细胞的比例分别比ID 8高约2倍和5倍。在ID 8-KRAS细胞中球体形成增加。对从腹膜内注射癌细胞的小鼠获得的腹膜漂浮细胞的分析显示，使用ID 8-KRAS的增殖癌细胞的比例比使用ID 8细胞的增殖癌细胞的比例高约2倍。全面的cDNA微阵列分析显示，与细胞增殖、细胞周期检查点和调控相关的通路在3D培养的ID 8-KRAS细胞中特异性上调，并且仅在3D培养的ID 8-KRAS细胞中上调了一些由MEK-ERK通路部分调控的基因。此外，MEK抑制剂曲美替尼在ID 8-KRAS细胞的3D培养中抑制球状体形成，尽管曲美替尼不影响2D培养细胞增殖。最后，我们证明了曲美替尼在体内小鼠模型中显著改善了ID 8-KRAS肿瘤小鼠的预后。我们的数据表明，KRAS促进卵巢癌扩散的稳定球体的形成和MEK途径是重要的稳定球体的形成。通过MEK抑制剂破坏球体形成可能是癌症腹膜传播的治疗靶点。本文的在线版本（10.1186/s12885-018-4922-4）包含补充材料，可供授权用户使用。
Peritoneal dissemination is a critical prognostic factor in ovarian cancer. Although stabilized spheroid formation promotes cancer cell peritoneal dissemination in ovarian cancer, the associated oncogenes are unknown. In this study, we assessed the role of the KRAS oncogene in ovarian cancer cell dissemination, focusing on the stability of cells in spheroid condition, as well as the modulation of intracellular signaling following spheroid transformation. We used ID8, a murine ovarian cancer cell line, and ID8-KRAS, an oncogenic KRAS (G12 V)-transduced ID8 cell line in this study. Spheroid-forming (3D) culture and cell proliferation assays were performed to evaluate the growth characteristics of these cells. cDNA microarray analysis was performed to identify genes involved in KRAS-associated signal transduction in floating condition. A MEK inhibitor was used to evaluate the effect on cancer peritoneal dissemination. Cell viability and proliferation in monolayer (2D) cultures did not differ between ID8 and ID8-KRAS cells. However, the proportions of viable and proliferating ID8-KRAS cells in 3D culture were approximately 2-fold and 5-fold higher than that of ID8, respectively. Spheroid-formation was increased in ID8-KRAS cells. Analysis of peritoneal floating cells obtained from mice intra-peritoneally injected with cancer cells revealed that the proportion of proliferating cancer cells was approximately 2-fold higher with ID8-KRAS than with ID8 cells. Comprehensive cDNA microarray analysis revealed that pathways related to cell proliferation, and cell cycle checkpoint and regulation were upregulated specifically in ID8-KRAS cells in 3D culture, and that some genes partially regulated by the MEK-ERK pathway were upregulated only in ID8-KRAS cells in 3D culture. Furthermore, a MEK inhibitor, trametinib, suppressed spheroid formation in 3D culture of ID8-KRAS cells, although trametinib did not affect 2D-culture cell proliferation. Finally, we demonstrated that trametinib dramatically improved the prognosis for mice with ID8-KRAS tumors in an in vivo mouse model. Our data indicated that KRAS promoted ovarian cancer dissemination by stabilizing spheroid formation and that the MEK pathway is important for stabilized spheroid formation. Disruption of spheroid formation by a MEK inhibitor could be a therapeutic target for cancer peritoneal dissemination. The online version of this article (10.1186/s12885-018-4922-4) contains supplementary material, which is available to authorized users.
登录
查看更多内容
影响因子：
7.3
作者：
Ahmed, Ashour Ahmed;Etemadmoghadam, Dariush;Temple, Jillian;Lynch, Andy G.;Riad, Mohamed;Sharma, Raghwa;Stewart, Colin;Fereday, Sian;Caldas, Carlos;DeFazio, Anna;Bowtell, David;Brenton, James D.
通讯作者：
Brenton, James D.
影响因子：
14.9
作者：
Nogales-Cadenas R;Carmona-Saez P;Vazquez M;Vicente C;Yang X;Tirado F;Carazo JM;Pascual-Montano A
通讯作者：
Pascual-Montano A
影响因子：
64.8
作者：
Mazur, Pawel K.;Reynoird, Nicolas;Khatri, Purvesh;Jansen, Pascal W. T. C.;Wilkinson, Alex W.;Liu, Shichong;Barbash, Olena;Van Aller, Glenn S.;Huddleston, Michael;Dhanak, Dashyant;Tummino, Peter J.;Kruger, Ryan G.;Garcia, Benjamin A.;Butte, Atul J.;Vermeulen, Michiel;Sage, Julien;Gozani, Or
通讯作者：
Gozani, Or
影响因子：
8
作者：
通讯作者：
--
影响因子：
158.5
作者：
Burger, Robert A.;Brady, Mark F.;Liang, Sharon X.
通讯作者：
Liang, Sharon X.