Differential knockdown of TGF-β ligands in a three-dimensional co-culture tumor- stromal interaction model of lung cancer.
Differential knockdown of TGF-β ligands in a three-dimensional co-culture tumor- stromal interaction model of lung cancer.
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DOI：
10.1186/1471-2407-14-580
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发表时间：
2014-08-09
期刊：
影响因子：
3.8
通讯作者：
Nagase T
中科院分区：
文献类型：
--
作者：
Horie M;Saito A;Noguchi S;Yamaguchi Y;Ohshima M;Morishita Y;Suzuki HI;Kohyama T;Nagase T
转化生长因子(TGF)-β通过调节癌细胞的增殖、侵袭和肿瘤微环境的重塑，在肿瘤的进展中起关键作用。癌相关成纤维细胞（CAFs）是基质细胞的主要类型，TGF-β信号被激活。在抑制TGF-β信号的策略中，靶向TGF-β配体的RNA干扰（RNAi）正成为一种有前景的工具。尽管临床前研究支持这种治疗策略的有效性，但其对体内肿瘤微环境的影响尚不清楚。此外，由于各种TGF-β配体异构体的敲低而产生的差异效应尚未被研究。因此，需要一个概括肿瘤-基质相互作用的实验模型来验证治疗剂。我们已经建立了肺癌的三维共培养模型，并证明了共培养成纤维细胞在促进癌细胞侵袭和分化中的功能作用。在这里，我们使用这个模型来研究TGF-β配体的敲低如何影响不同细胞类型的行为。我们制备了携带人工microrna的慢病毒载体，对人TGF-β1和TGF-β2进行了抑菌，并检测了它们在肺癌细胞和成纤维细胞中的作用。慢病毒载体能有效、选择性地抑制TGF-β配体的表达，对这些细胞表现出抗增殖作用。此外，TGF-β配体的敲低可减弱成纤维细胞介导的胶原凝胶收缩，并减少肺癌细胞在三维共培养中的侵袭。我们还观察到针对不同TGF-β亚型在肺癌细胞和成纤维细胞中的不同效果。我们的研究结果支持了rnai介导的TGF-β配体靶向可能通过其对癌症和基质细胞的作用对肺癌治疗有益的观点。这项研究进一步证明了这种三维共培养模型在检查治疗药物对肿瘤-基质相互作用的影响方面的有用性。本文的在线版本（doi:10.1186/1471-2407-14-580）包含补充材料，可供授权用户使用。
Transforming growth factor (TGF)-β plays a pivotal role in cancer progression through regulating cancer cell proliferation, invasion, and remodeling of the tumor microenvironment. Cancer-associated fibroblasts (CAFs) are the predominant type of stromal cell, in which TGF-β signaling is activated. Among the strategies for TGF-β signaling inhibition, RNA interference (RNAi) targeting of TGF-β ligands is emerging as a promising tool. Although preclinical studies support the efficacy of this therapeutic strategy, its effect on the tumor microenvironment in vivo remains unknown. In addition, differential effects due to knockdown of various TGF-β ligand isoforms have not been examined. Therefore, an experimental model that recapitulates tumor–stromal interaction is required for validation of therapeutic agents. We have previously established a three-dimensional co-culture model of lung cancer, and demonstrated the functional role of co-cultured fibroblasts in enhancing cancer cell invasion and differentiation. Here, we employed this model to examine how knockdown of TGF-β ligands affects the behavior of different cell types. We developed lentivirus vectors carrying artificial microRNAs against human TGF-β1 and TGF-β2, and tested their effects in lung cancer cells and fibroblasts. Lentiviral vectors potently and selectively suppressed the expression of TGF-β ligands, and showed anti-proliferative effects on these cells. Furthermore, knockdown of TGF-β ligands attenuated fibroblast-mediated collagen gel contraction, and diminished lung cancer cell invasion in three-dimensional co-culture. We also observed differential effects by targeting different TGF-β isoforms in lung cancer cells and fibroblasts. Our findings support the notion that RNAi-mediated targeting of TGF-β ligands may be beneficial for lung cancer treatment via its action on both cancer and stromal cells. This study further demonstrates the usefulness of this three-dimensional co-culture model to examine the effect of therapeutic agents on tumor–stromal interaction. The online version of this article (doi:10.1186/1471-2407-14-580) contains supplementary material, which is available to authorized users.
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