The cAMP/PKA/CREB and TGFβ/SMAD4 Pathways Regulate Stemness and Metastatic Potential in Colorectal Cancer Cells
The cAMP/PKA/CREB and TGFβ/SMAD4 Pathways Regulate Stemness and Metastatic Potential in Colorectal Cancer Cells
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DOI：
10.1158/0008-5472.can-22-1369
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发表时间：
2022-11-15
期刊：
影响因子：
11.2
通讯作者：
Aoki, Masahiro
中科院分区：
文献类型：
--
作者：
Fujishita, Teruaki;Kojima, Yasushi;Aoki, Masahiro
转移是大多数癌症患者死亡的原因。然而，在大肠癌转移的机制仍然在很大程度上未知。在这里，我们研究了结直肠癌细胞如何获得转移潜力，使用一种新的结直肠癌小鼠模型，自发发展肝转移，通过引入Ctnnb 1，Kras，Trp 53和Smad 4（CKPS）基因的零星突变产生。蛋白质组学分析显示，与非转移模型的结直肠癌细胞相比，转移模型的结直肠癌干细胞标志物ALCAM（CD 166）和PROM 1（CD 133）的表达升高。使用源自CKPS模型的结肠直肠癌细胞（CKPS细胞）的脾至肝转移测定证明了ALCAM和PROM 1在引发转移中的功能重要性。在2D和球状体培养中使用CKPS细胞进行的遗传和药理学分析显示，ALCAM和PROM 1的表达分别由cAMP/PKA/CREB和TGFI 3/SMAD 4途径正向和负向调节。CKPS小鼠和结直肠癌患者的原发灶和转移灶中均表达磷酸化CREB，CKPS细胞中CREB的敲除降低了其球体形成和转移启动能力。CREB抑制剂处理增强了伊立替康抑制CKPS细胞肝转移的作用。这些结果揭示了ALCAM和PROM 1以及它们的上游调节因子cAMP/PKA/CREB和TGFI 3/SMAD 4通路在维持结直肠癌细胞的干细胞性和转移潜能中的重要作用，并表明CREB抑制可能是针对转移性结直肠癌的潜在治疗策略。这项研究确定了维持结直肠癌细胞的干性和转移潜力所必需的信号通路，并提出CREB作为转移性结直肠癌的治疗靶点。
Metastasis is responsible for the majority of deaths of patients with cancer. However, mechanisms governing metastasis in colorectal cancer remain largely unknown. Here we investigated how colorectal cancer cells acquire metastatic potential using a novel mouse model of colorectal cancer that spontaneously develops liver metastasis, gen-erated by introducing sporadic mutations of Ctnnb1, Kras, Trp53, and Smad4 (CKPS) genes. Proteomic analyses revealed elevated expres-sion of colorectal cancer stem cell markers ALCAM (CD166) and PROM1 (CD133) in colorectal cancer cells from the metastatic model compared with those from a nonmetastatic model. Spleen-to-liver metastasis assays using colorectal cancer cells derived from the CKPS model (CKPS cells) demonstrated the functional importance of ALCAM and PROM1 in initiating metastasis. Genetic and pharma-cologic analyses using CKPS cells in 2D and spheroid culture revealed that expression of ALCAM and PROM1 is regulated positively and negatively by the cAMP/PKA/CREB and TGFI3/SMAD4 pathways, respectively. Consistently, phospho-CREB was expressed in both primary and metastatic lesions of CKPS mice and patients with colorectal cancer, and knockout of CREB in CKPS cells reduced their spheroid-forming and metastasis-initiating abilities. Treatment with a CREB inhibitor potentiated the effect of irinotecan in suppres-sing liver metastasis by CKPS cells. These results reveal the essential roles of ALCAM and PROM1, as well as their upstream regulators, the cAMP/PKA/CREB and TGFI3/SMAD4 pathways, in maintaining the stemness and metastatic potential of colorectal cancer cells and indicate that CREB inhibition may be a potential therapeutic strategy against metastatic colorectal cancer.Significance: This study identifies signaling pathways essential for maintaining the stemness and metastatic potential of colorectal cancer cells and proposes CREB as a therapeutic target in metastatic colorectal cancer.