背景:在发展过程中,胚胎采用不同的策略来去除不需要的细胞。使用类似于一些胚胎细胞的程序,我们从AMPK抑制的外周血单核细胞(PBMC)中的淋巴细胞和单核细胞中产生了一种消除肿瘤的条件培养基(CM).方法:AMPK信号通过应用药物抑制,Dorsomorphin,并使用体外细胞培养物评估其条件培养基(CM)的治疗效果,离体乳腺癌组织,以及乳腺肿瘤和肿瘤诱导的骨溶解的小鼠模型。使用基于质谱的蛋白质组学评估调节机制,西方印迹,免疫沉淀,基因过表达,和RNA干扰。结果:虽然AMPK信号主要起抗肿瘤作用,我们矛盾地抑制了它,以建立诱导的抑瘤细胞及其消除肿瘤的CM。在乳腺癌小鼠模型中,AMPK抑制的淋巴细胞衍生的CM的应用减少了乳腺肿瘤加化疗药物,紫杉醇.它还可以防止荷瘤胫骨的骨丢失。此外,来自患者外周血的CM的应用减少了从同一患者分离的离体乳腺癌组织。值得注意的是,富含CM的蛋白质包括Moesin(MSN),烯醇化酶1(ENO1),和polyA结合蛋白1(PABPC1),在许多类型的癌症中被认为是致瘤性的。MSN和ENO1的肿瘤抑制作用至少部分由Metadherin(Mtdh)介导,这是已知的促进转移播种。结论:我们证明PBMC可用于产生肿瘤抑制性蛋白质组,和细胞外肿瘤抑制蛋白如MSN,ENO1和PABPC1由癌细胞内的肿瘤促进因子转化。结果支持开发自体血液疗法的可能性,其中通过抑制AMPK信号传导在工程PBMC衍生的CM中富集肿瘤抑制蛋白。
Background: During a developmental process, embryos employ varying tactics to remove unwanted cells. Using a procedure analogous to some of the embryonic cells, we generated a tumor-eliminating conditioned medium (CM) from AMPK-inhibited lymphocytes and monocytes in peripheral blood mononuclear cells (PBMCs). Methods: AMPK signaling was inhibited by the application of a pharmacological agent, Dorsomorphin, and the therapeutic effects of their conditioned medium (CM) were evaluated using in vitro cell cultures, ex vivo breast cancer tissues, and a mouse model of mammary tumors and tumor-induced osteolysis. The regulatory mechanism was evaluated using mass spectrometry-based proteomics, Western blotting, immunoprecipitation, gene overexpression, and RNA interference. Results: While AMPK signaling acted mostly anti-tumorigenic, we paradoxically inhibited it to build induced tumor-suppressing cells and their tumor-eliminating CM. In a mouse model of breast cancer, the application of AMPK-inhibited lymphocyte-derived CM reduced mammary tumors additively to a chemotherapeutic agent, Taxol. It also prevented bone loss in the tumor-bearing tibia. Furthermore, the application of CM from the patient-derived peripheral blood diminished ex vivo breast cancer tissues isolated from the same patients. Notably, proteins enriched in CM included Moesin (MSN), Enolase 1 (ENO1), and polyA-binding protein 1 (PABPC1), which are considered tumorigenic in many types of cancer. The tumor-suppressing actions of MSN and ENO1 were at least in part mediated by Metadherin (Mtdh), which is known to promote metastatic seeding. Conclusion: We demonstrated that PBMCs can be used to generate tumor-suppressive proteomes, and extracellular tumor-suppressing proteins such as MSN, ENO1, and PABPC1 are converted from tumor-promoting factors inside cancer cells. The results support the possibility of developing autologous blood-based therapy, in which tumor-suppressing proteins are enriched in engineered PBMC-derived CM by the inhibition of AMPK signaling.