背景:三阴性乳腺癌(TNBC)的治疗选择有限。中医药在TNBC的治疗中起着重要作用。草本植物对ScutellariabarbataD.Don和Scleromitriondiffusum(威尔德。)R.J.Wang(SH)因其抗肿瘤特性而在临床实践中常用。已被证明对肿瘤相关疾病有良好的治疗作用,但是潜在的分子机制尚未完全解释。
目的:通过生物信息学,验证了主要来自癌症相关成纤维细胞(CAF)的IL6,与预后不良有关。此外,细胞和动物实验证实SH抑制肿瘤增殖,迁移,和通过抑制IL6/NFκB途径在原位肿瘤模型中的生长。
方法:GEO,使用TCGA和HPA数据库分析CAFs和IL6的预后价值,然后检测IL6资源。在生物信息学之后,通过体外和体内实验验证了CAFs和CAFs衍生的IL6对TNBC的影响。细胞克隆形成试验,伤口愈合试验,用Transwell法检测体外对CAFs和CAFs来源的IL6的促进作用和对SH的抑制作用。使用小鼠中的TNBC模型来证明体内CAF和CAF衍生的IL6的促进和SH的抑制。通过检测独特的分子,通过蛋白质印迹法探索NF-κB的生物学途径。
结果:生物信息学分析显示,高比例的CAFs和升高的IL6水平与TNBC的不良预后显著相关。同时,IL6被证明主要来源于CAF。在生物信息学的指示之后,体外实验表明,CAFs和IL6均能增强MDA-MD-231细胞的克隆形成和迁移能力(231),此外,CAFs的促进与IL6水平有关。基于这些数据,CAFs来源的IL6增强与NF-κB信号通路的激活密切相关,而SH可以减少活化。最后,体内实验证实了CAFs/CAFs衍生的IL6/NF-κB的促进作用和SH抑制的功效。
结论:生物信息学数据表明,较高比例的CAFs和较高水平的CAFs衍生的IL6与TNBC较差的存活率显著相关。CAFs和CAFs衍生的IL6被证明在体外和体内都能促进TNBC的进展,其过程与NF-κB的激活密切相关。SH抑制了TNBC的进展,证明与CAFs/CAFs衍生的IL6/NF-κB密切相关。
BACKGROUND: The treatment options for triple-negative breast cancer (TNBC) are limited. Traditional Chinese Medicine (TCM) plays an important role in the treatment of TNBC. The herb pair Scutellaria barbata D.Don and Scleromitrion diffusum (Willd.) R.J.Wang (SH) is commonly used in clinical practice for its anti-tumor properties. It has been proven to have good therapeutic effects on tumor-related diseases, but the underlying molecular mechanisms are not yet fully explained.
OBJECTIVE: Through bioinformatics, it was validated that IL6, primarily derived from cancer-associated fibroblasts (CAFs), is associated with poor prognosis. Additionally, cell and animal experiments confirmed that SH inhibits tumor proliferation, migration, and growth in an orthotopic tumor model by suppressing the IL6/NF-κB pathway.
METHODS: GEO, TCGA and HPA databases were used to analyze the prognostic value of CAFs and IL6, then IL6 resource was detected. After the bioinformatics, the influence of CAFs and CAFs-derived IL6 on TNBC was verified by experiments both in vitro and in vivo. Cell clone formation assay, wound-Healing assay, and Transwell assay were used to detect the promotion of CAFs and CAFs-derived IL6 and the inhibition of SH in vitro. TNBC model in mice was used to prove the promotion of CAFs and CAFs-derived IL6 and the inhibition of SH in vivo. The biological pathway of NF-κB was explored by western blotting through detecting unique molecules.
RESULTS: Bioinformatics analysis revealed that higher proportion of CAFs and elevated level of IL6 were significantly associated with poor prognosis in TNBC. At the same time, IL6 was proved predominantly derived from CAFs. After the indication of bioinformatics, experiments in vitro demonstrated that both CAFs and IL6 could enhance the clone formation and migration ability of MDA-MD-231 cells (231), furthermore, the promotion of CAFs was related with the level of IL6. Based on these data, mechanism was detected that CAFs-derived IL6 enhancement was closely related to the activation of NF-κB signaling pathway, while the activation can be reduced by SH. In the end, the promotion of CAFs/CAFs-derived IL6/NF-κB and the efficacy of SH inhibition were both confirmed by experiments in vivo.
CONCLUSIONS: Bioinformatics data indicates that higher proportion of CAFs and higher level of CAFs-derived IL6 are significantly related to poorer survival of TNBC. CAFs and CAFs-derived IL6 were proved to promote the progression of TNBC both in vitro and in vivo, and the process of which was significantly related to the activation of NF-κB. SH inhibited the progress of TNBC, which was proved to be closely related to CAFs/CAFs-derived IL6/NF-κB.