Experimental Research Of The Promotion Effect Of TAMs/CCL5 On Prostate Metastasis Via Activating Cancer Stem Cells And The Intervention By FuZhengYiLiu Formula

ObjectiveProstate cancer stem cells(PCSCs)play a critical role in prostate cancer progression and metastasis,which remains an obstacle for successful prostate cancer treatment.Tumor-associated macrophages(TAMs)are the most abundant immune cell population within the tumor microenvironment.Systematic investigation of the interaction and network signaling between PCSCs and TAMs may help in searching for the critical target to suppress PCSCs and metastasis.And the anti-tumor effect of FuZheng YiLiu formula(FZYL)in PCa stem cells self-renewal and metastasis was researched.Methods1.The effects of CCL5 on the proliferation of prostate cancer cells were investigated using CCK-8 method or colony formation assay.2.Wound healing assay and transwell assay were conducted to measure the migration and invasion ability changes of prostate cancer cells after exogenous interventions as indicated,respectively.3.Western blotting was used to measure the expression of protein.4.The self-renewal activity of prostate cancer stem cells were investigated using mammosphere formation assay and flow cytometry.5.The downstream response genes of CCL5 were investigated using QPCR.6.The relationship between CCL5 and downstream pathways was verified using plasmid transfection and siRNA interference.7.The correlation between CCL5 and pathological features of prostate cancer were investigated using tissue microarray analysis and bioinformatic analysis.8.The effect of TAMs on prostate cancer in vivo were investigated using Xenograft tumor animal model.Results1.TAMs-secreted CCL5 promotes the invasion and the PCSCs subpopulation of prostate cancer cells in vitro.2.STAT3 is the primary gene responsive for CCL5 stimulation in prostate cancer.3.CCL5 promotes prostate cancer invasion and PCSCs selfrenewal via activating the.CCR5/β-catenin/STAT3 pathway.4.CCL5 knockdown in TAMs suppresses prostate cancer growth,bone metastasis and PCSCs activity in vivo.5.CCL5 expression is positively correlated with advanced clinicopathological characteristics and poor prognosis of prostate cancer.6.The FZYL formula can inhibit the PCSCs subpopulation of prostate cancer cells in the co-culture system of TAMs and prostate cancer cells in vitro.7.The FZYL formula can inhibit the growth and bone metastasis of prostate cancer cells by blocking the effect of TAMs/CCL5 pathway in vivo.ConclusionTAMs/CCL5 could promote PCSCs self-renewal and prostate cancer metastasis via activating β-catenin/STAT3 signaling.Interestingly,it could be blocked by using FZYL.This study provides a novel rationale for developing TAMs/CCL5 as a potential molecular target for PCSCs elimination and metastatic prostate cancer prevention.