Research On Stemness Of Prostate Cancer Cells Based On Multi-omics And Molecular Mechanism Of Compound Traditional Chinese Medicine CFF-1 Inhibiting Prostate Cancer

Tumor heterogeneity is the reason that promotes tumor evolution.The genomic variation of cancer cells and cancer stem cells with self-renewal and differentiation is related to tumor heterogeneity.Normal prostate tissue undergoes cancerous transformation to form a prostate intraepithelial tumor,develops into an invasive adenoma,and finally into aggressive prostate cancer.Cancer stem cells play an important role in cancer evolution,and the immune cells in the tumor microenvironment of prostate cancer are also related to transfer.Therefore,researching the role of cancer cell stemness in prostate cancer occurrence and development is of great significance to be the treatment and research of prostate cancer.Besides,traditional Chinese medicine has been used more and more widely in cancer treatment.Our laboratory has cooperated with the Department of Urology of Jiangsu Provincial Hospital of Traditional Chinese Medicine to explore the molecular mechanism of compound Chinese medicine CFF-1(in-hospital preparation)inhibiting the growth of prostate cancer cells.To explore the mechanism of prostate cancer occurrence and development more comprehensively,this article is based on multi-omics data,from the dimensions of genome mutation and evolution,chromatin accessibility,three-dimensional genome structure,transcription factor regulation,gene expression level,and single-cell gene expression,and so on,to discuss the stemness of prostate cancer cells.And the effective components and effective target molecules of compound Chinese medicine CFF-1 in the treatment of prostate cancer were analyzed by bioinformatics.The results show that:1.There are a large number of gene mutations and copy number variations in the genome of solid prostate tumors,and there is intratumoral heterogeneity in prostate cancer;based on the genomic variation information,the evolution model of prostate cancer cells are constructed,showing the linear evolution model of prostate cancer,and one type of cell subgroup exists in all evolution nodes may have cell stemness.2.Prostate cancer samples with high clinical grades have higher cancer cell stemness,according to the results of the tumor stemness score(OCLR)and clinical grade(Gleason grade)analysis.The gene modules related to the stemness of prostate cancer were obtained with WGCNA;the statistics of the copy number variation information of the stemness-related genes show that the degree of copy number variation of the stemness-related genes in samples with high Gleason grade is also higher,indicating that higher Gleason grade in prostate samples has stronger cancer cell stemness.3.The further analysis combined with the transcriptome expression data found that the high content of CD8+ T cells and M1 type macrophages in the high Gleason grade prostate cancer cells and the content of plasma cells was low;the stemness and immune infiltration of prostate cancer cells were negatively related.4.Based on ATAC-seq analysis,it is found that in prostate cancer cells and normal cells,the signal of open chromatin of transcription factors regulating stemness-related genes are different,and the transcription factors that contribute the most cells are also various;these transcription factors may promote the occurrence of prostate cancer and are the key factors affecting the stemness and metastasis of prostate cancer cells.5.Three-dimensional genome sequencing analysis of prostate cancer showed that the topological assocaited domains(TAD)of the cancer sample genome were generally reduced,and 214 chromatin loops that were significantly different from normal prostate cells were found in the prostate cancer genome;according to single-cell transcriptome data,prostate cancer cells were found to be present a subset of cells similar to cancer stem cells.6.The three-dimensional genome sequencing and single-cell sequencing to research the key gene EZH2 in prostate cancer showed that the expression of EZH2 is up-regulated in prostate cancer cells;the region where the EZH2 is located in the prostate cancer cell genome can form a specific chromatin loop,which may be related to the high expression of EZH2 in prostate cancer.7.Through network pharmacology,it was found that ESR1 of the estrogen signaling pathway and MAPK14 of the MAPK signaling pathway can interact with a large number of small molecules of Chinese medicine(active ingredients)in CFF-1.The self-developed Do＿virtual＿screening program was used to screen to obtain small drug molecules with the smallest binding free energy,and the dynamic binding process of small drug molecules with ESR1 and MAPK14 proteins was further simulated by molecular dynamics.This research found that the stemness of prostate cancer cells is related to gene mutations,particular transcription factors,immune microenvironment,and so on,deepening the understanding of the role of cancer cell stemness in prostate cancer.The discovery of a specific chromatin loop related to the high expression of the EZH2 in prostate cancer provides structure evidence for the research of the molecular mechanism of the EZH2 in prostate cancer.In addition,the small molecules(active ingredients)of traditional Chinese medicine that inhibit prostate cancer and its target proteins have been excavated from the perspective of bioinformatics,providing necessary guidance for further research and development of the compound traditional Chinese medicine CFF-1 for the treatment of prostate cancer.It also provides the necessary theoretical basis for opening up traditional Chinese medicine to enter the field of cancer treatment.