ligustilide and Prostatic-Neoplasms

The goal of this research was to study the selective pro-apoptotic effect of ligustilide on prostate-cancer-associated fibroblast in the tumor microenvironment and the related molecular mechanisms. The effects of ligustilide on cancer-associated fibroblasts (CAFs) and normal fibroblasts (NFs) isolated from the prostate were determined by MTT assay. Flow cytometry and cellular immunofluorescence were used to detect the effects of ligustilide on the cell cycle and apoptosis. Western blotting was used to detect the expression of apoptosis-related proteins after the action of ligustilide on CAFs. In the investigation, ligustilide had a selective pro-apoptotic effect on prostate-CAFs. After ligustilide treatment, the proportion of CAFs in the G2-M phase of the cell cycle increased, and the expression of apoptosis-related proteins (p-P53, Bcl-2, Caspase9 and Cytochrome C) changed. Ligustilide blocks the CAF cell cycle and induces the apoptosis of CAFs.

Topics: 4-Butyrolactone; Animals; Apoptosis; Cancer-Associated Fibroblasts; Cell Cycle; Cell Proliferation; Heterografts; Humans; Male; Mice; Mice, Inbred C57BL; Prostatic Neoplasms; Toll-Like Receptor 4; Tumor Microenvironment

The purpose of this work was to study the effects and underlying molecular mechanisms of ligustilide on cancer-associated fibroblasts (CAFs). The effects of ligustilide on the growth of CAFs and splenocytes were detected by MTT assay, and flow cytometry was used to detect effects on T-cell proliferation. Western blotting was used to detect the expression levels of CAF-related proteins after ligustilide treatment. This study found that ligustilide had no effect on the growth of splenocytes but that it could change the immunosuppressive function of CAFs through the TLR4-NF-κB pathway and restore T-cell proliferation previously inhibited by the CAF supernatant. Thus, ligustilide is expected to be a candidate for new antitumor drugs.

Topics: 4-Butyrolactone; Apoptosis; Cancer-Associated Fibroblasts; Cell Proliferation; Humans; Male; Prostatic Neoplasms; Signal Transduction; Tumor Cells, Cultured

Cancer development has been linked to epigenetic modifications of cancer oncogenes and tumor suppressor genes; in advanced metastatic cancers, severe epigenetic modifications are present. We previously demonstrated that the progression of prostate tumors in TRAMP mice is associated with methylation silencing of the Nrf2 promoter and a reduced level of transcription of Nrf2 and Nrf2 target genes. Radix Angelicae Sinensis (RAS; Danggui) is a medicinal herb and health food supplement that has been widely used in Asia for centuries. Z-Ligustilide (Lig) is one of the bioactive components of RAS. We investigated the potential of Lig and RAS to restore Nrf2 gene expression through epigenetic modification in TRAMP C1 cells. Lig and RAS induced the mRNA and protein expression of endogenous Nrf2 and Nrf2 downstream target genes, such as HO-1, NQO1, and UGT1A1. Bisulfite genomic sequencing revealed that Lig and RAS treatment decreased the level of methylation of the first five CpGs of the Nrf2 promoter. A methylation DNA immunoprecipitation assay demonstrated that Lig and RAS significantly decreased the relative amount of methylated DNA in the Nrf2 gene promoter region. Lig and RAS also inhibited DNA methyltransferase activity in vitro. Collectively, these results suggest that Lig and RAS are able to demethylate the Nrf2 promoter CpGs, resulting in the re-expression of Nrf2 and Nrf2 target genes. Epigenetic modifications of genes, including Nrf2, may therefore contribute to the overall health benefits of RAS, including the anticancer effect of RAS and its bioactive component, Lig.

Topics: 4-Butyrolactone; Angelica sinensis; Animals; Antineoplastic Agents, Phytogenic; Cell Line, Tumor; Cell Survival; CpG Islands; DNA Methylation; Drugs, Chinese Herbal; Epigenesis, Genetic; Gene Expression Regulation, Neoplastic; Male; Mice; NF-E2-Related Factor 2; Promoter Regions, Genetic; Prostate; Prostatic Neoplasms