Cucurbitacin B Induces Hypermethylation of Oncogenes in Breast Cancer Cells

Planta Med. 2019 Mar;85(5):370-378. doi: 10.1055/a-0791-1591. Epub 2018 Nov 21.

Abstract

Breast cancer is a complex disease driven by multiple factors including both genetic and epigenetic alterations. Recent studies revealed that abnormal gene expression induced by epigenetic changes including aberrant promoter methylation plays a critical role in human breast carcinogenesis. Cucurbitacin B has antiproliferative activity against various human breast cancer cells, but the molecular mechanism is not completely understood. In this study, we explore the influence of cucurbitacin B from Trichosanthes cucumerina on the methylation status at the promoter of oncogenes c-Myc, cyclin D1, and survivin in breast cancer cell lines. Growth inhibitory effect of cucurbitacin B on breast cancer cells was assessed by MTT assay and colony formation assay. Methylation status of genomic DNA was determined by methylation-specific PCR. Gene and protein expression levels of all genes studied were analyzed by real-time RT-PCR and western blot. The results indicated that cucurbitacin B could inhibit cell growth in breast cancer cells. The oncogene promoters are usually hypomethylated in cancer cells. Upon cucurbitacin B treatment, upregulation of DNMT1 and obvious heavy methylation in the promoters of c-Myc, cyclin D1, and survivin, which consequently downregulated the expression of all these oncogenes, were observed. Hence, cucurbitacin B proved to be a potential cancer therapeutic agent, in part by inducing hypermethylation and silences the oncogenic activation.

MeSH terms

  • Antineoplastic Agents, Phytogenic / chemistry
  • Antineoplastic Agents, Phytogenic / pharmacology*
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics
  • Cell Line, Tumor
  • Cyclin D1 / genetics
  • DNA (Cytosine-5-)-Methyltransferase 1 / genetics
  • DNA (Cytosine-5-)-Methyltransferase 1 / metabolism
  • DNA Methylation / drug effects
  • Female
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Promoter Regions, Genetic / genetics
  • Proto-Oncogene Proteins c-myc / genetics
  • Survivin / genetics
  • Trichosanthes / chemistry*
  • Triterpenes / chemistry
  • Triterpenes / pharmacology*

Substances

  • Antineoplastic Agents, Phytogenic
  • BIRC5 protein, human
  • CCND1 protein, human
  • MYC protein, human
  • Proto-Oncogene Proteins c-myc
  • Survivin
  • Triterpenes
  • cucurbitacin B
  • Cyclin D1
  • DNA (Cytosine-5-)-Methyltransferase 1
  • DNMT1 protein, human