Deregulation of miR-34a, miR-221 and miR-222 after HT29 cells treatment with Silibinin encapsulated in polymersomes as an anti-cancer stem cell agent

Abstract

Colorectal Cancer (CRC) has the most common malignant gastrointestinal cancer which representing about13% of all malignant tumor. CRC Cancer Stem Cell is the major reasons for recurrence of disease cause of solid tumor metastasis, relapse of cancer after treatment and drug resistance. Silibinin, an herbal extract from milk thistle plant, has been identified as a potential cancer medicine that can target the signaling pathway of CSCs and change their abilities. In our study, the results of CSC confirmation test such as specific surface CD markers and ability to form colonospheres was indicated the HT-29 cells as CSC-CRC. To increase the effectiveness of Silibinin, and also, to evaluate therapeutic intentions on HT-29 cancer stem-like cells, we encapsulated Silibinin in polymersome nanoparticle and validated the anti-proliferative and apoptotic activities of this new patent by MTT assay, AnnexinV/PI method, cell cycle analysis and DAPI staining. Furthermore, the efficacy of drug on Multicellular Tumor Spheroid (MCTS) and single cell suspension was showed that SPN had succeed to decrees the expression level of CSC CD markers compared with control group. Follow by using miRNAs as a novel and minus invasive expertise for prognostic, RT-qPCR confirmed that SPNs can repress oncogenic miRNAs such as miR-221 and miR-222. Silibinin encapsulated in Polymersome Nanoparticles (SPNs) can also enhance the expression of tumor suppressor miR-34a and some of its proapoptotic target genes such as P53, BAX, CASP9, CASP3, and CASP8. Our results suggested that SPNs can be recognized as a new stimulant factor to direct the HT-29 cancer cells toward apoptosis pathways thorough modify expression of some miRNAs and their apoptotic target genes directly and/or indirectly.