Effects of CD133 Silencing on Survival and Migration of HT-29 Colorectal Cancer Cells

Iran J Immunol. 2019 Sep;16(3):246-257. doi: 10.22034/IJI.2019.80275.

Abstract

Background: Colorectal cancer (CRC) is attributed as one of the most common malignancies worldwide. CD133 molecule, as a pentaspan transmembrane glycoprotein, confers stem cell-related characteristics, including self-renewal and multi-directional differentiation capability. CD133 plays important roles in the progression of CRC by conferring apoptotic resistance and migration ability.

Objective: To investigate the anti-apoptotic and anti-angiogenic effect of CD-133 targeted siRNA in a colorectal cancer cell line.

Methods: In this study, CD133-targeted siRNA transfection was conducted into HT-29 cells. MTT assay was employed to evaluate the cytotoxic effects of transfection on the cells. Flow cytometry was used to evaluate the apoptosis rate. The mRNA expression of apoptosis and metastasis related genes were assessed by quantitative Real-Time PCR (qRT-PCR). Wound healing assay was used to assess the migration potency of the infected cells.

Results: Expression of CD133 was significantly downregulated after transfection of CD133-specific siRNA. Moreover, the rate of apoptosis was significantly increased after transfection. The migration potential of cells was diminished after transfection. siRNA delivery resulted in the modulation of expression of apoptosis and metastasis-related genes.

Conclusion: siRNA mediated targeting of CD133 could be considered as a promising approach to treat CRC through suppressing the cancerous behavior of tumor cells.

MeSH terms

  • AC133 Antigen / genetics
  • AC133 Antigen / metabolism*
  • Apoptosis
  • Cell Movement
  • Cell Proliferation
  • Cell Survival
  • Colonic Neoplasms / genetics
  • Colonic Neoplasms / metabolism*
  • Colonic Neoplasms / pathology
  • Colorectal Neoplasms / genetics
  • Colorectal Neoplasms / metabolism*
  • Colorectal Neoplasms / pathology
  • Gene Expression Regulation, Neoplastic
  • Gene Silencing
  • HT29 Cells
  • Humans
  • Molecular Targeted Therapy
  • Neoplasm Metastasis
  • Neovascularization, Pathologic
  • RNA, Small Interfering / genetics

Substances

  • AC133 Antigen
  • RNA, Small Interfering