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Knockdown by MSH2and EPCAMsiRNA Suppress Wnt/β-cateninPathway in HCT116 Cell Line
Wan Juhari WK1, Ahmad Amin Noordin KB2, Wan Abdul Rahman WF3, Zakaria AD4, Wan Muhamad Mokhter WMM5, Zilfalil BA6.
Small interfering RNA (siRNA) has the potential as a therapeutic approach against selective pathways in colorectal cancer(CRC). EPCAM, a transmembrane glycoprotein mediating cell adhesion, was known to be involved in suppressing Wnt/β-cateninpathway, an important pathway for tumour progression in colon cancer cells. EPCAMdeletions caused a transcriptional read-through that may silence its neighbouring gene, MSH2. This study aimed to investigate the effect of co-siRNA targeted genes, MSH2and EPCAM, in colon cancer cell line, HCT116, and their effect in regulating the Wnt/β-cateninpathway. Methods: Pre-designed siRNA of MSH2and EPCAMwere transfected into HCT116 cells. The cells were divided into six group of treatments: untreated cell group,cells treated with negative control siRNA, MSH2-siRNA treated cells, EPCAM-siRNA treated cells, cells treated with both EPCAMand MSH2-siRNA, and cells treated with transfection reagent (mock control). The mRNA and protein expression following the individual and combined siRNA treatments were assessed by quantitative polymerase chain reaction and Western blot. Results: The mRNA and protein expression levels of MSH2, EPCAMand β-cateninwere reduced in the individual MSH2and EPCAM-siRNA treated samples as compared to the untreated sample. Further reduction of mRNA and protein expressions for MSH2, EPCAMand β-cateninwere identified in combined siRNA treatments. Conclusion: Reduction of β-cateninexpression by simultaneous silencing of MSH2and EPCAMsuggested that these genes may play a role in supressing the Wnt/β-cateninpathway in cancer cells
Affiliation:
- Hospital Universiti Sains Malaysia, Malaysia
- Hospital Universiti Sains Malaysia, Malaysia
- Hospital Universiti Sains Malaysia, Malaysia
- Hospital Universiti Sains Malaysia, Malaysia
- Hospital Universiti Sains Malaysia, Malaysia
- Hospital Universiti Sains Malaysia, Malaysia
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