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Preclinical analysis of the anti-tumor and anti-metastatic effects of Raf265 on colon cancer cells and CD26(+) cancer stem cells in colorectal carcinoma.

Chow AK, Cheng NS, Lam CS, Ng L, Wong SK, Wan TM, Man JH, Cheung AH, Yau TC, Poon JT, Law WL, Pang RW - Mol. Cancer (2015)

Bottom Line: Raf265 was found to be highly effective in inhibiting cell proliferation and tumor growth through the inhibition of the RAF/MEK/ERK signaling pathway.In addition, anti-migratory and invasive effect was found with Raf265 treatment in combination with 5FU by targeting on the CD26(+) cells.Finally, the anti-tumor and anti-metastatic effect of Raf265 in combination with 5FU was also demonstrated.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, LKS Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Hong Kong, Hong Kong. chowakm@hku.hk.

ABSTRACT

Background: In colorectal carcinoma (CRC), activation of the Raf/MEK/ERK signaling pathway is commonly observed. In addition, the commonly used 5FU-based chemotherapy in patients with metastatic CRC was found to enrich a subpopulation of CD26(+) cancer stem cells (CSCs). As activation of the Raf/MEK/ERK signaling pathway was also found in the CD26(+) CSCs and therefore, we hypothesized that an ATP-competitive pan-Raf inhibitor, Raf265, is effective in eliminating the cancer cells and the CD26(+) CSCs in CRC patients.

Methods: HT29 and HCT116 cells were treated with various concentrations of Raf265 to study the anti-proliferative and apoptotic effects of Raf265. Anti-tumor effect was also demonstrated using a xenograft model. Cells were also treated with Raf265 in combination with 5FU to demonstrate the anti-migratory and invasive effects by targeting on the CD26(+) CSCs and the anti-metastatic effect of the combined treatment was shown in an orthotopic CRC model.

Results: Raf265 was found to be highly effective in inhibiting cell proliferation and tumor growth through the inhibition of the RAF/MEK/ERK signaling pathway. In addition, anti-migratory and invasive effect was found with Raf265 treatment in combination with 5FU by targeting on the CD26(+) cells. Finally, the anti-tumor and anti-metastatic effect of Raf265 in combination with 5FU was also demonstrated.

Conclusions: This preclinical study demonstrates the anti-tumor and anti-metastatic activity of Raf265 in CRC, providing the basis for exploiting its potential use and combination therapy with 5FU in the clinical treatment of CRC.

No MeSH data available.


Related in: MedlinePlus

The anti-proliferative effect of Raf265 on HT29 and HCT116 cells. A. Cells were treated with Raf265 at 0–50 μM and MTT assay was performed. B. Cells were suspended in the solidified agarose at the indicated concentrations of Raf265. Representing images under a phase-contrast microscopy at 40× magnification and an amplified view at 400× magnification were shown at the upper panel. The number of colony formed was then counted and the bar chart presenting the average number of colony formed was shown at the lower panel.
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Fig1: The anti-proliferative effect of Raf265 on HT29 and HCT116 cells. A. Cells were treated with Raf265 at 0–50 μM and MTT assay was performed. B. Cells were suspended in the solidified agarose at the indicated concentrations of Raf265. Representing images under a phase-contrast microscopy at 40× magnification and an amplified view at 400× magnification were shown at the upper panel. The number of colony formed was then counted and the bar chart presenting the average number of colony formed was shown at the lower panel.

Mentions: The effect of Raf265 on cell proliferation was measured by the MTT cell proliferation assay and the soft agar colony formation assay. Treatment of Raf265 for 72 hours inhibited cell proliferation in a dose dependent manner with an IC50 of 2.08 μM and 1.83 μM in HT29 and HCT116 cells, respectively (Figure 1A). Dose-dependent reduction in the number and size of colony formed in soft agar was also observed (Figure 1B). When treated with 1 μM Raf265 for 3 weeks, the number of colony formed reduced from 38.6 ± 6.5 and 28.3 ± 3.5 to 1.67 ± 1.15 and 0.67 ± 0.58 colonies for HT29 and HCT116 cells, respectively.Figure 1


Preclinical analysis of the anti-tumor and anti-metastatic effects of Raf265 on colon cancer cells and CD26(+) cancer stem cells in colorectal carcinoma.

Chow AK, Cheng NS, Lam CS, Ng L, Wong SK, Wan TM, Man JH, Cheung AH, Yau TC, Poon JT, Law WL, Pang RW - Mol. Cancer (2015)

The anti-proliferative effect of Raf265 on HT29 and HCT116 cells. A. Cells were treated with Raf265 at 0–50 μM and MTT assay was performed. B. Cells were suspended in the solidified agarose at the indicated concentrations of Raf265. Representing images under a phase-contrast microscopy at 40× magnification and an amplified view at 400× magnification were shown at the upper panel. The number of colony formed was then counted and the bar chart presenting the average number of colony formed was shown at the lower panel.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4481075&req=5

Fig1: The anti-proliferative effect of Raf265 on HT29 and HCT116 cells. A. Cells were treated with Raf265 at 0–50 μM and MTT assay was performed. B. Cells were suspended in the solidified agarose at the indicated concentrations of Raf265. Representing images under a phase-contrast microscopy at 40× magnification and an amplified view at 400× magnification were shown at the upper panel. The number of colony formed was then counted and the bar chart presenting the average number of colony formed was shown at the lower panel.
Mentions: The effect of Raf265 on cell proliferation was measured by the MTT cell proliferation assay and the soft agar colony formation assay. Treatment of Raf265 for 72 hours inhibited cell proliferation in a dose dependent manner with an IC50 of 2.08 μM and 1.83 μM in HT29 and HCT116 cells, respectively (Figure 1A). Dose-dependent reduction in the number and size of colony formed in soft agar was also observed (Figure 1B). When treated with 1 μM Raf265 for 3 weeks, the number of colony formed reduced from 38.6 ± 6.5 and 28.3 ± 3.5 to 1.67 ± 1.15 and 0.67 ± 0.58 colonies for HT29 and HCT116 cells, respectively.Figure 1

Bottom Line: Raf265 was found to be highly effective in inhibiting cell proliferation and tumor growth through the inhibition of the RAF/MEK/ERK signaling pathway.In addition, anti-migratory and invasive effect was found with Raf265 treatment in combination with 5FU by targeting on the CD26(+) cells.Finally, the anti-tumor and anti-metastatic effect of Raf265 in combination with 5FU was also demonstrated.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, LKS Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Hong Kong, Hong Kong. chowakm@hku.hk.

ABSTRACT

Background: In colorectal carcinoma (CRC), activation of the Raf/MEK/ERK signaling pathway is commonly observed. In addition, the commonly used 5FU-based chemotherapy in patients with metastatic CRC was found to enrich a subpopulation of CD26(+) cancer stem cells (CSCs). As activation of the Raf/MEK/ERK signaling pathway was also found in the CD26(+) CSCs and therefore, we hypothesized that an ATP-competitive pan-Raf inhibitor, Raf265, is effective in eliminating the cancer cells and the CD26(+) CSCs in CRC patients.

Methods: HT29 and HCT116 cells were treated with various concentrations of Raf265 to study the anti-proliferative and apoptotic effects of Raf265. Anti-tumor effect was also demonstrated using a xenograft model. Cells were also treated with Raf265 in combination with 5FU to demonstrate the anti-migratory and invasive effects by targeting on the CD26(+) CSCs and the anti-metastatic effect of the combined treatment was shown in an orthotopic CRC model.

Results: Raf265 was found to be highly effective in inhibiting cell proliferation and tumor growth through the inhibition of the RAF/MEK/ERK signaling pathway. In addition, anti-migratory and invasive effect was found with Raf265 treatment in combination with 5FU by targeting on the CD26(+) cells. Finally, the anti-tumor and anti-metastatic effect of Raf265 in combination with 5FU was also demonstrated.

Conclusions: This preclinical study demonstrates the anti-tumor and anti-metastatic activity of Raf265 in CRC, providing the basis for exploiting its potential use and combination therapy with 5FU in the clinical treatment of CRC.

No MeSH data available.


Related in: MedlinePlus