The aggressive metastatic phenotype of the HT29 LM1, LM2, and LM3 cell lines is associated with the activation of the c-MET pathway.

<p>HT29 LM1, LM2, LM3 and parental cell line were cultured in normal medium for 24(first 4 lanes). In another experiment, cells were cultured in serum free medium for 24 h and stimulated with complete medium for 10 min (remaining 4 lanes). Protein expression profiles were analyzed by Western blot. β-actin was used as a loading control.</p

Expression of CD44 is significantly increased in the HT29 LM1, LM2, and LM3 cell lines.

<p>(<b>A</b>) HT29 LM1, LM2, LM3 and parental cell line were cultured in normal medium for 24 h. Protein expression profiles were analyzed by Western blot. β-actin was used as a loading control. (<b>B</b>) IHC analysis of CD44 and p-MET expression in HT29 LM1 and HT29 LM3 liver metastasis tissue sections. (<b>C</b>) Flow cytometry analysis of CD44 geometric mean fluorescence intensity in parental HT29 and HT29 LM3 cells.</p

CD44 independent c-MET activation in HT29 LM3 cells.

<p>(<b>A</b>) Analysis of CD44, p-MET, c-MET, p-AKT and AKT expression in parental HT29 and HT29 LM3 after HGF stimulation (50 ng/mL; 5 min) by Western blot. β-actin was used as a loading control. (<b>B</b>) HT29 LM3 cells were transfected with CD44 siRNA and stimulated with HGF (50 ng/mL; 5 min) 24 h after siRNA transfection. (<b>C</b>) CD44 expression in HT29 LM3 and parental HT29 liver metastasis tissue sections were analyzed by IHC. (<b>D</b>) HT29 LM3 cells were sorted for high and low CD44 expression. Cells were gated and sorted to collect 10% of the cells with high CD44 expression and cells with low CD44 expression. (<b>E</b>) Analysis of CD44, p-MET, and c-MET expression in parental HT29, HT29 LM3 CD44−, and HT29 LM3 CD44+ cells. (<b>F</b>) Analysis of p-MET, c-MET, CD44, p-AKT, and AKT expression in parental HT29, HT29 LM3 CD44−, and HT29 LM3 CD44+ cells after HGF (50 ng/mL; 5 min) stimulation.</p

Delivery of RNA Nanoparticles into Colorectal Cancer Metastases Following Systemic Administration

The majority of deaths from all cancers, including colorectal cancer (CRC), is a result of tumor metastasis to distant organs. To date, an effective and safe system capable of exclusively targeting metastatic cancers that have spread to distant organs or lymph nodes does not exist. Here, we constructed multifunctional RNA nanoparticles, derived from the three-way junction (3WJ) of bacteriophage phi29 motor pRNA, to target metastatic cancer cells in a clinically relevant mouse model of CRC metastasis. The RNA nanoparticles demonstrated metastatic tumor homing without accumulation in normal organ tissues surrounding metastatic tumors. The RNA nanoparticles simultaneously targeted CRC cancer cells in major sites of metastasis, such as liver, lymph nodes, and lung. Our results demonstrate the therapeutic potential of these RNA nanoparticles as a delivery system for the treatment of CRC metastasis