Journal: British Journal of Cancer
Article Title: Cell line and patient-derived xenograft models reveal elevated CDCP1 as a target in high-grade serous ovarian cancer
Figure Lengend Snippet: Monoclonal antibody targeting of CDCP1 impedes HGSC PDX growth. ( A ) H&E and anti-CDCP1 (brown) staining of three patient tumours and PDXs developed from these tumours. Magnification, × 40. CDCP1 is expressed by malignant cells, detected mainly in the membrane with cytosolic staining also apparent. Scale bar is 50 μ m. ( B – D ) Mice were injected with cell slurry from the PDX of patient 28 along with antibody 10D7 or isotype matched control IgG (100 μ g). 10D7 and IgG treatments continued weekly at 25 mg kg −1 per mouse ( n =4 mice per treatment). ( B ) Representative images, at the time of killing of mice at week 7, of the peritoneal cavity of mice treated with IgG (left) or 10D7 (right). ( C ) Graphical representation of the total number of tumours (left), the weight of the largest tumour (middle) and the weight of all tumours combined (right) of groups of mice treated with IgG or 10D7. Data represent mean and standard deviation of each group. ( D ) Western blot analysis of lysates from four randomly selected IgG- and 10D7-treated PDX tumours recovered from mice. Lysates were examined for CDCP1, p-Src-Y416 (pSrc), Src and GAPDH.
Article Snippet: Whole-cell lysates were collected using RIPA buffer (Sigma-Aldrich) with 1 × Complete protease inhibitor cocktail (Roche, Castle Hill, NSW, Australia), 2 mM sodium vanadate, and 10 mM sodium fluoride, and used in western blot analysis as described previously ( He et al , 2015 ) using mouse monoclonal anti-p53 antibody DO-7 (Sigma-Aldrich), rabbit anti-CDCP1 carboxyl terminal antibody #4115, rabbit anti-phospho-Src-Tyr416 (p-Src-Y416) antibody #2101, mouse anti-Src antibody #2110 (Cell Signaling Technology) or a mouse monoclonal anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody (Merck Millipore, Kilsyth, VIC, Australia).
Techniques: Staining, Injection, Standard Deviation, Western Blot