Tissue Microarray Cytometry Reveals Positive Impact of Homeodomain Interacting Protein Kinase 2 in Colon Cancer Survival Irrespective of p53 Function

Stage-dependent survival of the colorectal cancer patient series used to build the TMA.

Analysis of an independent series of C-C chemokine receptors double labeled by immunofluorescence for p53 and HIPK2. This TMA contains 30 colorectal cancers, eight of which have matched normal adjacent tissue. A: Percentage of p53-positive cells in normal and tumor cores. B: Survival analysis of the p53+ group (tumors with more than five percent p53-positive cells) and the p53- group (tumors with less than five percent p53-positive cells). C: Expression of HIPK2 in normal and tumor cores. D: Correlation plot between the tumor/normal ratio of HIPK2 expression and the survival (in months) for the eight patients with matched tissues. E: An example of patients with an increased expression of HIPK2 (core images: overlay of DAPI, p53 green, and HIPK2 red channels) and of the cytometric quantification of the signal on the normal and matched tumor cores, respectively.

Comparison of LSM with automated LSC acquisition and analysis. A: Correlation of the percentage of p53-positive cells in tumor cores obtained by LSM (x axis) and LSC (y axis). Homemade TMA: black circles, TMA from US Biomax; gray squares, the border color of the scattegram displays. B: Examples of biparametric scatterplots of paired samples shown in Figure 6 (left) and the same scatterplots obtained by LSC analysis; each plot shows p53 (x axis) and HIPK2 (y axis) nuclear fluorescence intensities in tumors (green) and paired normal mucosa (red). C: Correlation plot between the medians of HIPK2 intensities obtained by LSM (x axis) and LSC (y axis) in the homemade TMA. D: Correlation plot between the tumor/normal ratio of HIPK2 expression calculated on the basis of the LSM (x axis) and LSC (y axis) acquisition and analyses, respectively.