Hepatocellular carcinoma microvessel density quantitation with image analysis: Correlation with prognosis

Hepatocellular carcinoma (HCC) has a progression considered to be dependent on angiogenesis. Intratumoral microvessel density (MVD) has been associated with metastasis and recurrence risk; however, selection bias, counting errors, and lack of standardized assessment criteria have limited the clinical utility of angiogenesis quantitation. Therefore, we analyzed HCC angiogenesis with image cytometry using different methods and determined the correlation to prognosis. Tissue microarrays with 135 HCCs were CD31 and CD34 immunostained and quantitated with the Dako ACIS III Image Cytometer labeling index (LI) and Aperio Scanscope XT and MVD algorithm. LI and MVD were compared to each other and to pathologic features and prognosis (recurrence free survival). Using median cutoffs of microvessel quantitation, survival curve analysis showed a statistically significant difference between CD31 MVD algorithm measurement and prognosis (low MVD mean survival = 56.6 months and high MVD mean = 26.5 months; Log-Rank P = 0.0076). Survival was not significantly related to CD31 LI, CD34 LI or CD34 MVD. By linear regression, a direct correlation was observed between CD31 and CD34 using MVD (r = 0.45, P <0.0001), between CD31 MVD and CD31 LI (r = 0.55, P < 0.0001), and between CD31 LI and CD34 LI (r = 0.51, P < 0.0001). In addition, there was a weak but statistically significant relationship between CD31 MVD and CD34 LI (r = 0.25, P = 0.0050). Together, this data confirms previous studies linking angiogenesis to disease prognosis and suggests the utility of MVD image analysis algorithms.