Increased Midkine Expression in Hepatocellular Carcinoma Immunohistochemical and In Situ Hybridization Analyses

c Context.—Midkine (MK) is a novel heparin-binding growth factor whose gene was identified in embryonal carcinoma cells in early stages of retinoic acid‐induced differentiation. Objective.—To examine the overexpression of MK in hepatocellular carcinoma (HCC). Methods.—Seventy-seven primary HCC specimens from patients aged 17 to 72 years (63 men and 14 women) were examined. Histologically, 16 cases of HCC were classified as the well-differentiated type, 50 cases as the moderately differentiated type, and 11 cases as the poorly differentiated type. Immunohistochemical analysis was performed using a rat immunoglobulin G2a monoclonal antibody against the carboxyl terminal region of human MK. In situ hybridization was also performed on 20 HCC samples. Results.—We successfully applied this monoclonal antibody against MK to analyze archival paraffin sections. The cancer tissues showed a positive reaction to this antibody, in which there was an intense reaction in their cytoplasm. Approximately one third of the individuals with HCC (26/ 77) had tumor cells that expressed MK, and these were classified into the following types: moderately differentiated (20/50), well differentiated (3/16), and poorly differentiated (3/11). The in situ hybridization analysis revealed that the signals of MK transcripts were found in the cytoplasm of the cancer cells; the distribution and localization of the MK transcripts’ signals determined by in situ hybridization analysis were similar to those obtained by immunohistochemical analysis. Conclusions.—Hepatocellular carcinoma expressed increased MK at the messenger RNA and protein level. (Arch Pathol Lab Med. 2000;124:848‐852)