Comparative flow cytometric analysis of proliferating cell nuclear antigen (PCNA) antibodies in human solid neoplasms

Proliferating cell nuclear antigen (PCNA), an auxiliary protein of DNA polymerase delta, associated with DNA synthesis, is increasingly used to determine tumor growth rate. To determine the growth fraction by PCNA, various antibodies and fixation procedures are being used. We assessed the flow-cytometric measurement of PCNA using three monoclonal antibodies (PC10, 19F4, and 19A2) and two fixation protocols (paraformaldehyde and methanol) in tissue-culture cell lines and human solid neoplasms to determine their potential clinical application. Thirty-one solid tumors and four normal tissues were analyzed, along with MOLT-4 and HL-60 cell lines as positive controls and normal peripheral blood lymphocytes as a negative control. PC10 with methanol fixation consistently detected higher PCNA positivity in human solid neoplasms than 19F4 and 19A2. Using PC10, the differences in positivity among fixation methods occurred in the G0/1 phase, not in S + G2M, of the cell cycle. No correlation was found between PCNA positivity and tumor grade and DNA ploidy in tumors analyzed. A statistical correlation was found between overall PCNA positivity and RNA content as determined by acridine orange analysis. The growth fraction by PCNA in solid neoplasms was most reliably determined by PC10 with methanol fixation.