Development of a novel method for measuring in vivo breast epithelial cell proliferation in humans

Abstract

Cell proliferation plays an important role in all stages of carcinogenesis. Currently, no safe, direct, in vivo method of measuring breast epithelial cell (BEC) proliferation rates in humans exists. Static immunohistochemical markers of cell proliferation, such as Ki-67 and PCNA indices, have technical limitations including high inter-lab variability, inaccuracy in the presence of agents that cause G1/S cell cycle block and inadequate sensitivity in post-menopausal women with low BEC proliferation rates. We describe here a safe, direct method of measuring BEC proliferation rates in vivo in women using heavy water ((2)H(2)O) labeling coupled with mass spectrometric analysis. Proliferation of normal and tumor BEC was measured from breast tissue biopsies in women undergoing mastectomy (n = 11) and normal BEC from healthy volunteers (n = 16). Women took heavy water (50-150 ml per day) for 1-4 weeks. Pre-menopausal women had significantly higher proliferation rates than post-menopausal women (0.7 +/- 0.1 versus 0.2 +/- 0.1 new cells per day, respectively), and tumor BEC had different proliferation rates than normal BEC from the same breast. The method is analytically reproducible and remains sensitive in the range of low proliferation rates. In summary, this novel method of measuring BEC proliferation in vivo holds promise for assessing the effects of anti-proliferative chemopreventive and chemotherapeutic agents.