The Study Of Human Ovarian Cancer Cell Line Treated With Anti-Tumor Agents By Laser Scanning Cytometry.
A. Yoshizaki*1, T. Honda1, A. Kurose2, K. Sasaki2 and I. Nishiya1 Iwate Medical University, Dept of Ob & Gyn1, Dept of 1st Pathology2, Japan.
[Aim] Here we will present the cell cycle changes and the morphological changes which are produced by paclitaxel and CDDP(P) in human ovarian cancer cell line using laser scanning cytometry (LSC).
[Materials and Methods] P resistant human ovarian cancer cell line (KFr13) which was kindly provided by Dr. Kikuchi, were exposed to 10-5, 10-6, 10+7M of Paclitaxel(T) and 1 µg/mL of P at day 3 of cell culture for 3, 6, 12 and 24 hours. DNA histograms and morphological features of cells stained by 50 µg/mL PI containing 100 µg/mL RNase were analysed and observed by LSC. [Results] T began to produce G2+M accumulation by 3hours. G2+M accumulation was significant at 12hours. This G2+M peak was containing G2 phase cells, M (including metaphase) phase cells and apoptotic cells. T induced apoptosis at G2 phase. The % of apoptotic cells were 9-12% at 3hours. 17-20% at 6hours and 56-65% at 24hours comparing less than 5% in control. P showed late S and G2+M accumulations in DNA histogram. These cell accumulations were consisted by the cells which were chromatin riched and had enlarged nuclei. Few M phase cells and no apoptotic cells were observed. [Summary] LSC was able to offer each cell features consisting the histograms and cytograms. This will enable us to analyse cells during anti-tumor agent treatment in vitro and in vivo more precisely.