Induction of Human Breast Cancer Cell Apoptosis from G2/M Preceded by Stimulation into the Cell Cycle by Z-1,1-dichloro-2,3-diphenylcyclopropane
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We have shown previously that Z-1,1-dichloro-2,3-diphenylcyclopropane (a.k.a. Analog II, AII) inhibits human breast cancer cell proliferation regardless of estrogen receptor status or estrogen sensitivity, and that its cellular targets include microtubules. In the present study, we investigated the apoptosis-inducing effects of AII. MCF-7, MCF-7/LY2, and MDA-MB-231 cells all showed nuclear fragmentation in response to 100 μM AII when stained with Hoechst 33342 and examined by fluorescence microscopy. Pulsed field gel electrophoretic analysis showed that each of the cell lines also developed specific high molecular weight DNA fragments: a low level of 1–2 Mb fragments appeared after 6 hr, while 30–50 kb fragments accumulated subsequently. At 24 hr of drug exposure, the majority of cells became nonadherent, and the 30–50 kb fragments were restricted to detached MCF-7 and MDA-MB-231 cells. Both adherent and detached MCF-7/LY2 cells exhibited these fragments. A previous study by single-color (propidium) flow cytometry demonstrated that AII blocks MDA-MB-231 cells in G2/M of the cell cycle. More refined analyses in the present study showed this same result for MDA-MB-231 cells, but MCF-7 and MCF-7/LY2 cells did not reveal apparent drug-induced cell cycle block. AII demonstrated growth inhibitory, cell cycle-perturbing, and hypodiploidy-inducing activity against other human breast carcinoma lines, i.e. BT-20, CAMA-1, and SKBR-3, but no such actions in the non-tumorigenic, “normal” human breast epithelial line MCF-10A. Bromodeoxyuridine labeling and two-color flow cytometric analysis, however, suggested that AII caused stimulation into S phase, and that G2/M was the phase of the cell cycle from which cells apoptosed. AII caused cell rounding, detachment from the growth matrix, and nuclear shrinkage and fragmentation in parallel with biochemical changes. Cycloheximide inhibited AII-induced cell death, indicating that its toxicity requires de novo protein synthesis.
Medical Specialties | Medicine and Health Sciences | Osteopathic Medicine and Osteopathy
Balachandran, Raghavan; ter Haar, Ernst; Yalowich, Jack C.; Welsh, Manda J.; Grant, Stephen G.; and Day, Billy W., "Induction of Human Breast Cancer Cell Apoptosis from G2/M Preceded by Stimulation into the Cell Cycle by Z-1,1-dichloro-2,3-diphenylcyclopropane" (1999). College of Osteopathic Medicine Faculty Articles. 996.