1. Growth-inhibitory Effects Of Difluoromethylornithine (DFMO), Genistein In Immortalized Cervical Epithelial Cells, Cancer Cells And Xenografts In Nude Mice, And Their Regulation Mechanisms In Cells Cycle 2. Detection Of Physical Status Of Human Papillom

Growth-inhibitory Effects of Difluoromethylornithine (DFMO), Genistein in Immortalized Cervical Epithelial Cells, Cancer Cells and Xenografts in Nude Mice, and Their Regulation Mechanisms in Cell CycleObjectiveTo observe the effects of difluoromethylornithine (DFMO), genistein on the growth inhibition in human immortalized cervical epithelial cell line, cervical cancer cell lines, and xenografts in nude mice. And to investigate their regulation mechanisms in cell cycle. Methods(1) Growth inhibition analysis: One human cervical epithelial cell line (H8), and two cervical carcinoma cell lines (CaSki, HeLa) were studied. The effects of DFMO or Genistein on growth inhibition were tested in monolayer culture, the growth curves and concentrations required for a 50% growth inhibition (IC₅₀) with a 5-day treatment were determined by the sulforhodamine B (SRB) assay.(2) Cell cycle analysis. Cellular DNA content was measured and analyzed using flow cytometry of Coulter EPICS XL.(3) Immunofluorescence analysis. Primary antibodies were monoclonal antibodies of p21, cyclin D1, CDK4. FITC-labeled secondary antibodies were used. The fluorescence was measured by Coulter EPICS XL.(4) Reverse Transcription (RT) PCR. The mRNA expressions of cyclin D1, cyclin B1, CDK4, cdc2 and p21 were determined by RT-PCR semi-quantitatively.(5) Malignant transformation. TPA transformed the immortalized cervical epithelial cells (H8), for 40 days.(6) Prevention of Malignant transformation. DFMO or Genistein was added with...