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Author Ji Young Kwon, M.D., Yoon Sung Jo, M.D., Ye Hoon Choi, M.D., Jong Gyu Chang, M.D., Ki Sung Ryu, M.D., Jong Gu Rha, M.D. and Ku Taek Han, M.D.
Place of duty Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea, Seoul, Korea
Title Detection of p16INK4A in the Mixed Cell Populations of Normal Peripheral Blood Mononuclear Cells and Cervical Cancer Cell Lines
Publicationinfo Cancer Research and Treatment 2003 Jun; 035(03): 254-260.
Key_word p16INK4A, Cytokeratin, Cervical cancer cell lines, Flow cytometry
Full-Text
Abstract Purpose: Human papilloma viruses (HPVs) play a central role in the pathogenesis of neoplastic lesions of the uterine cervix. The viral oncoprotein HPV E6 degrades the p53 protein, and the HPV E7 protein inactivates pRB and increases the expression of the CDK inhibitor, p16INK4A. We investigated the usefulness of p16INK4A as a biologic marker for the cervical dysplastic and neoplastic cells. Materials and Methods: We examined the expression of p16INK4A and cytokeratin in a mixed population of normal peripheral blood mononuclear cells (PBMC) and the cervical cancer cell lines (HeLa, SiHa, and CasKi) using flow cytometry. Results: The DNA indices of the HeLa, SiHa and CasKi cell lines were 1.89, 1.53 and 1.75, respectively, indicating that these cells are aneuploid cells. Furthermore, the positive rate of p16INK4A expression was 86.7% for the HeLa mixed population, 85.6% for the SiHa mixed population, and 92.2% for the CasKi mixed population. According to the FL3A vs FL3W histogram, electrical gating of the HeLa, SiHa and CasKi mixed populations showed the expression levels of both cytokeratin and p16INK4A to be identical, at 86.6%, 84.8% and 85.0%, respectively. These findings revealed that almost all cells selected through electrical gating were cervical cancer cells originating from the epithelium and which expressed cytokeratin and p16INK4A. On the other hand, when each mixed population was electrically gated for normal PBMC, we found that the PBMCs expressed neither cytokeratin nor p16INK4A. Conclusion: Using flow cytometry, we observed the enhanced expression of p16INK4A in cervical cancer cell lines. These results suggest the usefulness of p16INK4A for the selective detection of cervical dysplastic and cancer cells in the liquid-based samples, which are taken from the cervices and contaminated with blood and stromal cells. (Cancer Research and Treatment 2003;35:254 260)