论文部分内容阅读
AIM:To investigate the progressive transformation ofimmortal cells of human fetal esophageal epitheliuminduced by human papUlomavirus,and to examine biologicalcriteria of sequential passage of cells,including cellularphenotype,proliferative rate,telomerase,chromosome andtumorigenicity.METHODS:The SHEE cell series consisted of immortalizedembryonic esophageal epithelium which was in malignanttransformation when cultivated over sixty passages withoutco-carcinogens.Cells of the 10th,31st,60th and 85thpassages were present in progressive development afterbeing transfected with HPV.Cells were cultivated in a cultureflask and 24-hole cultural plates.Progressive changes ofmorphology,cell growth,contact-inhibition,and anchorage-dependent growth characteristics were examined by phasecontrast microscopy.The cell proliferation rate was assayedby flow cytometry.The modal number of chromosomes wasanalyzed.HPV18E_6E_7 was detected by Western blot methodsand activities of telomerase were analyzed by TRAP.Tumorigenicity of cells was detected with soft agar platescultivated and with tumor formation in SCID mice.RESULTS:In morphological examination the 10th passagecells were in good differentiation,the 60th and 85th passagescells were in relatively poor differentiation,and the 31stpassage cells had two distinct differentiations.Thecharacteristics of the 85th and 60th passage cells wereweakened at contact-inhibition and anchorage-dependentgrowth.Karyotypes of four stages of cells belonged tohyperdiploid or hypotriploid,and bimodal distribution ofchromosomes appeared in the 31st and 60th passage cells.All of these characteristics combined with a increasing trend.The activities of telomerase were expressed in the latter threepassages.Four fourths of SCID mice in the 85th passagecells and one fourth of SCID mice in the 60th passage cellsdeveloped tumors,but the cells in the 10th and 31st passagedisplayed no tumor formation.CONCLUSION:In continual cultivation of fetal esophagea epithelial cells with transduction of HPV18E_6E_7,cells fromthe 10th to the 85th passage were changed gradually frompreimmortal,immortal,precancerous to malignantlytransformed stages,All of these changes were in a dynamicprogressive process.The establishment of a continuous lineof esophageal epithelium may provide a in vitro model ofcarcinogenesis induced by HPV.
AIM: To investigate the progressive transformation ofimmortal cells of human fetal esophageal epithelium induced by human papUlomavirus, and to examine biological criteria of sequential passage of cells, including cellular phenotype, proliferative rate, telomerase, chromosome and tumorigenicity. METHODS: The SHEE cell series consisted of immortalize depryonic esophageal epithelium which was in malignant transformation when cultivated over sixty passages withoutco-carcinogens. Cells of the 10th, 31st, 60th and 85th parass were present in progressive development after being transfected with HPV. Cells were cultivated in a cultureflask and 24-hole cultural plates. Progressive changes of morphology, cell growth, contact-inhibition, and anchorage-dependent growth characteristics were examined by phasecontrast microscopy. The cell proliferation rate was assayed by flow cytometry. The modal number of chromosomes was wasalyzed. HPV18E_6E_7 was detected by Western blot methods and activities of telomerase were analyzed by TRAP. Tumorigenicity of cells was detected with soft agar plates cultured and with tumor formation in SCID mice .RESULTS: In morphological examination the 10th passage cells were in good differentiation, the 60th and 85th passages cells were in poor poor differentiation, and the 31stpassage cells had two distinct differentiations. The characteristics of the 85th and 60th passage cells were weakened at contact-inhibition and anchorage-dependent growth. Karyotypes of four stages of cells belonged to hyperdiploid or hypotriploid, and bimodal distribution of chromosomes were found in the 31st and 60th passage cells. All of these characteristics combined with a increasing trend. The activities of telomerase were expressed in the latter three passages. Fourth fourth of SCID mice in the 85th passage cells and one fourth of SCID mice in the 60th passage cells depressed tumors, but the cells in the 10th and 31st passagedisplayed no tumor formation. CONCLUSION: In continual cultivation of fetal esophagea epithelial cells with transduction of HPV18E_6E_7, cells from the 10th to the 85th passage were changed gradually frompremortal, immortal, precancerous to malignantly transformed stages, All of these changes were in a dynamic progression process. The establishment of a continuous line of esophageal epithelium may provide in vitro models ofcarcinogenesis induced by HPV.