At the 20th passage, SHEE cells overcame the senescence and apoptosis and restored their proliferative activity with expression of telomerase and hTERT at low levels, but the telomere length shortened continuously to the lowest of 3 kb. From the 12th to the 16th passages, many senescent and apoptotic cells appeared, and the telomere length sharply shortened from 23 kb to 17 kb without expression of hTERT and telomerase activity. RESULTS: SHEE cells from the 6th to 10th passages showed cellular proliferation with a good differentiation. The cell apoptosis was examined by electron microscopy (EM) and TUNEL label. DNA content in cell cycle was detected by flow cytometry. Expressions of subunits of telomerase, hTR and hTERT, were assessed by RT-PCR. ![]() ![]() Morphological phenotype of cultured SHEE cells from the 6th to 30th passages, was examined by phase contrast microscopy, the telomere length was assayed by Southern blot method, and the activity of telomerase was analyzed by telomeric repeat amplification protocol (TRAP). METHODS: The transgenic cell line of human fetal esophageal epithelium (SHEE) was established with E 6E 7 genes of human papillomavirus (HPV) type 18 in our laboratory. AIM: To search for the biomarker of cellular immortalization, the telomere length, telomerase activity and its subunits in cultured epithelial cells of human fetal esophagus in the process of immortalization.
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