A comparative study of telomerase activity and malignant phenotype in multistage carcinogenesis of esophageal epithelial cells induced by human papillomavirus.
A comparative study of telomerase activity and malignant phenotype in multistage carcinogenesis of esophageal epithelial cells induced by human papillomavirus.
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人乳头瘤病毒诱导食管上皮细胞多期癌变中端粒酶活性与恶性表型的比较研究
DOI：
10.3892/ijmm.8.6.633
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发表时间：
2001-12
期刊：
影响因子：
--
通讯作者：
Zeng Y
中科院分区：
文献类型：
--
作者：
Shen ZY;Xu LY;Li C;Cai WJ;Shen J;Chen JY;Zeng Y
为了研究多步骤癌变的某些特征，我们研究了HPV诱导的食管上皮细胞永生化、癌前和恶性阶段的端粒酶活性和恶性表型。用人乳头瘤病毒（HPV）18型E6 E7基因诱导建立永生化人胎食管上皮细胞系（SHEE）。从SHEE 61的非贴壁生长集落中选择并扩增的第10代（SHEE 10）、第31代（SHEE 31）、第61代（SHEE 61）和SHEE 61 A中的细胞进行如下检查：通过电子显微镜观察细胞形态;流式细胞术检测细胞周期，TRAP法检测端粒酶活性，致瘤性检测包括软琼脂贴壁非依赖性生长、SCID小鼠和裸鼠体内成瘤实验以及Western blot检测HPV 18 E6 E7癌蛋白。SHEE 10细胞的形态表现出良好的分化，SHEE 60和SHEE 61 A细胞的分化相对较差，SHEE 31细胞以两种不同的方式分化。SHEE 31、SHEE 61和SHEE 61 A细胞端粒酶活性明显增强，而SHEE 10细胞端粒酶活性不明显。SHEE 61和SHEE 61 A细胞的接触抑制减弱，贴壁非依赖性生长增加。SHEE 61细胞接种SCID小鼠和裸鼠后，前两代细胞均未成瘤; SHEE 61细胞在4只SCID小鼠中均成瘤，而在裸鼠中均未成瘤; SHEE 61 A细胞在两种免疫缺陷小鼠中均成瘤。Western blotting检测HPV 18 E6 E7 DNA阳性。在HPV的致癌过程中，SHEE 31细胞处于具有端粒酶活性的永生化状态。事实上，SHEE 61细胞保持永生化，也表现出锚定非依赖性生长，揭示了癌前特征; SHEE 61 A细胞表现出恶性转化，在小鼠中形成肿瘤。结果表明，端粒酶活性、非贴壁生长和裸鼠成瘤分别是肿瘤永生化、癌前病变和恶性的指标。
To examine certain characteristics of multistep carcinogenesis, we studied telomerase activity and malignant phenotypes in the immortal, premalignant and malignant stages of esophageal epithelial cells induced by HPV. An immortalized human fetal esophageal epithelial cell line (SHEE) was induced by E6E7 genes of human papillomavirus (HPV) type 18. Cells in the 10th passage, (SHEE10), 31st passage (SHEE31), 61st passage (SHEE61) and SHEE61A which were selected and expanded from anchorage-independent growth colonies of SHEE61, were examined as follows: cell morphology by electron-microscopy; the cell cycle by flow cytometry, telomerase activity by TRAP assay, tumorigenic detection including anchorage-independent growth by soft agar culture and tumor formation by inoculating cells into SCID and nude mice, and detection of HPV18 E6E7 oncoprotein by Western blot. The morphology of the SHEE10 cells exhibited good differentiation, the SHEE60 and SHEE61A cells were relatively poorly differentiated, and the SHEE31 cells were differentiated in two distinct ways. The telomerase was activated in SHEE31, SHEE61 and SHEE61A, but not in SHEE10 cells. SHEE61 and SHEE61A cells were weakened in contact-inhibition and increased in anchorage-independent growth. Inoculated into SCID and nude mice, the cells of the earlier two passages could not develop tumors; the SHEE61 developed one tumor in four SCID mice, but not in nude mice, and the SHEE61A cells developed tumors in both strains of immunodeficient mice. HPV18 E6E7 DNA detection by Western blotting was positive in all cell passages. In the process of carcinogenesis by HPV, the cells of SHEE31 are in an immortalized state with telomerase activity. The fact that SHEE61 cells remained immortalized and also demonstrated anchorage-independent growth, reveals premalignant character; the cells of SHEE61A exhibited malignant transformation with tumor formation in mice. The results revealed that the telomerase activity, anchorage-independent growth and tumor formation in nude mice are the indicators for immortalization, premalignancy and malignancy, respectively.