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人乳头状瘤病毒18型E6E7基因诱导胎儿食管永生化上皮的生物学特性

Biological characteristics of human fetal esophageal epithelial cell line immortalized by the E6 and E7 gene of HPV type 18

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【作者】 沈忠英沈健蔡维佳岑山曾毅

【Author】 SHEN Zhongying, SHEN Jian, ZENG Yi, et al. Medical College of Shantou University Shantou 515031

【机构】 汕头大学医学院中国预防医学科学院肿瘤研究所

【摘要】 目的 SHEE细胞系是经人乳头状瘤病毒(HPV)18 型E6E7 基因诱导的永生化上皮细胞株,已传代超过50 代。研究胎儿食管上皮永生化的细胞株SHEE生物学特性,包括增殖,分化和调亡。方法 细胞于199 培养基培养,用光镜、电镜和荧光显微镜研究其生长率、形态和染色体分析;用流式细胞仪研究其细胞增殖动力学;用免疫组织化学方法研究Ki67 和角蛋白和用末端转移酶标记(TUNEL) 凋亡细胞。结果 细胞培养呈单层,锚定依赖和接触抑制生长。细胞生长曲线,增殖期3 ~8天,平顶期9 ~10 天,衰亡期10 天以后。增殖指数(PIx)34-0% ,分裂指数(MI)2-47% (1-20% ~4-80% ) ,凋亡指数(AI)1-30% ~6-90 % ,核染色体众数46 条为主(44 ~54 条) 和细胞周期DNA 呈2 倍体核型分布。电镜可见胞浆有张力原纤维,角蛋白免疫组织化学阳性证实鳞状上皮有分化特征。细胞凋亡可发生在增殖期,特别在衰亡期增多。结论 从生物学行为看来,SHEE细胞株接近于其来源细胞,胎儿食管上皮基底层,保存其增殖能力和分化的潜力。细胞死亡(包括细胞凋亡)是细胞群体扩增调节重要的因素,应作为研究生物学特性内容之一

【Abstract】 Objective Biological features including proliferation, differentiation and cell death of SHEE cell line, an immortalized epithelium of the fetal esophageal epithelium induced by HPV 18 E6E7 AAV, were studied. Methods SHEE cell line being cultured for more than 50 passages, were cultured in 199 growth medium and were examined by light, electron and fluorescence microscopy for growth rate, morphological features and chromosome analysis, by flow cytometry for cell proliferative dynamics, by immunohistochemistry for Ki67 and cytokeratin, and by terminal DNA transferase label (TUNEL) for apoptosis. Results At the 20th passage, the SHEE cell remained monolayer, anchorage dependent and attachment inhibited growth. The growth curve showed proliferative phase 3~8th days, top plate phase 9~10th days and attenuative phase after 10th days. Proliferative index(PIx)34.0%, mitotic index (MI)2.74%(1.20%~4.80%), apoptotic index (AI)1.30%~6.90%, chromosome analysis mainly 46 (44~54/nucleus) and DNA distribution in diploidy were calculated and described. The tonofilament expression in cell cytoplasm by electron microscopy and positive reaction of cytokeratin by immunochemistry showed differentiative character of squamous epithelium. The cell apoptosis occured in the proliferative phase and especially increased in attenuative phase.Conclusion Of biological behaviors, the SHEE cells are close to the basal cells of their original fetal esophageal mucosa keeping proliferative and differentiative potency. This study suggests that the cell death (including cell apoptosis) may be an important factor in studying of cell growth regulation and it may be an research area for cellular biological behaviors.

【基金】 国家自然科学基金
  • 【文献出处】 中华实验和临床病毒学杂志 ,CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY , 编辑部邮箱 ,1999年03期
  • 【分类号】R373
  • 【被引频次】14
  • 【下载频次】104
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