摘要
目的 建立人子宫颈鳞状细胞癌细胞系HCC-0214,为子宫颈癌研究提供实验模型。方法无菌切除人子宫颈癌的手术标本,用组织块贴壁法体外培养,连续传代稳定生长后,绘制细胞生长曲线。采用光镜、电镜观察细胞形态,并进行细胞周期和染色体核形分析。用免疫组化法测定细胞系肿瘤标记物的(ER、PR、Keratine、PCNA)表达情况。结果组织块贴壁法体外培养获得人子宫颈鳞状细胞癌细胞系HCC-0214(简称H),细胞维持培养16个月,传代131代,生长稳定,群体倍增时间为35.48 h,细胞呈上皮镶嵌状贴壁生长,趋向复层生长,无接触抑制。超微结构显示,具有典型的桥粒结构和较多的张力原纤维。染色体数目35～156条,主流范围58～80条(64.8％),结构为人类染色体。细胞的肿瘤标记物(ER、PR、Keratine、PCNA)检测呈高表达,DNA指数为1.931。裸鼠移植瘤组织病理形态学与患者原始肿瘤一致,无血清培养成功。结论通过组织块贴壁法体外培养建立的人子宫颈鳞状细胞癌细胞系HCC-0214,与原发癌保持相同的生物学特性,体外连续传代16个月以上,细胞形态不变,生长周期恒定,可望作为一个稳定的细胞系。
Objective To establish a strain of human cervical carcinoma cell line and to provide a cervical carcinoma animal model. Methods The cervical carcinoma specimens incised aseptically were cultured in vitro by tissue culture methods, giving a tumor cell growth curve. Morphology of the cells was observed, with cell cycling analysis and chromosome analysis performed. The tumor markers (ER, PR, Keratine, PCNA) expressions of the cell line were detected by immuno-cytochemical technique. Results A human cervical carcinoma cell line HCC-0214(H) has been obtained by in vitro tissue culture methods. The cells have been maintained for 16 months through 131 passages, showing a stable growth with a population doubling time of 35.48 h and a tendency to pile up without contact inhibition. The ultrastructure showed typical desmosomes and numerous tonofilaments. Chromosome analysis revealed the number of chromosomes per cell varied from 35 - 156 with a stem-line number of 58 - 80 (64.8%). The morphology of chromosomes showed human tumor cell structure. The tumor markers (ER, PR, Keratine, PCNA) of the cells showed a high expression. The DNA index was 1.931 by flow cytometry (FCM). The histopathology of the transplanted tumors in nude mice was the same as the original tumor, though with none successful by serum culture. Conclusion A human cervical carcinoma cell line HCC-0214 established by tissue culture is identical to the primary cancer cell in biological characters. After the cells have been passaged for more than 16 months continually, their characteristics are still retained. Therefore, HCC-0214 may be used as a stable cell line.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2003年第1期43-46,共4页
Chinese Journal of Oncology
基金
国家自然科学基金重点资助项目(39830420)
