人喉癌细胞系的建立及生物学特性研究

Establishment and Charateristics Research of Human Laryngeal Squamous Carcinoma Cell Line

目的建立2种人喉癌细胞系，确定其生物学特性。方法取喉癌患者原发病灶和淋巴结转移灶组织，用组织块培养法进行原代培养。20代后观察细胞形态，绘制生长曲线图，用免疫组织化学染色方法鉴定细胞分子特征，测定细胞周期和软琼脂集落形成率。结果建立了2个人喉癌细胞系，分别命名为CCC-HLS和CCC-HLSLN，已传至第25代。CCC-HLS细胞形态以梭形为主；CCC-HLSLN细胞呈多形性，少数细胞有细长的突起，末端有分叉。2种细胞角蛋白（CK）免疫组织化学染色均为阳性。2种细胞传代后，2～4d为对数生长期，第7天开始死亡，CCC-HLS细胞倍增时间为33．9h，CCC-HLSLN细胞倍增时间为30h。细胞周期分析表明CCC-HLS细胞Go／G1期为54．7％，G2／M期为5．2％，S期为40．1％。CCC-HLSLN细胞G0／G1期为50．6％，G2／M期为27．4％，S期为22．0％。CCC-HLS细胞软琼脂集落形成率为1．23％，CCC-HLSLN软琼脂集落形成率为2．58％。结论建立的2种人喉癌细胞系可用于喉癌的基础研究。

Objective To establish human laryngeal squamous carcinoma cell line and define its biological eharateristies. Methods Primary tumor tissue and the metastatic lymph node of laryngeal squamous carcinoma from a patient were used for culture with the regular small mass method. Alter 20 passages, their morphologies, growth curves, immunohistochemistry staining, cell cycles and colony- forming efficiency in soft agar were determined. Results Two human laryngeal squamous carcinoma cell lines were designated CCC-HLS and CCC-HLSLN respectively. CCC-HLS cells were main spindle, while CCC-HLSLN cells demonstrated various morphologies and some often had thin and long projection with branches in the end. Both of the cells were CK-positive. From the second day to the forth day, it was in the exponent growing stage for both cell lines and they began to die on the seventh day. The population doubling time of CCC-HLS cells was 33. 9 h and that of CCC-HLSLN cells was 30 h. 54.7% CCC-HLS cells were in Go/G1 , 5. 2% in G2/M and 40. 1% in S, and 50. 6% CCC-HLSLN cells in Go/G1, 27. 4% in G2/M and 22.0% in S. The colony-forming rate of CCC-HLS cells was 1. 23% and that of CCC-HLSLN cells was 2. 58%. Conclusion We established two human laryngeal squamous carcinoma cell lines and studied their biological characteristics systematically. These cells can be used for basic research on laryngeal carcinoma.