舌癌耐药细胞株的建立及癌干细胞特性的实验观察

Establishment of a drug-resistant tongue cancer cell line and analysis of its cancer stem cell characteristics

目的建立舌癌耐药细胞株,并观察耐药细胞株癌干细胞的相关生物学特性。方法以低浓度阿霉素持续增量诱导法建立TCA-8113舌癌耐药细胞株,并对亲代与舌癌耐药细胞株进行癌细胞基本特性、癌干细胞相关细胞学特性的检测,分别以计数法检测癌细胞生长曲线、倍增时间以及用MTT法测定耐药倍数,并进行单个细胞成克隆实验、平板克隆形成实验和悬浮球形成实验。结果 TCA-8113舌癌耐药细胞为"铺路石"状上皮样生长方式,但其中异形细胞略多,耐药倍数为22倍;生长曲线提示其增殖速度明显低于亲本TCA-8113舌癌细胞,且倍增时间增加,但在细胞周期中各期细胞比例未见明显改变;TCA-8113舌癌耐药细胞克隆形成率、单个细胞培养成株率均明显低于其亲本TCA-8113舌癌细胞(P﹤0.05)。结论低浓度阿霉素持续增量法可诱导形成舌癌耐药细胞株,但舌癌耐药细胞株的癌干细胞特征并不明显。

Objective To establish a drug-resistant tongue cancer cell line and analyze its cancer stern cell （CSC）characteristics. Methods Tongue cancer cell line TCA-8113 was induced by adriamycin （ADM）and used to establish the drug-resistant cell line TCA-8113/ADM.Basic characteristics of both the parental cell line TCA-8113 and the drug-resistant cell line TCA-8113/ADM were analyzed and compared, including cell growth, doubling time, cell cycle, resistance index（ RI ）, single-cell culture, colony formation and formation of sphere cells. Results The growth pattern of TCA-8113/ADM cells was cobblestone-like, with cultures showing sig- nificantly more atypical cells than those of parental cells.TCA-8113/ADM cells had an RI of 22, and their rates of cell proliferation, cell doubling, colony formation and sphere cell formation were slower than those of the parental cells. Conclusion Sustained incremental exposure to low concentrations of ADM can induce a drug-resistant tongue cancer cell line,but its CSC characteristics are not obvious.