具有不同转移潜能的前列腺癌细胞亚系的分离鉴定

目的 克隆具有不同转移潜能的癌细胞亚系,并应用体内外实验对各亚系的侵袭转移能力进行检测。方法 应用倍比稀释法对前列腺癌细胞系ＰＣ－３Ｍ进行单细胞克隆,并应用体内外实验包括生长曲线,人工基底膜侵袭实验,软琼脂集落形成,裸鼠异种接种及自发性转移实验检测各亚系的生物学特性及侵袭转移能力。结果 从前列腺癌细胞系ＰＣ－３Ｍ成功分离建立了４个具有不同转移潜能的亚系,经体内外多项实验检测证明：１Ｅ８为高转移亚系

Isolation and characterization of human prostate cancer cell subclones with different metastatic potential

OBJECTIVE: To isolate and characterize cancer cell subclones with different metastatic potential from human metastatic prostate cancer cell line (PC-3M). METHODS: Using limited dilution, in vitro growth, Matrigel invasion assay, soft agar cloning and in vivo tumorigenicity and spontaneous metastasis assay in nude mice, were isolated and characterized four subclones with different metastatic potential. RESULTS: Four subclones derived from PC-3M were 1E8, 2E7, 2B6, 2B4. Each subclone exhibited a different metastatic potential when inoculated into nude mice. Among these subclones, 1E8 expressed as the highly metastatic phenotype, with 100% metastasis frequency 5 weeks after subcutaneous inoculation into nude mice, whereas 2B4 was not metastatic. In vitro 1E8 was found to be the most highly invasive cell line in Matrigel invasive assay (98 +/- 24) and had the most clones in soft agar cloning assay (265 +/- 39) while 2B4 was found to have the least invasive abilities (12 +/- 4) and the least clones (137 +/- 14). CONCLUSION: Successful establishment of these subclones with different metastatic potential may be valuable for further study on the molecular mechanisms of cancer metastasis and cloning of cancer metastasis-related genes.