构建人类肾上腺皮质腺癌细胞系并用其验证过继免疫细胞治疗效果

Objective To establish a human adrenocortical carcinoma (ACC)cell line ACC-LWL, and investigate its cell phenotypes and expression of tumor associated antigens, to detect sensitivity of ACC-LWL against adoptive immune cells. Methods Fresh tumor tissues from the resection of a human ACC were primary cultured and passed generation to generation so as to achieve stable growth in vitro. For analyzing its biological characteristic, the cell cloning formation in soft agar, chromosome and tumorigenesis were tested. Flow cytometric analysis was carried out for phenotype analysis and RT-PCR examination for MN/CA9 and HLA-A2 genes expression. PBMC from 4 healthy donors (2 HLA-A2+ or 2 HLA-A3+) co-cultured with IL-2 (200 U/ml)and the tumor cell lysate of ACC-LWL (20 μg/ml) in vitro to generate CTL. After 14 day stimulation, CTLs were incubated with ACC-LWL with a E/T ratio of 10:1 for 4 hours at 37 ℃. Cytotoxicity was measured with MTr assay. Results One human ACC cell line had been established that showed the characteristics of malignant cells. The cells grew a solid tumor in nude mice. ACC-LWL had a high expression of MHC-Ⅰ, weakly expression of Her2/neu and MHC-Ⅱ, no expression of CD80 or CD86. RT-PCR showed that ACC-LWL expressed MN/CA9 and HLA-A2 genes. CTL, either HLA-A2+ or HLA-A3+, had the capability of killing ACC-LWL in vitro. Conclusion This newly established ACC would provide a useful target in vitro and in vivo for investigations related to human ACC.

Establishment and biological characteristics of a novel human adrenocortical carcinoma cell line and its sensitivity against CD8+ T cells