Li Jie, Mitani Yohitsugu, Rao Pulivarthi H, Perlaky Laszlo, Liu Bin, Weber Randal S, El-Naggar Adel K
The University of Texas, M.D. Anderson Cancer Center, Department of Pathology, United States.
Baylor College of Medicine, Department of Pediatrics, Texas Children's Cancer and Hematology Centers, United States.
Oral Oncol. 2017 Jun;69:108-114. doi: 10.1016/j.oraloncology.2017.04.007. Epub 2017 May 2.
To develop and characterize in vitro salivary duct carcinoma as a surrogate for functional studies.
Cells were dispersed from tumor tissue fragments under sterile conditions in RPMI media. Disassociated cells were cultivated, immortalized with hTERT and propagated for more than 100 passages. Morphologic, linage, cytogenetic and genomic analyses were performed on different passages of cell line and primary tumor. Soft agar growth was performed.
Analysis of cytomorphologic features, growth characteristics and lineage specific markers expression confirmed the epithelial derivation and the neoplastic nature of the cell line. DNA STRs analysis showed identical match of both cell line and primary tumor. Cultivated cells expressed Androgen Receptor (AR), PTEN, and EFGR proteins and the AR-V7 isoform transcript. Comparative exome-sequencing identified common mutated genes in both cell line and primary tumor. In-vitro colony formation of late passages is established.
We report the development of the first human salivary duct carcinoma cell line (MDA-SDC-04) that retains critical biological and genomic features of the donor tumor.
建立体外唾液腺导管癌模型并对其进行表征,以用于功能研究。
在无菌条件下,将肿瘤组织碎片中的细胞分散于RPMI培养基中。分离出的细胞进行培养,用hTERT使其永生化,并传代培养超过100代。对细胞系和原发性肿瘤的不同传代进行形态学、谱系、细胞遗传学和基因组分析。进行软琼脂生长实验。
细胞形态学特征、生长特性和谱系特异性标志物表达分析证实了该细胞系的上皮来源和肿瘤性质。DNA STRs分析显示细胞系和原发性肿瘤完全匹配。培养的细胞表达雄激素受体(AR)、PTEN和EFGR蛋白以及AR-V7异构体转录本。比较外显子组测序确定了细胞系和原发性肿瘤中的共同突变基因。晚期传代细胞在体外形成集落。
我们报告了首个保留供体肿瘤关键生物学和基因组特征的人唾液腺导管癌细胞系(MDA-SDC-04)的建立。