为筛选药物建立用作临床前工具的原发性子宫内膜癌细胞培养物模型：方法与特征

　　ESTABLISHMENT OF PRIMARY ENDOMETRIAL CARCINOMA CELL CULTURES AS A PRECLINICAL TOOL FOR DRUG SCREENING: METHODS AND CHARACTERIZATION

　　S. Schrauwen¹, L. Coenegrachts¹, D. Garcia¹, C. Luyten¹, G. Verbist¹, I. Vergote¹, D. Lambrechts², F. Amant¹

　　¹Oncology, ²Versalius Research Center, KU Leuven, Leuven, Belgium

　　Background: Treatment options for primary advanced and recurrent endometrial cancer (EC) are limited, making development of new treatment strategies essential.

　　Methods: After informed consent, fresh endometrial tumor biopsies were collected. Primary cell cultures were established after collagenase digestion and fibroblast depletion. They were validated by (i) measurement of telomerase activity, (ii) dynamic monitoring of cell proliferation (xCELLigence), and (iii) immunocytochemical stainings. In vivo tumorigenicity was determined in a s.c. xenograft model. Mutation analysis (Sequenom) was performed to detect hotspot mutations in genetic pathways that are known to be affected in EC (e.g. PI3K/PTEN/AKT and EGFR/KRAS). The mutation profile of the primary tumor, cell culture and xenograft was compared.

　　Results: Success rate for grade 2 and 3 tumors was ~30%. Telomerase activity of all cell cultures was similar to this of the HEC-1A cell line. Doubling time ranged between 19-28h. All cultures were negative for CD90 and a-SMA, positive for cytokeratin, whereas vimentin expression varied among cultures. In vivo tumorigenicity analysis showed tumor take rates between 60-100%. Mutational profiling identified PTEN, PIK3CA, KRAS, NRAS, BRAF and p53 mutations, which remained identical in the primary tumor, the cell culture, and xenograft.

　　Conclusion: We here present the first evidence for the establishment of primary EC cell cultures, providing an alternative, primary tumor-based model for in vitro and in vivo (targeted) therapeutic drug screening in EC.

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