Kalra Neetu, Zhang Jingli, Thomas Anish, Xi Liqiang, Cheung Mitchell, Talarchek Jacqueline, Burkett Sandra, Tsokos Maria G, Chen Yuanbin, Raffeld Mark, Miettinen Markku, Pastan Ira, Testa Joseph R, Hassan Raffit
Thoracic and GI Oncology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
Laboratory of Pathology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
BMC Cancer. 2015 May 8;15:376. doi: 10.1186/s12885-015-1362-2.
The development and evaluation of new therapeutic approaches for malignant mesothelioma has been sparse due, in part, to lack of suitable tumor models.
We established primary mesothelioma cultures from pleural and ascitic fluids of five patients with advanced mesothelioma. Electron microscopy and immunohistochemistry (IHC) confirmed their mesothelial origin. Patient derived xenografts were generated by injecting the cells in nude or SCID mice, and malignant potential of the cells was analyzed by soft agar colony assay. Molecular profiles of the primary patient tumors, early passage cell cultures, and patient derived xenografts were assessed using mutational analysis, fluorescence in situ hybridization (FISH) analysis and IHC.
Primary cultures from all five tumors exhibited morphologic and IHC features consistent to those of mesothelioma cells. Mutations of BAP1 and CDKN2A were each detected in four tumors. BAP1 mutation was associated with the lack of expression of BAP1 protein. Three cell cultures, all of which were derived from BAP1 mutant primary tumors, exhibited anchorage independent growth and also formed tumors in mice, suggesting that BAP1 loss may enhance tumor growth in vivo. Both early passage cell cultures and mouse xenograft tumors harbored BAP1 mutations and CDKN2A deletions identical to those found in the corresponding primary patient tumors.
The mesothelioma patient derived tumor xenografts with mutational alterations that mimic those observed in patient tumors which we established can be used for preclinical development of novel drug regimens and for studying the functional aspects of BAP1 biology in mesothelioma.
恶性间皮瘤新治疗方法的开发和评估一直较少,部分原因是缺乏合适的肿瘤模型。
我们从5例晚期间皮瘤患者的胸水和腹水中建立了原发性间皮瘤培养物。电子显微镜和免疫组织化学(IHC)证实了它们的间皮来源。通过将细胞注射到裸鼠或SCID小鼠中产生患者来源的异种移植瘤,并通过软琼脂集落试验分析细胞的恶性潜能。使用突变分析、荧光原位杂交(FISH)分析和IHC评估原发性患者肿瘤、早期传代细胞培养物和患者来源的异种移植瘤的分子谱。
所有5个肿瘤的原代培养物均表现出与间皮瘤细胞一致的形态学和IHC特征。在4个肿瘤中均检测到BAP1和CDKN2A的突变。BAP1突变与BAP1蛋白表达缺失有关。3种细胞培养物均来源于BAP1突变的原发性肿瘤,表现出锚定非依赖性生长,并且在小鼠中也形成肿瘤,这表明BAP1缺失可能增强体内肿瘤生长。早期传代细胞培养物和小鼠异种移植瘤均具有与相应原发性患者肿瘤中相同的BAP1突变和CDKN2A缺失。
我们建立的具有与患者肿瘤中观察到的突变改变相似的间皮瘤患者来源的肿瘤异种移植瘤,可用于新药方案的临床前开发以及研究BAP1生物学在间皮瘤中的功能方面。