Roberts Darren L, O'Dwyer Sarah T, Stern Peter L, Renehan Andrew G
Immunology Group, Paterson Institute for Cancer Research, The University of Manchester, Manchester, M20 4BX, UK.
Institute of Cancer Sciences, The University of Manchester, Manchester Academic Health Science Centre, The Christie NHS Foundation Trust, Manchester M20 4BX, UK.
Oncotarget. 2015 May 10;6(13):10786-800. doi: 10.18632/oncotarget.3198.
Pseudomyxoma peritonei (PMP) is a rare tumor of appendiceal origin. Treatment is major cytoreductive surgery but morbidity is high. PMP is considered chemo-resistant; its molecular biology is understudied; and presently, there is no platform for pre-clinical drug testing. Here, we performed exon array analysis from laser micro-dissected PMP tissue and normal colonic epithelia. The array analysis identified 27 up-regulated and 34 down-regulated genes: candidate up-regulated genes included SLC16A4, DSC3, Aldolase B, EPHX4, and ARHGAP24; candidate down-regulated genes were MS4A12, TMIGD1 and Caspase-5. We confirmed differential expression of the candidate genes and their protein products using in-situ hybridization and immuno-histochemistry. In parallel, we established two primary PMP cell lines, N14A and N15A, and immortalized with an SV40 T-antigen lentiviral vector. We cross-checked for expression of the candidate genes (from the array analyses) using qPCR in the cell lines and demonstrated that the gene profiles were distinct from those of colorectal tumor libraries and commonly used colon cell lines. N14A and N15A were responsiveness to mitomycin and oxaliplatin. This study characterizes global gene expression in PMP, and the parallel development of the first immortalized PMP cell lines; fit for pre-clinical testing and PMP oncogene discovery.
腹膜假黏液瘤（PMP）是一种罕见的阑尾源性肿瘤。治疗方法主要是细胞减灭术，但发病率很高。PMP被认为对化疗耐药；其分子生物学研究较少；目前，尚无临床前药物测试平台。在此，我们对经激光显微切割的PMP组织和正常结肠上皮进行了外显子阵列分析。阵列分析鉴定出27个上调基因和34个下调基因：上调候选基因包括SLC16A4、DSC3、醛缩酶B、EPHX4和ARHGAP24；下调候选基因有MS4A12、TMIGD1和半胱天冬酶-5。我们通过原位杂交和免疫组织化学证实了候选基因及其蛋白产物的差异表达。同时，我们建立了两个原发性PMP细胞系N14A和N15A，并用SV40 T抗原慢病毒载体使其永生化。我们在细胞系中使用qPCR对（来自阵列分析的）候选基因的表达进行了交叉核对，结果表明这些基因谱与结直肠肿瘤文库和常用结肠细胞系的基因谱不同。N14A和N15A对丝裂霉素和奥沙利铂有反应。本研究对PMP中的整体基因表达进行了表征，并首次平行开发了永生化PMP细胞系；适用于临床前测试和PMP癌基因的发现。
