Baker Amanda F, Malm Scott W, Pandey Ritu, Laughren Cindy, Cui Haiyan, Roe Denise, Chambers Setsuko K
University of Arizona Cancer Center and College of Medicine, Tucson, Arizona,
Cancer Microenviron. 2015 Apr;8(1):45-56. doi: 10.1007/s12307-015-0166-x. Epub 2015 May 22.
A panel of nine hypoxia regulated genes, selected from a previously published fifty gene panel, was investigated for its ability to predict hypoxic ovarian cancer phenotypes. All nine genes including vascular endothelial growth factor A, glucose transporter 1, phosphoglycerate mutase 1, lactate dehydrogenase A, prolyl 4-hydroxylase, alpha-polypeptide 1, adrenomedullin, N-myc downstream regulated 1, aldolase A, and carbonic anhydrase 9 were upregulated in the HEY and OVCAR-3 human ovarian cell lines cultured in vitro under hypoxic compared to normoxic conditions as measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). The gene panel was also elevated in HEY xenograft tumor tissue compared to HEY cells cultured in normoxia. The HEY xenograft tissue demonstrated heterogeneous positive immunohistochemical staining for the exogenous hypoxia biomarker pimonidazole, and the hypoxia regulated protein carbonic anhydrase IX. A quantitative nuclease protection assay (qNPA) was developed which included the nine hypoxia regulated genes. The qNPA assay provided similar results to those obtained using qRT-PCR for cultured cell lines. The qNPA assay was also evaluated using paraffin embedded fixed tissues including a set of five patient matched primary and metastatic serous cancers and four normal ovaries. In this small sample set the average gene expression was higher in primary and metastatic cancer tissue compared to normal ovaries for the majority of genes investigated. This study supports further evaluation by qNPA of this gene panel as an alternative or complimentary method to existing protein biomarkers to identify ovarian cancers with a hypoxic phenotype.
从先前发表的50个基因组合中挑选出一组9个缺氧调节基因，对其预测缺氧型卵巢癌表型的能力进行了研究。通过定量逆转录聚合酶链反应（qRT-PCR）检测发现，与常氧条件相比，在体外低氧培养的HEY和OVCAR-3人卵巢癌细胞系中，包括血管内皮生长因子A、葡萄糖转运蛋白1、磷酸甘油酸变位酶1、乳酸脱氢酶A、脯氨酰4-羟化酶α多肽1、肾上腺髓质素、N- myc下游调节因子1、醛缩酶A和碳酸酐酶9在内的所有9个基因均上调。与在常氧条件下培养的HEY细胞相比，HEY异种移植肿瘤组织中的该基因组合也有所升高。HEY异种移植组织对外源性缺氧生物标志物匹莫硝唑和缺氧调节蛋白碳酸酐酶IX表现出异质性阳性免疫组化染色。开发了一种包括9个缺氧调节基因的定量核酸酶保护分析（qNPA）方法。qNPA分析对培养细胞系获得的结果与qRT-PCR相似。还使用石蜡包埋的固定组织对qNPA分析进行了评估，这些组织包括一组5例患者匹配的原发性和转移性浆液性癌以及4个正常卵巢。在这个小样本集中，对于大多数研究的基因，原发性和转移性癌组织中的平均基因表达高于正常卵巢。本研究支持通过qNPA对该基因组合进行进一步评估，作为现有蛋白质生物标志物的替代或补充方法，以识别具有缺氧表型的卵巢癌。
