Division of Pathology, Exploratory Research & Clinical Trial Center, National Cancer Center, Kashiwa, Japan.
Department of Hepatobiliary and Pancreatic Surgery, National Cancer Center Hospital East, Kashiwa, Japan.
BMC Gastroenterol. 2020 Dec 9;20(1):411. doi: 10.1186/s12876-020-01556-w.
Stromal fibroblasts associated with pancreatic ductal adenocarcinoma (PDAC) play an important role in tumor progression through interactions with cancer cells. Our proposed combination strategies of in vitro and in silico biomarker screening through a cancer-stromal interaction model were previously identified several actin-binding proteins in human colon cancer stroma. The main aim of the present study was to identify novel prognostic markers in human PDAC stroma using our strategies.
Five primary cultivated fibroblasts from human pancreas were stimulated by two types of pancreatic cancer-cell-conditioned medium (Capan-1 and MIA PaCa-2) followed by gene expression analysis to identify up-regulated genes. Publicly available microarray data set concomitant with overall survival was collected and prognostic marker candidates were selected among the genes that were found to be up-regulated. The mRNA expression levels of the selected genes were evaluated in 5 human fresh PDAC tissues. Finally, survival analysis was performed based on immunohistochemical results on tissue microarrays consisting of 216 surgically resected PDAC tissues.
The microarray data of the cancer-stromal interaction model revealed that 188 probes were significantly regulated in pancreatic fibroblasts. Further, six genes were selected using publicly available microarray data set, and a single Diaphanous-related formin-3 (DIAPH3), actin-binding protein, was identified as a stromal biomarker in PDAC fibroblasts by RNA validation analysis. DIAPH3 exhibited strong immunohistochemical expression in stromal fibroblasts. The high stromal expression of DIAPH3 was associated with shorter survival times of PDAC patients.
DIAPH3 was identified as a prognostic marker in PDAC fibroblasts using our biomarker screening strategies through the cancer-stromal interaction model, indicating that stromal actin-binding proteins might have an important biological role in cancer progression. These strategies were also available in PDAC, and can be used for stromal biomarker screening in various cancers.
与胰腺导管腺癌（PDAC）相关的基质成纤维细胞通过与癌细胞相互作用在肿瘤进展中发挥重要作用。我们之前通过癌症-基质相互作用模型提出了体外和计算机筛选生物标志物的联合策略，在人类结肠癌基质中鉴定了几种肌动蛋白结合蛋白。本研究的主要目的是使用我们的策略在人类 PDAC 基质中鉴定新的预后标志物。
用两种胰腺癌细胞条件培养基（Capan-1 和 MIA PaCa-2）刺激 5 个人胰腺原代培养的成纤维细胞，然后进行基因表达分析，以鉴定上调基因。收集了具有总生存数据的公共微阵列数据集，并在发现上调的基因中选择了预后标志物候选物。在 5 个人的新鲜 PDAC 组织中评估了所选基因的 mRNA 表达水平。最后，基于包含 216 例手术切除 PDAC 组织的组织微阵列的免疫组织化学结果进行了生存分析。
癌症-基质相互作用模型的微阵列数据显示，188 个探针在胰腺成纤维细胞中显著调节。此外，使用公共微阵列数据集选择了六个基因，并且通过 RNA 验证分析，发现一个单一的 Diahanous 相关形态发生因子-3（DIAPH3），一种肌动蛋白结合蛋白，是 PDAC 成纤维细胞中的基质生物标志物。DIAPH3 在基质成纤维细胞中表现出强烈的免疫组织化学表达。DIAPH3 在基质中的高表达与 PDAC 患者的生存时间较短相关。
使用我们通过癌症-基质相互作用模型的生物标志物筛选策略，在 PDAC 成纤维细胞中鉴定出 DIAPH3 作为预后标志物，表明基质肌动蛋白结合蛋白在癌症进展中可能具有重要的生物学作用。这些策略也适用于 PDAC，可以用于各种癌症的基质生物标志物筛选。
