MicroRNA Profiling Implies New Markers of Gemcitabine Chemoresistance in Mutant p53 Pancreatic Ductal Adenocarcinoma.
MicroRNA Profiling Implies New Markers of Gemcitabine Chemoresistance in Mutant p53 Pancreatic Ductal Adenocarcinoma.
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DOI：
10.1371/journal.pone.0143755
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Haier J
中科院分区：
文献类型：
--
作者：
Dhayat SA;Mardin WA;Seggewiß J;Ströse AJ;Matuszcak C;Hummel R;Senninger N;Mees ST;Haier J
胰腺导管腺癌 (PDAC) 中不存在吉西他滨化疗敏感性的可靠预测因子。 MicroRNA (miR) 是表观遗传基因调节因子，在各种癌症中具有肿瘤抑制或致癌作用。本研究评估了化疗耐药性 PDAC 的特定 miR 表达模式。建立了两种突变型 p53 人 PDAC 细胞系的吉西他滨抗性变体。通过细胞毒性和细胞凋亡测定来分析存活率。通过微阵列研究了 1733 个人类 miR 的表达，并通过 qRT-PCR 进行了验证。对失调 miR 的特定靶基因和蛋白质进行计算机分析后，通过蛋白质印迹定量 MRP-1、Bcl-2、突变体 p53 和 CDK1 的表达。与亲代细胞相比，两个建立的 PDAC 克隆均显示出对吉西他滨的显着耐药性 (p<0.02)，且凋亡率较低 (p<0.001)。 MiR 筛选显示，化疗耐药性 PDAC 中 miR 显着上调（miR-21、miR-99a、miR-100、miR-125b、miR-138、miR-210）和下调 miR（miR-31*、miR-330、miR-378）（p<0.05）。生物信息分析表明这些 miR 参与控制细胞死亡和周期的途径。 MRP-1 (p<0.02) 和 Bcl-2 (p<0.003) 在耐药细胞克隆和突变 p53 (p = 0.023) 的一个克隆中均显着过表达。在吉西他滨耐药的 PDAC 中发现了一致的 miR 表达谱，部分受突变 TP53 基因调节，并具有显着的 MRP-1 和 Bcl-2 过表达。这些结果为进一步阐明PDAC的化疗耐药机制和克服PDAC化疗耐药的治疗方法提供了基础。
No reliable predictors of susceptibility to gemcitabine chemotherapy exist in pancreatic ductal adenocarcinoma (PDAC). MicroRNAs (miR) are epigenetic gene regulators with tumorsuppressive or oncogenic roles in various carcinomas. This study assesses chemoresistant PDAC for its specific miR expression pattern. Gemcitabine-resistant variants of two mutant p53 human PDAC cell lines were established. Survival rates were analyzed by cytotoxicity and apoptosis assays. Expression of 1733 human miRs was investigated by microarray and validated by qRT-PCR. After in-silico analysis of specific target genes and proteins of dysregulated miRs, expression of MRP-1, Bcl-2, mutant p53, and CDK1 was quantified by Western blot. Both established PDAC clones showed a significant resistance to gemcitabine (p<0.02) with low apoptosis rate (p<0.001) vs. parental cells. MiR-screening revealed significantly upregulated (miR-21, miR-99a, miR-100, miR-125b, miR-138, miR-210) and downregulated miRs (miR-31*, miR-330, miR-378) in chemoresistant PDAC (p<0.05). Bioinformatic analysis suggested involvement of these miRs in pathways controlling cell death and cycle. MRP-1 (p<0.02) and Bcl-2 (p<0.003) were significantly overexpressed in both resistant cell clones and mutant p53 (p = 0.023) in one clone. Consistent miR expression profiles, in part regulated by mutant TP53 gene, were identified in gemcitabine-resistant PDAC with significant MRP-1 and Bcl-2 overexpression. These results provide a basis for further elucidation of chemoresistance mechanisms and therapeutic approaches to overcome chemoresistance in PDAC.