采用退火控制引物系统鉴定 III 期浆液性卵巢癌中的差异表达基因。
Identification of differentially expressed genes using an annealing control primer system in stage III serous ovarian carcinoma.
机构信息
Department of Obstetrics and Gynecology, Soonchunhyang University Chunan Hospital, Chungcheongnam-do, Korea.
出版信息
BMC Cancer. 2010 Oct 22;10:576. doi: 10.1186/1471-2407-10-576.
BACKGROUND
Most patients with ovarian cancer are diagnosed with advanced stage disease (i.e., stage III-IV), which is associated with a poor prognosis. Differentially expressed genes (DEGs) in stage III serous ovarian carcinoma compared to normal tissue were screened by a new differential display method, the annealing control primer (ACP) system. The potential targets for markers that could be used for diagnosis and prognosis, for stage III serous ovarian cancer, were found by cluster and survival analysis.
METHODS
The ACP-based reverse transcriptase polymerase chain reaction (RT PCR) technique was used to identify DEGs in patients with stage III serous ovarian carcinoma. The DEGs identified by the ACP system were confirmed by quantitative real-time PCR. Cluster analysis was performed on the basis of the expression profile produced by quantitative real-time PCR and survival analysis was carried out by the Kaplan-Meier method and Cox proportional hazards multivariate model; the results of gene expression were compared between chemo-resistant and chemo-sensitive groups.
RESULTS
A total of 114 DEGs were identified by the ACP-based RT PCR technique among patients with stage III serous ovarian carcinoma. The DEGs associated with an apoptosis inhibitory process tended to be up-regulated clones while the DEGs associated with immune response tended to be down-regulated clones. Cluster analysis of the gene expression profile obtained by quantitative real-time PCR revealed two contrasting groups of DEGs. That is, a group of genes including: SSBP1, IFI6 DDT, IFI27, C11orf92, NFKBIA, TNXB, NEAT1 and TFG were up-regulated while another group of genes consisting of: LAMB2, XRCC6, MEF2C, RBM5, FOXP1, NUDCP2, LGALS3, TMEM185A, and C1S were down-regulated in most patients. Survival analysis revealed that the up-regulated genes such as DDAH2, RNase K and TCEAL2 might be associated with a poor prognosis. Furthermore, the prognosis of patients with chemo-resistance was predicted to be very poor when genes such as RNase K, FOXP1, LAMB2 and MRVI1 were up-regulated.
CONCLUSION
The DEGs in patients with stage III serous ovarian cancer were successfully and reliably identified by the ACP-based RT PCR technique. The DEGs identified in this study might help predict the prognosis of patients with stage III serous ovarian cancer as well as suggest targets for the development of new treatment regimens.
背景
大多数卵巢癌患者被诊断为晚期疾病（即 III 期-IV 期），预后不良。通过新的差异显示方法——退火控制引物（ACP）系统，筛选出 III 期浆液性卵巢癌与正常组织相比差异表达的基因（DEGs）。通过聚类和生存分析，发现 III 期浆液性卵巢癌潜在的可用于诊断和预后的标记物的潜在靶点。
方法
采用 ACP 逆转录聚合酶链反应（RT-PCR）技术鉴定 III 期浆液性卵巢癌患者的 DEGs。通过定量实时 PCR 对 ACP 系统鉴定的 DEGs 进行验证。根据定量实时 PCR 表达谱进行聚类分析，采用 Kaplan-Meier 法和 Cox 比例风险多变量模型进行生存分析；比较化疗耐药组和化疗敏感组的基因表达结果。
结果
通过 ACP 逆转录 PCR 技术共鉴定出 114 个 III 期浆液性卵巢癌患者的 DEGs。与凋亡抑制过程相关的 DEGs 倾向于上调克隆，而与免疫反应相关的 DEGs 倾向于下调克隆。定量实时 PCR 获得的基因表达谱聚类分析显示两组截然不同的 DEGs。即一组包括：SSBP1、IFI6、DDT、IFI27、C11orf92、NFKBIA、TNXB、NEAT1 和 TFG 的基因上调，另一组由 LAMB2、XRCC6、MEF2C、RBM5、FOXP1、NUDCP2、LGALS3、TMEM185A 和 C1S 组成的基因下调。生存分析显示，上调基因如 DDAH2、RNase K 和 TCEAL2 可能与预后不良相关。此外，当 RNase K、FOXP1、LAMB2 和 MRVI1 等基因上调时，预测化疗耐药患者的预后极差。
结论
通过 ACP 逆转录 PCR 技术成功可靠地鉴定出 III 期浆液性卵巢癌患者的 DEGs。本研究鉴定的 DEGs 可能有助于预测 III 期浆液性卵巢癌患者的预后，并为新的治疗方案的开发提供潜在靶点。
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