Hu Zuowei, Zhang Xiaoye, Yang Hang, Qin Shuanglai, Liu Yaqi, Xiong Wei, Yuan Bing, Li Liping, Yao Weiqi, Wu Dongcheng
Department of Oncology, Wuhan Integrated TCM and Western Medicine Hospital, Wuhan, Hubei 430022, P.R. China.
Department of Biochemistry and Molecular Biology, Wuhan University, Wuhan, Hubei 430072, P.R. China.
Oncol Lett. 2017 Dec;14(6):7917-7922. doi: 10.3892/ol.2017.7205. Epub 2017 Oct 18.
The present study aimed at investigating the underlying molecular mechanisms for patients following cytokine-induced killer (CIK) therapy, particularly involving the alterations in p53-associated signaling pathways, to elucidate whether CIK therapy serves a function in cancer treatment. Samples of blood were collected from patients with breast cancer prior to and following CIK therapy. Two group samples were used for RNA sequencing (RNA-Seq) to determine the alterations in gene expression levels following CIK therapy and one for the quantitative polymerase chain reaction (qPCR), to analyze the reliability of RNA-Seq results. The genes that may encode proteins associated with p53 pathways were selected and analyzed. The expression levels of 8 genes were analyzed, including tumor suppressor protein 53 (TP53), murine double minute homolog 2 (MDM2), ribosomal protein L11 (RPL11), ribosomal protein S23 (RPS23), sirtuin 1, histone deacetylase 1, tuberous sclerosis complex 1 (TSC1) and mechanistic target of rapamycin (mTOR), and alterations in expression levels following CIK therapy were determined. However, only RPL11 and RPS23 were identified to exhibit marked alterations in expression levels (FDR <0.05), which was considered to be due to individual distinctions. qPCR analysis revealed that the expression levels of the RPL11, TP53 and TSC1 genes were downregulated, and those of the RPS23 and MDM2 genes were upregulated following CIK therapy. Only MDM2 exhibited a marked alteration in the gene expression level following CIK therapy. Alterations in the expression levels of TP53, RPL11 and TSC1 were associated with those of MDM2, RPS23 and mTOR, respectively.
本研究旨在探究细胞因子诱导的杀伤细胞（CIK）治疗后患者潜在的分子机制，尤其涉及p53相关信号通路的改变，以阐明CIK治疗在癌症治疗中是否发挥作用。在CIK治疗前和治疗后，采集乳腺癌患者的血液样本。两组样本用于RNA测序（RNA-Seq）以确定CIK治疗后基因表达水平的变化，一组用于定量聚合酶链反应（qPCR），以分析RNA-Seq结果的可靠性。选择并分析可能编码与p53通路相关蛋白质的基因。分析了8个基因的表达水平，包括肿瘤抑制蛋白53（TP53）、小鼠双微体同源物2（MDM2）、核糖体蛋白L11（RPL11）、核糖体蛋白S23（RPS23）、沉默调节蛋白1、组蛋白去乙酰化酶1、结节性硬化复合物1（TSC1）和雷帕霉素靶蛋白（mTOR），并确定了CIK治疗后表达水平的变化。然而，仅RPL11和RPS23被鉴定为表达水平有显著变化（FDR<0.05），这被认为是由于个体差异所致。qPCR分析显示，CIK治疗后RPL11、TP53和TSC1基因的表达水平下调，RPS23和MDM2基因的表达水平上调。CIK治疗后，只有MDM2基因表达水平有显著变化。TP53、RPL11和TSC1表达水平的变化分别与MDM2、RPS23和mTOR的变化相关。
