前列腺癌和癌旁组织差异表达基因的筛选和鉴定

Screening and identification of differentially expressed genes in prostate cancer and adjacent tissues

目的探讨前列腺癌和癌旁组织差异表达基因和蛋白的筛选和鉴定。方法选取2019年6月至2022年6月我院收集的10例前列腺癌组织和对应癌旁组织作为研究对象,采用运用Illumina Hiseq 2500测序平台完成转录组测序,用定量聚合酶链式反应(PCR)和蛋白质免疫印迹对筛选到的前10位差异表达基因和蛋白进行验证和富集,对两组样本的检测数据进行统计学对比分析。结果与癌旁组织比较,前列腺癌组织中312个差异表达基因,其中上调表达基因201个,下调表达基因111个。其中上调表达显著性最高的5个基因分别是ASPM、CCNB2、CDC20、CENPF、CEP55,下调表达显著性最高的5个基因分别是SLC9A9、SDPR、GFRA1、BMP5、SMOC2。生物学过程分析显示这些差异表达蛋白主要在细胞成分形态发生、细胞内受体介导信号通路和葡萄糖代谢发挥作用。分子功能主要包括结构分子生物活性和氨基酸、脂质和糖代谢酶活性。KEGG通路主要聚类在YAP信号通路、p53信号通路、Wnt信号通路、糖酵解通路和RNA代谢通路。癌旁组织ASPM、CCNB2、CDC20、CENPF、CEP55 mRNA表达水平(1.01±0.12、1.02±0.15、1.07±0.14、1.04±0.13、1.07±0.17)明显低于前列腺癌组织(3.01±0.19、2.56±0.22、2.43±0.27、2.15±0.37、1.99±0.26),差异有统计学意义(t=28.350、17.810、14.170、9.248、9.076,P<0.05)。癌旁组织SLC9A9、SDPR、GFRA1、BMP5和SMOC2 mRNA表达水平(1.01±0.10、1.06±0.08、1.09±0.10、1.05±0.09、1.09±0.09)明显高于前列腺癌组织(0.32±0.11、0.40±0.07、0.58±0.05、0.66±0.06、0.73±0.09),差异有统计学意义(t=14.310、18.600、14.490、11.560、8.917,P<0.05)。癌旁组织ASPM、CCNB2、CDC20、CENPF、CEP55蛋白相对表达水平(1.33±0.12、1.16±0.10、1.10±0.06、1.21±0.09、1.18±0.08)明显低于前列腺癌组织(3.13±0.18、2.74±0.22、2.27±0.19、2.07±0.12、1.78±0.20),差异有统计学意义(t=26.780、20.200、18.700、17.810、8.750,P<0.05)。癌旁组织SLC9A9、SDPR、GFRA1、BMP5和SMOC2蛋白相对表达水平(1.42±0.13、1.38±0.10、1.39±0.08、1.52±0.12、1.36±0.14)明显高于前列腺癌组织(0.30±0.05、0.43±0.05、0.57±0.03、0.76±0.09、0.81±0.14),差异有统计学意义(t=25.390、27.320、31.190、15.710、9.264,P<0.05)。结论前列腺组织表达的基因主要富集于YAP、P53和Wnt信号通路,这些信号通路可能参与了调控前列腺癌的发生和发展。

Objective To explore the screening and identification of differentially expressed genes and proteins in prostate cancer and adjacent tissues.Methods Ten cases of prostate cancer tissue and corresponding adjacent tissues collected by our hospital from June 2019 to June 2022 were selected as the research subjects.The transcriptome sequencing was completed using the Illumina Hiseq 2500 sequencing platform,and the top 10 differentially expressed genes and proteins screened were validated and enriched using quantitative PCR and protein immunoblotting.We conducted statistical analysis on the detection data of two sets of samples.Results Compared with adjacent tissues,there were 312 differentially expressed genes in prostate cancer tissue,of which 201 were upregulated gene and 111 were downregulated gene.The five genes with the highest upregulation significance were ASPM,CCNB2,CDC20,CENPF,CEP55,and the five genes with the highest downregulation significance were SLC9A9,SDPR,GFRA1,BMP5,and SMOC2.Biological process analysis shows that these differentially expressed proteins mainly play a role in cellular component morphogenesis,intracellular receptor mediated signaling pathways,and glucose metabolism.Molecular functions mainly include structural molecular biological activity and amino acid,lipid,and glucose metabolism enzyme activity.The KEGG pathway is mainly clustered in the YAP signaling pathway,p53 signaling pathway,Wnt signaling pathway,glycolysis pathway,and RNA metabolism pathway.The mRNA expression levels of ASPM,CCNB2,CDC20,CENPF,and CEP55 in paracancerous tissues(1.01±0.12,1.02±0.15,1.07±0.14,1.04±0.13,1.07±0.17)were significantly lower than those in prostate cancer tissues(3.01±0.19,2.56±0.22,2.43±0.27,2.15±0.37,1.99±0.26,t=28.350,17.810,14.170,9.248,9.076,P<0.05).The mRNA expression levels of SLC9A9,SDPR,GFRA1,BMP5,and SMOC2 in adjacent cancer tissues(1.01±0.10,1.06±0.08,1.09±0.10,1.05±0.09,1.09±0.09)were significantly higher than those in prostate cancer tissues(0.32±0.11,0.40±0.07,0.58±0.05,0.66±0.06,0.73±0.09,t=14.310,18.600,14.490,11.560,8.917,P<0.05).The relative expression levels of ASPM,CCNB2,CDC20,CENPF,and CEP55 proteins in adjacent tissues(1.33±0.12,1.16±0.10,1.10±0.06,1.21±0.09,1.18±0.08)were significantly lower than those in prostate cancer tissues(3.13±0.18,2.74±0.22,2.27±0.19,2.07±0.12,1.78±0.20,t=26.780,20.200,18.700,17.810,8.750,P<0.05).The relative expression levels of SLC9A9,SDPR,GFRA1,BMP5,and SMOC2 proteins in adjacent cancer tissues(1.42±0.13,1.38±0.10,1.39±0.08,1.52±0.12,1.36±0.14)were significantly higher than those in prostate cancer tissues(0.30±0.05,0.43±0.05,0.57±0.03,0.76±0.09,0.81±0.14,t=25.390,27.320,31.190,15.710,9.264,P<0.05).Conclusion The genes expressed in prostate tissue are mainly enriched in the YAP,P53,and Wnt signaling pathways,which may be involved in regulating the occurrence and development of prostate cancer.