摘要
目的探讨鸟氨酸脱羧酶1(ODC1)基因对前列腺癌细胞PC-3 LncRNA表达的影响,为进一步研究其作用及调控机制提供一些实验基础和线索。方法以前列腺癌细胞PC-3作为研究对象,敲低ODC1基因,经q-PCR检测及WB检测验证后采用Illumina Novaseq 6000,PE150模式进行高通量测序,测序结果应用编码潜能分析方法(CPC分析、CNCI分析、CPAT分析、LGC分析)进行预测筛选,对筛选出的lncRNAs行差异分析、顺式作用元件分析以及GO和KEGG富集分析。再选择文献报道中与前列腺癌关系密切的几个功能lncRNAs,采用q-PCR进行验证。结果(1)在ODC1敲低PC-3细胞中筛选到341个lncRNA,差异表达的lncRNA中上调154个,下调128个。符合条件的差异lncRNA与差异表达基因共表达的顺式调控靶标有38个。(2)差异表达lncRNAs行GO和KEGG富集分析显示,lncRNA靶标基因参与体内多种生物学通路,如磷脂酰肌醇3激酶/蛋白激酶B信号通路(PI3K-Akt信号通路)、Ras信号通路等,还与肿瘤血管生成、细胞群增殖、正调控细胞分裂等密切相关。(3)q-PCR法验证结果:LINC00973、TERC表达显著下调、LINC00638表达显著上调,与GO和KEGG富集分析得到的lncRNA差异表达情况一致。结论敲低ODC1基因可导致PC-3细胞lncRNAs差异表达,ODC1基因有可能通过lncRNAs影响细胞信号通路,促进肿瘤的发生发展。本实验为进一步研究ODC1基因在前列腺癌中的作用提供了一些新的实验基础。
Objective To explore the effect of ornithine decarboxylase 1(ODC1)gene on PC-3 LncRNA expression in prostate cancer cells,and to provide some experimental basis and clues for further study of its role and regulation mechanism.Methods Prostate cancer cell PC-3 was selected as the research object,ODC1 gene was knocked down,q-PCR detection and WB detection were verified and Illumina Novaseq 6000 and PE150 models were used for high-throughput sequencing.Coding potential analysis method(CPC analysis,CNCI analysis,CPAT analysis and LGC analysis)were used to predict and screen the sequencing results,and the selected lncRNAs were analyzed by difference analysis,cis-acting element analysis and GO and KEGG enrichment analysis.Several functional lncRNAs reported to be closely related to prostate cancer were selected and verified by q-PCR.Results (1)341 lncRNA were selected in ODC1 knockdown PC-3 cells,with 154 upregulated and 128 downregulated in the differentially expressed lncRNA.There were 38 cis-regulated targets of eligible differential lncRNA being differentially expressed with differentially expressed genes.(2)GO and KEGG enrichment analysis of differential expressed lncRNAs showed that lncRNA target genes were involved in various biological pathways in vivo,such as phosphatidylinositol 3 kinase/protein kinase B signaling pathway(PI3K-Akt signaling pathway),Ras signaling pathway,and were also closely related to tumor angiogenesis,cell population proliferation,and positive regulation of cell division.(3)q-PCR validation results:LINC00973 and TERC was significantly downregulated,and LINC00638 was significantly upregulated,which was consistent with the differential expression of lncRNA obtained by GO and KEGG enrichment analysis.Conclusion Knockdown of ODC1 leads to differential expression of lncRNAs in PC-3 cells,and it is possible that ODC1 gene affects cell signaling pathway through lncRNAs to promote the occurrence and development of tumors.This experiment provides some new experimental basis for further investigati
作者
宋尖卓
马健
陈鹏
SONG Jianzhuo;MA Jian;CHEN Peng(Department of Urology,the Affiliated Tumor Hospital,Xinjiang Medical University,Urumqi 830011,China)
出处
《新疆医科大学学报》
CAS
2024年第4期526-532,共7页
Journal of Xinjiang Medical University
基金
新疆维吾尔自治区自然科学基金项目(2022D01C285)。
