circBFAR调控hsa-miR-424-5p及PI3K-Akt信号通路基因促进胰腺癌进展的机制研究  

Mechanism of circBFAR regulating hsa-miR-424-5p and PI3K-Akt signaling pathway genes to promote the progression of pancreatic cancer

作　　者：依力哈木·买买提[1] 杨欢[2] 再依奴尔·阿不都外力[2] 依马木买买提江·阿布拉[2] Yilihamu·Maimaiti;Yanghuan;Zaiyinuoer·Abulawaili;Yimamumaimaitijiang·Abula(Affiliated Cancer Hospital of Xinjiang Medical University,830054,China;The First Affiliated Hospital of Xinjiang Medical University,830011,China)

机构地区：[1]新疆医科大学第一附属医院日间诊疗中心,830054 [2]新疆医科大学附属肿瘤医院肝胆胰外科,830011

出　　处：《现代消化及介入诊疗》2024年第9期1068-1076,共9页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：新疆维吾尔自治区自然科学基金(2022D01C292)。

摘　　要：目的探讨circBFAR在胰腺癌中的差异表达及其作用分子机制。方法在GSE69362数据集中分析circBFAR在胰腺癌和对照组织中的差异表达。利用TCGA数据库鉴定了胰腺癌和对照组织间的差异表达miRNAs(DEmiRs),并与circBFAR的靶标miRNAs进行交集分析。在GSE62452和GSE28735数据集中鉴定了胰腺癌和对照组织间的差异表达mRNAs(DEmRs),并与靶标DEmiRs的靶向调控mRNAs进行交集分析。对circBFAR靶标DEmiRs调控的DEmRs进行富集分析,筛选出参与PI3K-Akt信号通路的基因并绘制生存曲线。收集10例胰腺癌组织及相应的癌旁对照组织样本,进行RT-qPCR和Western blot检测PI3K-Akt信号通路基因的表达。在胰腺癌细胞系PANC-1中敲降circBFAR或hsa-miR-424-5p,将细胞分为五组进行实验:对照组;si-circNC组;si-circBFAR组;si-circBFAR+inhibitor NC组;si-circBFAR+hsa-miR-424-5p inhibitor组。利用CCK-8检测细胞活性,通过划痕实验和Transwell实验检测细胞迁移和侵袭,利用RT-qPCR和Western blot检测信号通路基因的表达。结果circBFAR在胰腺癌组织中的表达显著高于对照组(P<0.001)。在鉴定的靶标DEmiRs中,hsa-miR-424-5p在胰腺癌中低表达(P<0.01)。hsa-miR-424-5p调控的靶标DEmRs中,LAMA3、ITGA3、MET和AREG参与PI3K-Akt信号通路,在胰腺癌中高表达时患者预后不良(P<0.001)。RT-qPCR和Western blot检测结果显示,circBFAR、LAMA3、ITGA3、MET和AREG在胰腺癌患者中高表达,而hsa-miR-424-5p则低表达。在PANC-1细胞中转染si-circBFAR显著抑制了细胞增殖、迁移和侵袭,以及LAMA3、ITGA3、MET和AREG的表达。转染hsa-miR-424-5p inhibitor抑制了si-circBFAR对细胞的影响(P<0.01)。结论circBFAR通过调控hsa-miR-424-5p及其靶向基因LAMA3、ITGA3、MET和AREG的表达,参与PI3K-Akt信号通路,促进了胰腺癌的进展。抑制circBFAR可能成为胰腺癌治疗的潜在策略。Objective To investigate the differential expression of circBFAR in pancreatic cancer and its molecular mechanisms of action.Methods The differential expression of circBFAR in pancreatic cancer and control tissues was analyzed in the GSE69362 dataset.Differentially expressed miRNAs(DEmiRs)between pancreatic cancer and control tissues were identified using the TCGA database,and an intersection analysis was performed with circBFAR target miRNAs.Differentially expressed mRNAs(DEmRs)between pancreatic cancer and control tissues were identified in the GSE62452 and GSE28735 datasets,and an intersection analysis was conducted with the target-regulated mRNAs of the identified DEmiRs.Enrichment analysis was performed on DEmRs regulated by circBFAR target DEmiRs to identify genes involved in the PI3K-Akt signaling pathway,and survival curves were plotted.Ten pancreatic cancer tissues and corresponding adjacent normal tissues were collected,and RT-qPCR and Western blot were used to detect the expression of PI3K-Akt pathway genes.In the pancreatic cancer cell line PANC-1,circBFAR or hsa-miR-424-5p was knocked down,and the cells were divided into five groups for the experiments:control group,si-circNC group,si-circBFAR group,si-circBFAR+inhibitor NC group,and si-circBFAR+hsa-miR-424-5p inhibitor group.Cell viability was detected using the CCK-8 assay,cell migration and invasion were assessed by wound healing and Transwell assays,and the expression of signaling pathway genes was detected by RT-qPCR and Western blot.Results The expression level of circBFAR in pancreatic cancer tissues was significantly higher than that in the control group(P<0.001).Among the identified target DEmiRs,hsa-miR-424-5p was downregulated in pancreatic cancer(P<0.01).Target DEmRs regulated by hsa-miR-424-5p,including LAMA3,ITGA3,MET,and AREG,are involved in the PI3K-Akt signaling pathway,and their high expression in pancreatic cancer is associated with poor prognosis(P<0.001).RT-qPCR and Western blot results showed that circBFAR,LAMA3,ITGA3,MET,and AR

关 键 词：胰腺癌 circBFAR PI3K-AKT信号通路 hsa-miR-424-5p 

分 类 号：R737.14[医药卫生—肿瘤] R576[医药卫生—临床医学]