病毒复制是病毒在宿主体内生存、繁殖的关键节点,对人体健康危害极大.体液免疫和细胞免疫是机体调控病毒复制的主要手段,但由于人体免疫系统的复杂性和多样性,仍有大部分病毒复制机制尚未明确.环状RNA是一类具有基因表达调控功能的非编码RNA,通过调节免疫应答因子、miRNA、蛋白质、复制起始点、DNA合成和组装等过程,调控病毒复制.虽然存在环状RNA表达量低、测序建库费用高等问题,但环状RNA仍不失为一个阐述病毒复制机制的好方法.

目的:探究hsa_circ_0078607在结直肠癌组织和患者血清中的表达水平及其与结直肠癌患者临床病理特征的关系,评价其能否作为结直肠癌潜在的分子诊断标志物及治疗靶标.方法:收集2018年6月至2022年1月于柳州市人民医院胃肠外科接受结直肠癌切除手术患者的58对癌及癌旁组织标本,收集2020年1月至2022年12月于柳州市人民医院初次确诊的结直肠癌患者、结直肠息肉患者及健康人体检血清共152例;从结直肠癌差异表达circRNA谱中挑选特异性高表达的hsa_circ_0078607作为候选标志物,采用qPCR法检测其在结直肠癌细胞、组织、患者血清及结直肠息肉患者血清中的相对表达量,分析其与临床病理特征的关系.采用ROC曲线评估hsa_circ_0078607对结直肠癌及结直肠息肉的诊断价值.通过Circular RNA Interactome数据库预测与hsa_circ_0078607结合的miRNA,并用Cytoscape 3.9.1软件构建circRNA-miRNA-mRNA调控网络,同时通过GO/KEGG富集分析进一步了解其功能.结果:与癌旁组织或健康人血清相比,hsa_circ_0078607在结直肠癌细胞、组织和血清及息肉患者血清中呈高表达(P<0.001),其中有52例(89.7%)患者癌组织中表达上调,6例(10.3%)表达下调.结直肠癌组织中hsa_circ_0078607的相对表达量与肿瘤位置(P=0.029)、分化程度(P=0.046)和远处转移(P=0.043)有关联.ROC结果显示,在结直肠癌组织和血清中其诊断结直肠癌的AUC分别为0.845 7[95%CI(0.772 8,0.918 6),P<0.000 1]和0.868 3[95%CI(0.790 7,0.945 9),P<0.000 1];在息肉患者血清中,hsa_circ_0078607诊断结直肠息肉的AUC为0.710 1[95%CI(0.610 0,0.810 1)].GO/KEGG富集分析结果表明,hsa_circ_0078607下游的miRNA可能参与RNA聚合酶Ⅱ启动子转录调控、蛋白K48-连锁泛素化、Wnt、Hippo及MAPK信号通路调控等多个生物过程.结论:Hsa_circ_0078607在结直肠癌细胞、组织和血清中呈高表达,其在结直肠癌组织中的表达水平与肿瘤位置、分化程度和远处转移有关联,提示其可作为结直肠癌潜在的分子诊断标志物;其还可能介导结直肠癌的发生发展过程,对发现结直肠癌潜在的治疗靶点有重要意义.

目的:探讨环状RNA hsa_circ_0006867在结直肠癌中的表达及其与临床病理因素的关系.方法:全转录组测序筛选结直肠癌中特异circRNAs表达谱,挑选出差异表达显著的hsa_circ_0006867,qRT-PCR检测54例结直肠癌组织及癌旁组织中hsa_circ_0006867表达情况,分析其表达水平与结直肠癌临床病理特征的相关性,ROC曲线分析hsa_circ_0006867在结直肠癌中的诊断价值.结果:测序获得circRNAs在结直肠癌中的差异表达谱,qRT-PCR验证hsa_circ_0006867在结直肠癌中表达下调(P<0.05).其表达水平与肿瘤分化程度和远处转移有关(P<0.05).ROC曲线显示hsa_circ_0006867诊断结直肠癌AUC为0.851(95％CI:0.775～0.927),当截断值为0.0146时,敏感度为88.46％(95％CI:0.770～0.946),特异度为73.08％(95％CI:0.598～0.832),差异具有统计学意义(P<0.001).结论:hsa_circ_0006867在结直肠癌中表达下调,与相关临床病理特征密切联系,可作为潜在结直肠癌临床诊断指标.

Background:Gastric cancer (GC) is a primary cause of cancer death around the world. Previous studies have found that Drosha plays a significant role in the development of tumor cells. Soon after, we unexpectedly found that the expression of microRNA6778-5p (miR6778-5p) is unconventionally high in the gastric cancer cells low-expressing Drosha. So, we designed the Drosha interference sequence and recombined it into a lentiviral vector to construct Drosha knockdown lentivirus and transfected the Drosha knockdown lentivirus into gastric cancer cells to establish Drosha knockdown gastric cancer cell lines. We aimed to explore the effect of microRNA6778-5p on the proliferation of gastric cancer cells with Drosha knockdown and its intrinsic mechanism.Methods:We designed the Drosha interference sequence and recombined it into a lentiviral vector to construct Drosha knockdown lentivirus and transfected the Drosha knockdown lentivirus into gastric cancer cells to establish Drosha knockdown gastric cancer cell lines. After transfecting miR6778-5p mimics and inhibitor into gastric cancer cell lines with Drosha knockdown, the expression levels of miR6778-5p mimics in Drosha low-expressing gastric cancer cells increased, while miR6778-5p inhibitor decreased the expression levels of miR6778-5p. The Cell Counting Kit-8 (CCK-8) experiment was used to detect the proliferation ability of gastric cancer cells after overexpression or knockdown of miR6778-5p and bioinformatics predicted the relationship between miR6778-5p and glycogen synthase kinase-3β (GSK3β).Results:After infection with the Drosha knockdown lentivirus, Drosha's mRNA and protein levels were significantly downregulated in gastric cancer cells. The expression levels of miR6778-5p mimics in Drosha low-expressing gastric cancer cells increased, while miR6778-5p inhibitor decreased the expression levels of miR6778-5p. Overexpression of miR6778-5p significantly enhanced the proliferation ability of Drosha low-expression gastric cancer cells; on the contrary, knocking down miR6778-5p weakened the proliferation ability of Drosha low-expression gastric cancer cells. Bioinformatics predicted that miR6778-5p targeted glycogen synthase kinase-3β (GSK3β) and the mRNA and protein levels of GSK3β decreased significantly after overexpression of miR6778-5p.Conclusion:miR6778-5p promotes the proliferation of Drosha low-expressing gastric cancer cells by targeting GSK3β.