蛹虫草水提物影响人胃癌细胞BGC-823不同RCD通路的转录组分析  

Transcriptome Analysis of Effect of Cordyceps militaris Aqueous Extract in Human Gastric Cancer Cell BGC-823 on Different RCD Pathways

作　　者：崔明琦 于凯雯 宋扬 徐方旭 王升厚 王泽[1,2,3] CUI Mingqi;YU Kaiwen;SONG Yang;XU Fangxu;WANG Shenghou;WANG Ze(College of Life Sciences of Shenyang Normal University,Shenyang 110034,China;Key Laboratory of Cordyceps militaris with Functional Value of Liaoning Province,Shenyang 110034,China;Experimental Teaching Center,Shenyang Normal University,Shenyang 110034,China)

机构地区：[1]沈阳师范大学生命科学学院,辽宁沈阳110034 [2]辽宁省功能性蛹虫草重点实验室,辽宁沈阳110034 [3]沈阳师范大学实验教学中心,辽宁沈阳110034

出　　处：《食品工业科技》2025年第4期10-18,共9页Science and Technology of Food Industry

基　　金：辽宁省教育厅服务地方项目(LJKFZ202202662);辽宁省科技厅面上项目(2023-MS-251);辽宁省属本科高校基本科研业务费创新团队支持计划专项(2024-132)。

摘　　要：目的:基于转录组测序技术,分析预测蛹虫草(XG)水提物影响人胃癌细胞BGC-823生长活性的主要调节性细胞死亡(Regulated Cell Death,RCD)信号通路。方法:MTT法测定XG水提物对胃癌细胞增殖能力的影响,转录组测序分析显著差异基因及其京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集情况,获得凋亡、自噬、坏死性凋亡及铁死亡信号通路的hub基因(10个)。通过激光共聚焦、流式细胞术、实时荧光定量PCR(qRT-PCR)进一步验证XG水提物对胃癌细胞凋亡的诱导情况。结果:XG水提物对人胃癌细胞BGC-823生长活性的抑制作用具有剂量依赖特征。转录组测序共筛选到5885个差异表达基因,并在凋亡(P=0.001)及自噬(P=0.008)信号通路存在显著性富集,除铁死亡外,其它信号通路的hub基因的表达量均被显著性抑制,包括caspase依赖性凋亡信号通路的关键因子CASP3、CASP9、CASP7和APAF1,自噬小体形成相关的基因ULK1、ULK2、UVRAG、Atg14及Atg5。细胞学检测结果显示,胃癌细胞经XG水提物处理后呈典型的凋亡形态学特征,早期凋亡率为24.8%,细胞周期阻滞于G2/M期。qRT-PCR检测显示,BCL2、BCL2L1、CASP3、CASP9及XIAP的表达受到显著抑制(P<0.01),ENDOG和AIF表达量显著上调(P<0.01),与转录组测序结果一致。结论:蛹虫草主要通过caspase非依赖性线粒体凋亡途径抑制人胃癌细胞BGC-823的生长活性,同时可以影响自噬等信号通路的激活,为蛹虫草在功能性食品的开发提供理论支持。Objective:The main regulated cell death(RCD)signaling pathway of Cordyceps militaris(XG)water extract that affects the growth activity of the human gastric cancer cell line BGC-823 was analyzed and predicted using transcriptome sequencing technology.Methods:The effect of XG aqueous extract on the proliferation of gastric cancer cells was determined using the MTT method.Transcriptome analysis revealed significant differentially expressed genes(DEGs)and their Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment,resulting in the identification of 10 hub genes involved in the apoptosis,autophagy,necroptosis,and ferroptosis signaling pathways.Laser confocal microscopy,flow cytometry,and quantitative real time polymerase chain reaction(qRT-PCR)were employed to further validate the induction of apoptosis in gastric cancer cells by the XG aqueous extract.Results:The inhibitory effect of XG water extract on the growth of the human gastric cancer cell line BGC-823 was dose-dependent.A total of 5885 differentially expressed genes were identified through transcriptome sequencing analysis,with significant enrichment in the apoptosis(P=0.001)and autophagy(P=0.008)signaling pathways.In addition to ferroptosis,the expression levels of hub genes associated with other signaling pathways were significantly inhibited,including the key factor of caspase-dependent apoptosis(CASP3,CASP9,CASP7,APAF1)as well as ULK1,ULK2,UVRAG,Atg14,and Atg5 related to autophagosome formation.Cytological examination revealed typical apoptotic morphological characteristics in gastric cancer cells treated with the XG water extract,with an early apoptotic rate reaching 24.8%,while cell cycle arrest occurred at the G2/M stage.qRT-PCR showed that the expression of BCL2,BCL2L1,CASP3,CASP9,and XIAP was significantly inhibited(P<0.01),whereas that of ENDOG and AIF was significantly upregulated(P<0.01),consistent with the results of transcriptome sequencing.Conclusion:C.militaris primarily inhibits the growth of the human gastric cancer cell line BGC-823 throu

关 键 词：蛹虫草 凋亡 自噬 坏死性凋亡 铁死亡 

分 类 号：Q28[生物学—细胞生物学]