基于转录组测序探讨白花蛇舌草-半枝莲提取物抗结直肠癌作用机制  

Discussion on Mechanism of Hedyotis Diffusae Herba-Scutellariae Barbatae Herba Extract in Treating Colorectal Cancer Based on Transcriptome Sequencing

作　　者：朱丹烨 吴美美 邓娟 范洪波 洪伟彬 韩织优 ZHU Danye;WU Meimei;DENG Juan;FAN Hongbo;HONG Weibin;HAN Zhiyou

机构地区：[1]广州市东升医院,广东广州510000

出　　处：《新中医》2022年第21期149-155,共7页New Chinese Medicine

基　　金：广州市科技局课题项目(202102080625);广州市民政局课题项目(2020MZK06)。

摘　　要：目的:基于转录组测序探讨白花蛇舌草-半枝莲(HD-SB)提取物对结直肠癌的作用机制。方法:采用不同浓度(0、0.25、0.5、1、2、4 mg/mL)白花蛇舌草(HD)提取物、半枝莲(SB)提取物、HD-SB提取物处理3种不同结直肠癌细胞系(HT-29、HCT-116、SW620)24 h,采用CCK8检测细胞活力。选择未处理的SW620细胞(对照组)和1 mg/mL HD-SB处理24 h的SW620细胞(HD-SB组)进行转录组测序,实时PCR检测靶向无翅型MMTV整合位点家族(WNT)通路5个关键基因[无翅型MMTV整合位点家族成员11(WNT11)、无翅型MMTV整合位点家族成员2B(WNT2B)、卷曲同源物2(FZD2)、低密度脂蛋白受体相关蛋白6(LRP6)、磷脂酶C-β4(PLCβ4)]表达。结果:与HD提取物、SB提取物相比,1、2、4 mg/mL这3个浓度HD-SB提取物处理HT-29、SW620细胞后,活力显著下降(P<0.05)。转录组测序发现,与对照组比较,HD-SB组1522个差异基因发生显著变化(P<0.05),其中有203个基因表达显著上调(P<0.05),1319个基因表达显著下调(P<0.05)。GO富集分析显示,mRNA中的差异基因主要与细胞组分、细胞表达调控、免疫反应的调控等功能相关;KEGG分析显示,差异基因参与的通路主要有代谢相关信号通路、癌症相关通路、丝裂原活化蛋白激酶(MAPK)信号通路、WNT信号通路等。与对照组比较,HD-SB组WNT通路的WNT11、WNT2B、FZD2、LRP6、PLCβ45个关键基因表达均显著下调(P<0.05)。结论:HD-SB提取物可抑制结直肠癌SW620细胞增殖,其机制可能与抑制WNT通路活化有关。Objective:To discuss the mechanism of Hedyotis Diffusae Herba-Scutellariae Barbatae Herba(HD-SB)extract in treating colorectal cancer based on transcriptome sequencing.Methods:Different concentrations(0,0.25,0.5,1,2,4 mg/mL)of Hedyotis Diffusa Herba(HD)extract,Scutellariae Barbatae Herba(SB)extract and HD-SB extract were used for treating three colorectal cancer cell lines(HT-29,HCT-116,SW620)for 24 hours,and the cell viability was detected by CCK8.The untreated SW620cells(the control group)and SW620 cells which were treated with HD-SB at the concentration of 1 mg/mL for 24 hours(the HD-SB group)were selected for transcriptome sequencing.The expression of five key genes,including winglesstype MMTV integration site family member 11(WNT11),winglesstype MMTV integration site family member 2B(WNT2B),frizzled homolog 2(FZD2),low density lipoprotein receptorrelated protein 6(LRP6)and phospholipase C-β4(PLCβ4),targeting winglesstype MMTV integration site family(WNT)pathway was detected by real-time PCR.Results:The HD,SB and HD-SB extracts at the concentration of 2.4 mg/mL can significantly inhibit cell viability of HCT-116,HT-29 and SW620(P<0.05);the viability of HT-29 and SW620 cells treated with HD-SB extract at the concentrations of 1,2 and4 mg/mL was significantly decreased when compared with that treated with HD extract and SB extract(P<0.05).Transcriptome sequencing showed that significant changes were found in the comparisons of 1522 differential genes between the HD-SB group and the control group(P<0.05),of which 203 genes in the HD-SB group were increased(P<0.05)and 1319 genes were decreased(P<0.05).GO functional enrichment analysis showed that the differential genes in mRNA were mainly related to functions of cell components,regulation of cell expression,and regulation of immune responses;KEGG signaling pathway enrichment analysis showed that differential genes were mainly involved in the metabolismrelated signaling pathways,cancer-related pathways,mitogen activated protein kinase(MAPK)signaling pathway,WNT

关 键 词：结直肠癌 半枝莲 白花蛇舌草 混合提取物 细胞增殖 转录组测序 

分 类 号：R735.3[医药卫生—肿瘤]