五味子素A通过HIF-1α/CXCR4信号通路抑制结直肠癌的生长并诱导细胞凋亡和保护性自噬  

Schizandrin A inhibits the growth of colorectal cancer and induces apoptosis and protective autophagy through the HIF-1α/CXCR4 signaling pathway

作　　者：杨宁波[1] 寇耀[1] 贾映东[1] 王栋 YANG Ning-bo;KOU Yao;JIA Ying-dong;WANG Dong(Department of Gastrointestinal Surgery,Suining Central Hospital,Sichuan 629000,China;Basic Medical College,North Sichuan Medical College,Nanchong,Sichuan 637100,China)

机构地区：[1]遂宁市中心医院胃肠外科,629000 [2]川北医学院基础医学院,637100

出　　处：《现代消化及介入诊疗》2020年第10期1329-1334,共6页Modern Interventional Diagnosis and Treatment in Gastroenterology

基　　金：四川省卫生厅科研课题(130493)。

摘　　要：目的研究五味子素A对结直肠细胞凋亡与自噬的影响,以及对小鼠结直肠癌生长的作用与机制研究。方法通过CCK-8、Annexin V-FITC/PI与Hoechst33342染色检测不同浓度(0、12.5、25、50μmol/L)的五味子素A对小鼠结直肠癌MCA38细胞存活与凋亡的影响;将mRFP-GFP-LC3质粒转染到MCA38细胞,并使用不同浓度(0、12.5、25、50μmol/L)的五味子素A作用后检测细胞自噬情况;荧光定量PCR(qRT-PCR)和蛋白质印迹法(Western Blot)分别检测缺氧诱导因子1α(HIF-1α)、趋化因子受体4(CXCR4)在mRNA与蛋白水平上的表达;将16只雌性裸鼠随机分为对照组和治疗组,每组8只,通过注射MCA38细胞构建结直肠癌小鼠模型,肿瘤长出时开始给药,治疗组小鼠每隔一天腹膜内注射五味子素A(80 mg/kg),对照组小鼠同时腹膜内注射PBS缓冲液,持续4周。在治疗后第7、10、13、16、19、22、25和28天使用游标卡尺测量肿瘤的大小,28d后HE染色观察肿瘤组织。结果与对照组(0μmol/L)比较,不同浓度的五味子素A处理后细胞活力受到抑制(P<0.05),细胞凋亡数目增加(P<0.05),促进了细胞的自噬,下调了细胞中HIF-1α、CXCR4 mRNA与蛋白的表达水平(P<0.05),且均呈剂量依赖性。经过五味子素A治疗后,小鼠肿瘤在治疗16d后体积明显变小(P<0.05),肿瘤病理切片显示肿瘤细胞减少。结论五味子素A能够抑制小鼠结直肠癌的生长,促进结直肠细胞发生凋亡及保护性自噬,其作用可能与抑制HIF-1α/CXCR4信号通路相关。Objective To study the effects of Schizandrin A on colorectal cell apoptosis and autophagy,and its effect and mechanism on colorectal cancer in mice.Methods CCK-8,Annexin V-FITC/PI and Hoechst33342 staining were used to detect the effects of Schizandrin A on the survival and apoptosis of colorectal cancer MCA38 cells in mice(0,12.5,25,50μmol/L);The mRFP-GFP-LC3 plasmid was transfected into MCA38 cells,and schisandrin A at different concentrations(0,12.5,25,50μmol/L)was used to detect cell autophagy;qRT-PCR and Western Blot were used to detect the expression of hypoxia-inducible factor 1α(HIF-1α)and chemokine receptor 4(CXCR4)at the mRNA and protein levels,respectively;Sixteen female nude mice were randomly divided into a control group and a treatment group,with eight in each group,and a mouse model of colorectal cancer was constructed by injecting MCA38 cells;Dosing when tumors grow;The mice in the treatment group were intraperitoneally injected with schisandrin A(80 mg/kg),and the mice in the control group were injected intraperitoneally with PBS buffer for 4 weeks.Use vernier calipers to measure tumor size on days 7,10,13,16,19,22,25 and 28 after treatment,tumor tissue was observed by HE staining after 28 days.Results Compared with the control group(0μmol/L),the cell viability of Schisantherin A at different concentrations was inhibited(P<0.05),and the number of apoptosis increased(P<0.05),which promoted cell autophagy and down-regulated The expression levels of HIF-1α,CXCR4 mRNA and protein in cells(P<0.05)were all dose-dependent.After schisandrin A treatment,the tumor volume in mice became significantly smaller after 16 days of treatment(P<0.05),and tumor pathological sections showed a decrease in tumor cells.Conclusion Schizandrin A can inhibit the growth of colorectal cancer in mice,promote apoptosis and protective autophagy in colorectal cells,and its effect may be related to the inhibition of HIF-1α/CXCR4 signaling pathway.

关 键 词：五味子素A 凋亡 自噬 HIF-1α/CXCR4信号通路 结直肠癌 

分 类 号：R735.34[医药卫生—肿瘤] R574[医药卫生—临床医学]