姜黄素对肝癌细胞miR-29及VEGF表达的干预作用  

Interventional Effect of Curcumin on the Expression of miR-29 and VEGF in Liver Cancer Cells

作　　者：陈彩萍 徐琦[3] 赵海菲[2] CHEN Caiping;XU Qi;ZHAO Haifei(Department of Pharmacy,Hangzhou Cancer Hospital,Hangzhou,Zhejiang province,310002,China;Department of Abdominal Oncology,Zhejiang Cancer Hospital,Hangzhou,Zhejiang province,310022,China;Department of Pathology,Hangzhou Cancer Hospital,Hangzhou,Zhejiang province,310002,China)

机构地区：[1]杭州市肿瘤医院药剂科,杭州310002 [2]杭州市肿瘤医院病理科,杭州310002 [3]浙江省肿瘤医院腹部肿瘤内科,杭州310022

出　　处：《浙江中西医结合杂志》2020年第10期785-790,I0001,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的观察姜黄素对肝癌细胞miRNA-29(miR-29)及血管内皮生长因子(VEGF)表达的干预作用。方法收集2008年12月1日—2018年12月1日杭州市肿瘤医院临床资料较完整的手术切除肝癌标本41份,含癌组织和癌旁非癌组织,采用免疫组化研究VEGF表达情况以及与肝癌分型的相关性;转染VEGF siRNA后,MTT检测其对细胞增殖的调控作用,Western blot检测其对凋亡蛋白Bcl2、Bax表达的影响;PCR检测肝癌细胞miR-29表达,转染miR-20mimics后,qRT-PCT和Western blot检测对VEGF表达的影响;MTT检测姜黄素对HepG2细胞抑制效应的IC50,qRT-PCT和Western blot检测姜黄素对miR-29-VEGF调控作用。结果肝癌组织VEGF表达阳性率70.73%(29/41)显著高于癌旁肝组织30.00%(3/10)和正常肝组织(0.00%)(P均<0.01);肝癌组织VEGF表达水平与肿瘤的病理分级、侵袭转移性有关,病理分级Ⅲ~Ⅳ级明显高于Ⅰ~Ⅱ级(P<0.05),高侵袭转移(15/24)明显高于低侵袭转移(7/17)(P<0.05);qRT-PCR结果显示,与转染前比较,转染VEGF siRNA 48h后,细胞增殖下降[(2.31±0.39)%比(4.21±0.52)%,P<0.05];与阴性对照组比较,抑制VEGF表达后,HepG2细胞迁移能力明显降低;与正常人肝细胞比较,HepG2细胞miR-29表达量显著降低[(2.11±0.92)比(7.32±2.11),P<0.05];转染miR-29 mimics促进miR-29表达后,VEGF mRNA表达水平下降[(3.01±1.00)比(11.33±2.02),P<0.05];姜黄素对HepG2细胞抑制率在72h最好(IC50=49.50μmol/L);肝癌HepG2细胞与姜黄素59.68μmol/L共同培养72h后,miR-29表达显著升高[(9.04±2.19)比(2.18±1.00),P<0.05],VEGF mRNA表达降低[(2.98±1.00)比(8.32±1.80),P<0.05]。结论姜黄素可能通过促进肝癌细胞miR-29表达,抑制VEGF表达,调控肝癌细胞生物学过程。Objective To observe the intervention effect of curcumin on the expression of miRNA-29(miR-29)and vascular endothelial growth factor(VEGF)in liver cancer cells.Methods A total of 41 liver cancer specimens including cancerous tissues and adjacent non-cancerous tissues with complete clinical data from December 1,2008 to December 1,2018 in Hangzhou Cancer Hospital were collected.Expression of VEGF and its correlation with liver cancer classification were studied by immunohistochemistry.After transfection with VEGF siRNA,the regulatory effect of VEGF on cell proliferation was detected by MTT,and Western blot was used to test the expression of apoptotic proteins Bcl2 and Bax.Expression of miR-29 was detected by PCR.VEGF expression was checked by qRT-PCR and Western blot after transfection with miR-20Mimics.MTT was used to detect IC50 of curcumin's inhibitory effect on HepG2 cell,qRT-PCR and Western blot were used to assay the regulatory effect of curcumin on miR-29-VEGF.Results The positive rate of VEGF expression in liver cancer tissues was 70.73%(29/41),which was significantly higher than that in adjacent liver tissues 30.00%(3/10)and normal liver tissues(0.00%)(P<0.01).The expression of VEGF was related to the pathological grade and invasion and metastasis of liver cancer tissues.The expression of VEGF in grade Ⅲ-Ⅳ was significantly higher than that in grade Ⅰ-Ⅱ(P<0.05),and that in high invasion(15/24)was significantly higher than that in low invasion(7/17)(P<0.05).The results of qRT-PCR showed that after transfection with VEGF siRNA for 48h,the proliferation of HepG2 cells decreased[(2.31±0.39)%vs(4.21±0.52)%,P<0.05].Compared to the negative control group,the migration ability of HepG2 cells decreased significantly after VEGF siRNA transfection;Compared to normal human hepatocytes,the expression of miR-29 in HepG2 cells decreased significantly[(2.11±0.92)vs(7.32±2.11),P<0.05].After miR-29 mimics were transfected,VEGF mRNA expression decreased[(3.01±1.00)vs(11.33±2.02),P<0.05].Curcumin had the best in

关 键 词：肝癌细胞 姜黄素 MIR-29 VEGF 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]