喜树碱及其衍生物对肝肿瘤细胞SMMC-7721增殖的影响  

Effect of Camptothecin and its Derivatives on Proliferation of Liver Tumor Cell SMMC-7721

作　　者：范晓月 郭明 顾奕[3] 何云核 赵富荣[3] FAN Xiao-yue;GUO Ming;GU Yi;HE Yun-he;ZHAO Fu-rong(School of Landscape Architecture,Zhejiang A&F University,Hangzhou 311300,China;School of Science,Zhejiang A&F University,Hangzhou 311300,China;School of Forestry and Bio-technology,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学风景园林与建筑学院,浙江杭州311300 [2]浙江农林大学理学院,浙江杭州311300 [3]浙江农林大学林业与生物技术学院,浙江杭州311300

出　　处：《浙江林业科技》2020年第3期22-30,共9页Journal of Zhejiang Forestry Science and Technology

基　　金：国家自然科学基金(31470587)资助;浙江省省院合作科技项目(2015SY11)资助。

摘　　要：采用CCK-8法、荧光定量PCR(RT-PCR)法、蛋白免疫印迹(Western blot)法和活性氧(ROS)测定方法,探讨喜树有效成分喜树碱(CPT)及其衍生物10-羟基喜树碱(HCPT)和盐酸伊立替康(CPT-11)对肝肿瘤细胞SMMC-7721增殖的影响。实验结果表明,SMMC-7721细胞在暴露于CPT-11和HCPT(40μg·mL-1和80μg·mL-1)24 h和48 h后,细胞增殖受到显著抑制(P<0.05)。细胞凋亡检测结果表明,细胞以浓度依赖性方式凋亡。RT-PCR检测结果显示,过氧化氢酶基因(CAT)和超氧化物歧化酶基因(SOD1)的mRNA表达水平显著下降(P<0.05)。Western blot分析证实,CAT和SOD1蛋白表达下调与RT-PCR检测结果一致。ROS检测结果显示,CPT/HCPT/CPT-11显著促进ROS的产生(P<0.05)。所以,CPT/HCPT/CPT-11可下调细胞抗氧化相关基因CAT和SOD1的表达,抑制细胞增殖,显示出抗肿瘤活性,对肝肿瘤细胞的临床治疗有一定的参考价值。Determinations were performed by CCK-8 method,fluorescent quantitative PCR(RT-PCR),Western blot and reactive oxygen species(ROS)on effect of camptothecin(CPT)and its derivatives like 10-hydroxycamptothecin(HCPT)and irinotecan hydrochloride(CPT-11)on proliferation of liver cancer cell SMMC-7721.The result showed that SMMC-7721 were significantly inhibited after 24 and 48 hours of exposure to CPT-11 and HCPT(40μg/mL and 80μg/mL)(P<0.05).Apoptosis detection indicated that SMMC-772 was concentration-dependent.RT-PCR showed that the mRNA expression of catalase gene(CAT)and superoxide dismutase gene(SOD1)decreased significantly(P<0.05).Western blot analysis confirmed that CAT and SOD1 protein expression was down-regulated,the same result of RT-PCR detection.ROS test demonstrated that CPT/HCPT/CPT-11 significantly promoted the generation of ROS(P<0.05).Therefore,CPT/HCPT/CPT-11 could down-regulate the expression of CAT and SOD1,inhibit cell proliferation.

关 键 词：喜树碱 10-羟基喜树碱 盐酸伊立替康 SMMC-7721细胞 增殖抑制 氧化损伤 

分 类 号：TQ351.01[化学工程]