喜树碱及其衍生物对肝肿瘤细胞SMMC-7721 增殖的影响

范晓月; 郭 明; 顾 奕; 何云核; 赵富荣

doi:10.3969/j.issn.1001-3776.2020.03.004

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

摘要

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（40 μg·mL-1和80 μg·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。

Abstract: 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 (40 μg/mL and 80 μg/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.

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