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| 重楼皂苷Ⅱ联合喜树碱对肺癌H460、H446细胞凋亡及信号通路的影响 |
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郭慧敏1, 李祎亮2, 刘振3
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1.天津技术产权交易有限公司, 天津 300203;2.中国医学科学院北京协和医学院放射医学研究所, 天津 300192;3.中国食品营养/安全与药物化学国际科技合作基地, 教育部工业微生物重点实验室, 天津市工业微生物重点实验室, 天津科技大学生物工程学院, 天津 300457
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| 摘要: |
| [目的]观察重楼皂苷Ⅱ（PSⅡ）联合化疗药物喜树碱（CPT）对肺癌细胞的作用及机制。[方法]采用噻唑蓝（MTT）法检测PSⅡ联合CPT对非小细胞肺癌（NSCLC）细胞（H460）以及小细胞肺癌（SCLC）细胞（H446）的抑瘤效果。平板克隆法观察PSⅡ联合CPT作用于肺癌细胞后的细胞克隆形成率；流式细胞仪检测细胞凋亡情况。蛋白免疫印迹法（Western blot）检测PSⅡ联合CPT对肺癌细胞蛋白激酶B（AKT）、细胞外信号调节酶（ERK）、p38 MAPK的磷酸化活性及抗凋亡蛋白B淋巴细胞瘤（Bcl）-2、Bcl-XL的表达。[结果]MTT实验结果显示PSⅡ联合CPT对肺癌细胞有细胞毒活性并呈现剂量依赖性，且在细胞克隆实验中观察到两者具有很强的协同作用；两者联合增加了H460、H446细胞的晚期凋亡率，且显著增加了H446的早期凋亡率（70.10±3.44）%。Western blot结果显示，PSⅡ联合CPT作用于H460细胞，能明显上调p38 MAPK和ERK的磷酸化蛋白表达，对AKT磷酸化无明显作用，并下调了Bcl-2和Bcl-XL蛋白的表达。PSⅡ联合CPT作用于H446细胞，上调了AKT、p38 MAPK和ERK的磷酸化蛋白表达，并下调了Bcl-2蛋白的表达，对Bcl-XL无明显作用。[结论]PSⅡ可以作为CPT的增敏剂用于肺癌的治疗，为PSⅡ用于肺癌的治疗提供了依据。 |
| 关键词: 重楼皂苷Ⅱ 肺癌 喜树碱 联合用药 |
| DOI：10.11656/j.issn.1672-1519.2019.02.19 |
| 分类号: |
| 基金项目:国家自然科学基金青年项目基金（81703014）。 |
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| Impacts of combination of Paris Saponins Ⅱ and camptothecin on apoptosis and signal pathways in lung cancer H460, H446 cells |
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GUO Huimin1, LI Yiliang2, LIU Zhen3
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1.Tianjin Technology Property Rights Exchange Co., Ltd, Tianjin 300203, China;2.Institute of Radiation Medicine, Chinese Academy of Medical Science and PekingUnion Medical College, Tianjin 300192, China;3.China International Science and Technology Cooperation Base of Food Nutrition/Safety and Medicinal Chemistry, Key Laboratory of Industrial Fermentation Microbiology of Ministry of Education, Tianjin Key Laboratory of Industry Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China
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| Abstract: |
| [Objective] To investigate the effects and mechanism of Pariss saponins (PSⅡ) combined with Camptothecin (CPT) on lung cancer cell line.[Methods] The effects of PSⅡ combined with CPT on the proliferation activity on non-small cell lung cancer H460 cells and small cell lung cancer H446 cells were deteceted by MTT. Cloning plating efficiency of PSⅡ combined with CPT was studied by flat plate clone formation test. Cell apoptosis was analyzed by flow cytometry. Phosphorylation acitivities (AKT, ERK and p38 MAPK) and expression levels of anti-apoptosis protein and Bcl-2 and Bcl-XL were analyzed by Western Blot.[Results] MTT assay showed that PSⅡ combined with CPT had a cytotoxic effect on lung cancer cell in a dose-dependent manner. Strong synergism of PSⅡ combined with CPT was observed in cell clone formation assay. The late apoptosis rate was increased when PSⅡ combined with CPT, and the early apoptosis rate of H446 was significantly increased to (70.10±3.44)%. Western blot result showed that the expression of p38 MAPK and ERK phosphorylation were significantly increased when using PSⅡ combined with CPT on H460, however, there was no obvious effect on the expression of AKT; and the expression of Bcl-2 and Bcl-XL were reduced. PSⅡ combined with CPT can increased the expression of AKT, p38 MAPK and ERK phosphorylation and decreased the expression of Bcl-2 of H446, there was no obvious effect on the expression of Bcl-XL.[Conclusion] PSⅡ exhibits cytotoxicity in different pathways in lung cancer cells and it can be used as a chemosensitizer to CPT in lung cancer cells. |
| Key words: Paris Saponins Ⅱ lung cancer camptothecin combination drugs |
