乳菇菌素D体外抗肿瘤活性及对细胞周期和凋亡的影响
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摘要
目的:研究来源于绒白乳菇菌的新型倍半萜类化合物——乳菇菌素D的抗肿瘤活性及对细胞周期和细胞凋亡的影响。方法:运用噻唑蓝法检测乳菇菌素D对A549、HCT-8、Bel-7402、SMMC7721、He La细胞增殖的抑制作用并计算半数抑制率(half maximal inhibitory concentration,IC50),并以顺铂作为阳性对照;选择对该化合物敏感的A549细胞株,观察其作用后细胞的形态学改变、细胞凋亡率和细胞周期的变化。结果:乳菇菌素D能够有效抑制A549、Bel-7402、SMMC7721和He La细胞的增殖,作用72 h后的IC50分别为2.46、19.09、18.21、17.05μg/m L;乳菇菌素D可引起A549细胞显著的形态学改变,出现凋亡小体;诱导A549细胞凋亡(100μg/m L作用72 h时的凋亡率为32.14%)、使细胞阻滞于S期。结论:该化合物能够抑制多种组织来源的肿瘤细胞增殖,其中A549细胞最敏感;其作用机制是通过干扰细胞增殖周期而诱导肿瘤细胞凋亡。本实验为乳菇菌素D的开发提供实验依据。
Objective: To investigate the anti-tumor activity of mitissimol D, a novel sesquiterpene compound extracted from Lactarius vellereus, and its effects on cell cycle and apoptosis. Methods: 3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT) assay was used to determine the inhibitory effect of mitissimol D on the proliferation of A549, HCT-8, Bel-7402, SMMC7721 and He La cells and to determine the half maximal inhibition concentration(IC50) in comparison with cisplatin as a positive control. A549 cells sensitive to mitissimol D were selected to observe the morphological changes and the changes in cell apoptosis rate and cell cycle. Results: Mitissimol D could effectively inhibit the proliferation of A549, Bel-7402, SMMC7721 and He La cells with IC50(72 h) of 2.46, 19.09, 18.21 and 17.05 μg/m L, respectively. Mitissimol D could cause significant morphological changes of A549 cells, leading to the formation of apoptosomes and it also could induce apoptosis in A549 cells( 32.14% apoptosis rate at 100 μg/m L after 72 h) and lead to cell block in the S phase. Conclusion: Mitissimol D can significantly inhibit the proliferation of tumor cells from a variety of tissue sources, of which A549 cells are the most sensitive, by interfering with the cell cycle. These findings provide an experimental basis for the development of mitissimol D as a new sesquiterpene compound.
Objective: To investigate the anti-tumor activity of mitissimol D, a novel sesquiterpene compound extracted from Lactarius vellereus, and its effects on cell cycle and apoptosis. Methods: 3-(4,5)-Dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide(MTT) assay was used to determine the inhibitory effect of mitissimol D on the proliferation of A549, HCT-8, Bel-7402, SMMC7721 and He La cells and to determine the half maximal inhibition concentration(IC50) in comparison with cisplatin as a positive control. A549 cells sensitive to mitissimol D were selected to observe the morphological changes and the changes in cell apoptosis rate and cell cycle. Results: Mitissimol D could effectively inhibit the proliferation of A549, Bel-7402, SMMC7721 and He La cells with IC50(72 h) of 2.46, 19.09, 18.21 and 17.05 μg/m L, respectively. Mitissimol D could cause significant morphological changes of A549 cells, leading to the formation of apoptosomes and it also could induce apoptosis in A549 cells( 32.14% apoptosis rate at 100 μg/m L after 72 h) and lead to cell block in the S phase. Conclusion: Mitissimol D can significantly inhibit the proliferation of tumor cells from a variety of tissue sources, of which A549 cells are the most sensitive, by interfering with the cell cycle. These findings provide an experimental basis for the development of mitissimol D as a new sesquiterpene compound.
引文
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[4]贺宇彤,梁迪,李道娟,等.河北省2 0 1 2年恶性肿瘤发病与死亡估计[J].中国肿瘤,2016,25(12):965-972.DOI:10.11735/j.issn.1004-0242.2016.12.A005.
[5]付振涛,徐爱强,鹿子龙,等.山东省2 0 1 2年恶性肿瘤发病与死亡[J].中国肿瘤,2016,25(12):942-949.DOI:10.11735/j.issn.1004-0242.2016.12.A002.
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[24]乔巍,汪小莞,李晓春,等.四种倍半萜类化合物广谱抗肿瘤效应的研究进展[J].中国生化药物杂志,2016,36(3):5-8.DOI:10.3969/j.issn.1005-1678.2016.03.02.
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[29]王佳丽,王秀,夏泉,等.莪术油中3种倍半萜类化合物对肝癌Hep G2细胞增殖抑制作用的研究[J].中成药,2014,36(7):1535-1539.DOI:10.3969/j.issn.1001-1528.2014.07.045.
[30]IMAMURA Y,YUKAWA M,UENO M,et al.3,6-Epidioxy-1,10-bisaboladiene inhibits G1-specific transcription through Swi4/Swi6and Mbp1/Swi6 via the Hog1 stress pathway in yeast[J].FEBS Journal,2014,281:4612-4621.DOI:10.1111/febs.12965.