研究云芝氯仿提取物对人结肠直肠癌SW - 480细胞的迁移抑制和凋亡作用。
Investigating migration inhibition and apoptotic effects of Fomitopsis pinicola chloroform extract on human colorectal cancer SW-480 cells.
作者信息
Wang Yaqin, Cheng Xiaoxia, Wang Pan, Wang Lu, Fan Jianping, Wang Xiaobing, Liu Quanhong
机构信息
Key Laboratory of Medicinal Resources and Natural Pharmaceutical Chemistry, Ministry of Education, National Engineering Laboratory for Resource Developing of Endangered Chinese Crude Drugs in Northwest of China, College of Life Sciences, Shaanxi Normal University, Xi'an, Shannxi, China.
School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi'an, Shaanxi, China.
出版信息
PLoS One. 2014 Jul 3;9(7):e101303. doi: 10.1371/journal.pone.0101303. eCollection 2014.
BACKGROUND
Fomitopsis pinicola (Sw. Ex Fr.m) Karst (FPK) which belongs to the Basidiomycota fungal class is one of the most popular medical fungi in China. It has been used for many diseases: cancer, heart diseases, diabetes and so on. However, little study on the pro-apoptotic effect and migration inhibition of FPK chloroform extract (FPKc) has been reported and the possible involved mechanism has not been illuminated.
METHODOLOGY/PRINCIPAL FINDINGS: Chemical analysis was performed by HPLC which showed ergosterol (ES) concentration was 105 µg/mg. MTT assay revealed that FPKc could selectively inhibit SW-480 cells viability with the IC50 of 190.28 µg/ml. Wound healing and transwell assay indicated that FPKc could inhibit the migration of SW-480 cells obviously, FPKc could also dramatically decreased the matrix metalloproteinases-2, 9 (MMP-2 and MMP-9) expression. Annexin V-FITC/PI staining, nuclear Hoechst 33342 staining and DNA fragmentation analysis revealed that FPKc and ES could induce SW-480 cells apoptosis. The apoptosis process closely involved in ROS accumulation and depletion of GSH, activation of caspase 3, poly (ADP-ribose) polymerase (PARP) degradation. FPKc could also up-regulate P53 expression and thus lead to G1 phase arrest. When SW-480 cells were pretreated with N-acetylcysteine (NAC), the ROS generation, cell viability and apoptotic ratio were partially declined, which indicated that ROS was vertical in the pro-apoptosis process induced by FPKc. Moreover, in the whole process, ES which has been previously found in FPKc had the similar effect to FPKc. Thus we could conclude that ES, as one of the highest abundant components in FPKc, might also be one of the active constituents.
CONCLUSION/SIGNIFICANCE: FPKc could inhibit the migration of SW-480 cells, induce SW-480 cells G1 phase arrest and cause ROS-mediated apoptosis effect. And ES might be one of the effective constituents in the whole process.
背景
属于担子菌纲真菌类的云杉拟层孔菌（Fomitopsis pinicola (Sw. Ex Fr.m) Karst，FPK）是中国最受欢迎的药用真菌之一。它已被用于治疗多种疾病：癌症、心脏病、糖尿病等。然而，关于云杉拟层孔菌氯仿提取物（FPKc）的促凋亡作用和迁移抑制作用的研究报道较少，其可能涉及的机制也尚未阐明。
方法/主要发现：通过高效液相色谱法进行化学分析，结果显示麦角甾醇（ES）浓度为105μg/mg。MTT法显示FPKc可选择性抑制SW - 480细胞活力，IC50为190.28μg/ml。伤口愈合实验和Transwell实验表明，FPKc可显著抑制SW - 480细胞的迁移，还可显著降低基质金属蛋白酶 - 2、9（MMP - 2和MMP - 9）的表达。膜联蛋白V - FITC/PI染色、细胞核Hoechst 33342染色和DNA片段化分析表明，FPKc和ES可诱导SW - 480细胞凋亡。凋亡过程与活性氧（ROS）积累、谷胱甘肽（GSH）耗竭、半胱天冬酶3激活、聚（ADP - 核糖）聚合酶（PARP）降解密切相关。FPKc还可上调P53表达，从而导致G1期阻滞。当用N - 乙酰半胱氨酸（NAC）预处理SW - 480细胞时，ROS生成、细胞活力和凋亡率部分下降，这表明ROS在FPKc诱导的促凋亡过程中起关键作用。此外，在整个过程中，先前在FPKc中发现的ES与FPKc具有相似的作用。因此我们可以得出结论，ES作为FPKc中含量最高的成分之一，可能也是活性成分之一。
结论/意义：FPKc可抑制SW - 480细胞的迁移，诱导SW - 480细胞G1期阻滞并引起ROS介导的凋亡效应。而ES可能是整个过程中的有效成分之一。
Zotero 插件