摘要
探究桦褐孔菌液体深层发酵所得菌丝体中三萜类成分的抗肿瘤活性。采用高效液相色谱法鉴定桦褐孔菌液体深层发酵产物中含有5种已知具有肿瘤细胞生长抑制活性的三萜类成分,采用MTT比色法分析三萜类成分对卵巢癌细胞ES2、SKOV3和肺癌细胞A549和PC9的生长活性的影响,通过细胞周期和细胞凋亡实验研究三萜类成分对卵巢癌细胞和肺癌细胞增殖的影响。结果表明:经过三萜类成分处理3d和6d的肿瘤细胞生长活性均被明显抑制,对卵巢癌细胞ES2抑制的IC50分别为21.1μg/mL和17.6μg/mL,对SKOV3抑制的IC50值分别为33.8μg/mL和10.9μg/mL;对肺癌细胞PC9抑制的IC50分别为21.4μg/mL和19.7μg/mL、对A549抑制的IC50值分别为28.5μg/mL和23.5μg/mL。细胞周期实验结果表明三萜类成分对ES2、A549、PC9、SKOV3的阻滞可能发生在G0/G1期,在16.0μg/mL的三萜类成分处理下,PC9、A549、ES2、SKOV3处在G1期的细胞比例与对照组中各细胞G1期的占比分别提高32.3%、45.7%、10.9%、6.2%,S期的细胞数降低19.1%、57.3%、23.1%、17.5%。细胞凋亡结果表明,经过药物处理2d后,三萜类成分能够有效的诱导卵巢癌细胞ES2、SKOV3和肺癌细胞A549和PC9的细胞凋亡,进一步验证桦褐孔菌三萜类成分具有肿瘤抑制活性。
This study aims to investigate the anticancer activity of triterpene from submerged fermentation of Inonotus obliquus.HPLC experiments showed that submerged fermentation products of Inonotus obliquus contained five triterpenes which could inhibit tumor cell growth.MTT colorimetric assay was used to analyze the influence of triterpenes on growth activity of ovarian cancer cells ES2 and SKOV3, and lung cancer cells A549 and PC9.Cell cycle and cell apoptosis experiments were carried-out to investigate the effect of triterpenes on proliferation of ovarian cancer cells and lung cancer cells.The results showed that,growth activity of the cancer cells treated by triterpenes for 3d and 6d was obviously inhibited.The IC50 values of ES2 were 21.1μg/mL and 17.6μg/mL,respectively;the IC50 values of SKOV3 were 33.8μg/mL and 10.9μg/mL,respectively;the IC50 values of PC9 were 21.4μg/mL and 19.7 μg/mL,respectively;the IC50 values of A549 were 28.5μg/mL and 23.5μg/mL,respectively.The results of the cell cycle experiments showed that the inhibition of ES2,SKOV3,PC9 and A549 may occur in G0/ G1 phase.After treatment with 16.0μg/mL triterpenes,the proportion of cells in G1 phase of PC9,A549,SKOV3 and ES2 increased by 32.3%,45.7%,10.9%and 6.2% respectively,compared with the control group,and the proportion of cells in S phase reduced 19.1%,57.3%,23.1% and 17.5%,respectively. Apoptosis results showed that after treatment with triterpenoid for 2d,triterpenes could effectively induce apoptosis of ES2,SKOV3,PC9 and A549,which further verifies the triterpenoids of Inonotus obliquus possess tumor cell inhibition activity.
作者
金黎达
欧文斌
徐向群
JIN Lida;OU Wenbin;XU Xiangqun(School of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018)
出处
《浙江理工大学学报（自然科学版）》
2019年第1期98-105,共8页
Journal of Zhejiang Sci-Tech University(Natural Sciences)
基金
国家自然科学基金项目(81728012)
浙江省自然科学基金项目(LY16B020013,LY18H160065)
关键词
桦褐孔菌
液体深层发酵
三萜
肿瘤细胞抑制
Inonotus obliquus
submerged fermentation
triterpene
tumor cell inhibition
