Xu Tao, Jin Fa, Wu Keren, Ye Zhipeng, Li Ning
Department of Hepatobiliary Surgery, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou, Zhejiang 310005, P.R. China.
Exp Ther Med. 2017 Nov;14(5):5129-5134. doi: 10.3892/etm.2017.5168. Epub 2017 Sep 21.
The aim of the present study was to determine the ability of oridonin to enhance the anticancer activity of lentinan (LNT) in SMMC-7721 human hepatoma cells by using various techniques, including MTT, flow cytometry, quantitative PCR and western blot assays. The results demonstrated that 20 µg/ml was a non-toxic concentration of oridonin for L02 normal liver cells and SMMC-7721 cells, while 0-200 µg/ml of LNT only had anti-proliferative effects on SMMC-7721 cells. LNT at 100 and 200 µg/ml inhibited the growth of SMMC-7721 cells by 22.8 and 60.0%, respectively, and after addition of 20 µg/ml oridonin, the inhibitory rate of 100 and 200 µg/ml LNT was increased to 47.2 and 80.7%, respectively. Oridonin (20 µg/ml) + LNT (200 µg/ml)-treated SMMC-7721 cells showed the highest apoptotic rate, which was 40.5±2.5%, which was higher than that of cells treated with LNT only. LNT raised the mRNA and protein expression of caspase-3, -8 and -9 as well as B-cell lymphoma 2 (Bcl-2)-associated X protein, p53 and p21, while reducing the expression of Bcl-2, Bcl extra large protein, epidermal growth factor (EGF) and EGF receptor expression in SMMC-7721 cells as compared to that in control cells. Treatment with 20 µg/ml oridonin and 200 µg/ml LNT increased these changes of gene expression. From the obtained results, it may be concluded that oridonin raised the anti-cancer effects of LNT in SMMC-7721 cells. Oridonin may also be used as a sensitizing agent to increase the anticancer activity of LNT .
本研究的目的是通过使用多种技术,包括MTT、流式细胞术、定量PCR和蛋白质印迹分析,来确定冬凌草甲素增强香菇多糖(LNT)对SMMC-7721人肝癌细胞抗癌活性的能力。结果表明,20μg/ml的冬凌草甲素对L02正常肝细胞和SMMC-7721细胞无毒,而0-200μg/ml的LNT仅对SMMC-7721细胞有抗增殖作用。100和200μg/ml的LNT分别抑制SMMC-7721细胞生长22.8%和60.0%,加入20μg/ml冬凌草甲素后,100和200μg/ml LNT的抑制率分别提高到47.2%和80.7%。冬凌草甲素(20μg/ml)+LNT(200μg/ml)处理的SMMC-7721细胞凋亡率最高,为40.5±2.5%,高于仅用LNT处理的细胞。与对照细胞相比,LNT提高了SMMC-7721细胞中半胱天冬酶-3、-8和-9以及B细胞淋巴瘤2(Bcl-2)相关X蛋白、p53和p21的mRNA和蛋白表达,同时降低了Bcl-2、Bcl-2超大蛋白、表皮生长因子(EGF)和EGF受体的表达。用20μg/ml冬凌草甲素和200μg/ml LNT处理增加了这些基因表达的变化。从所得结果可以得出结论,冬凌草甲素提高了LNT对SMMC-7721细胞的抗癌作用。冬凌草甲素也可作为增敏剂来增强LNT的抗癌活性。