罗汉果醇抗肿瘤活性及其作用机制研究

Activity and mechanism of anticancer properties of mogrol

关键词:  罗汉果醇, 抗肿瘤, 细胞增殖, 细胞凋亡, 凋亡基因

Key words:  mogrol, antineoplastic, cell proliferation, apoptosis, apoptotic genes

摘要:

罗汉果醇是罗汉果皂苷的苷元,有研究报道罗汉果皂苷V具有防癌抑癌作用。该研究采用噻唑蓝实验(MTT法)检测罗汉果醇对不同肿瘤细胞增殖的抑制情况,以及不同浓度的罗汉果醇对CNE1细胞的增殖抑制率; 应用细胞克隆形成实验进一步验证罗汉果醇对CNE1细胞增殖的抑制作用; 采用Annexin V/PI 双染法检测罗汉果醇对CNE1细胞凋亡的影响; 以实时定量PCR技术检测罗汉果醇对CNE1细胞中Caspase-3、Survivin、 Bax和Bcl-2基因的mRNA 表达水平的影响。结果表明:罗汉果醇能显著抑制DU145、HepG2、A549、CNE1、CNE2细胞的增殖,其中对CNE1细胞增殖的抑制作用最为显著,并呈剂量依赖性,其半数抑制浓度IC₅₀为(81.48 ± 4.73)μmol·L^-1; 通过对CNE1细胞进一步的克隆形成实验,也验证了这一点; Annexin V/PI 双染法可见随着浓度的增加,凋亡比例增加; 实时定量PCR技术检测显示罗汉果醇处理后,促凋亡基因Caspase-3、Bax的表达增加,抗凋亡基因Survivin、Bcl-2的表达减少。因此,罗汉果醇可能是通过促进Caspase-3、Bax等促凋亡基因和抑制Survivin、Bcl-2等抗凋亡基因的表达,来诱导肿瘤细胞凋亡,进而发挥抗肿瘤活性。

Abstract:

Mogrol is the aglycone of mogrosides, and it is reported that Mogroside V has good cancer prevention and anti-cancer efficacy. This study was to explore the inhibitroy effects of mogrol against proliferation of human nasopharyngeal cancer CNE1 cells, and the molecular mechanism of apoptosis induced by mogrol. The inhibitory activities of different tumor cells were assessed by MTT assay. The concentration-dependent inhibition by mogrol on the most sensitive CNE1 cells were further studied. Cell clonogenicity was detected by colonyforming assay to further verification the inhibition of mogrol on the proliferation of CNE1 cells. And the apoptosis was examined by Annexin V/PI double staining. To further analyze the apoptosis mechanism of cell apoptosis, we detected the Caspase-3, Survivin, Bax and Bcl-2 mRNA expression levels in CNE1 cells after treating with mogrol by real time-PCR. The results showed that mogrol could significantly inhibit the proliferation of DU145, HepG2, A549, CNE1, CNE2 cells, the inhibitory effect of CNE1 cells was the most significant and in a dose-dependent manner, the IC₅₀ was(81.48 ± 4.73)μmol·L^-1. The colonyforming assay also verified that mogrol could inhibit the proliferation of CNE1 cells. Apoptosis of CNE1 cells was examined by Annexin V/PI double staining, the apoptosis rate was increased with concentration. Real time-PCR showed that mogrol could promote the expression of Caspase-3, Bax genes and inhibit the expression of survivin, Bcl-2 genes. Therefore, mogrol can probably induce the apoptosis of tumor cells, and result in anti-tumor activity, by promoting the expression of Caspase-3, Bax and other pro-apoptotic genes and inhibiting the expression of survivin Bcl-2 and other anti-apoptotic genes.