灵芝酸单体调控乳腺癌细胞生长及相关基因表达的研究

Study of Ganodema Acid Monomer Regulating the Growth of Breast Cancer Cells and Related Gene Expression

目的:探讨灵芝酸单体A,B调控乳腺癌细胞生长以及相关基因PI3K,AKT,mTOR,PTEN的表达。方法:采用CCK8试剂盒分别检测对照组和不同浓度灵芝酸A,B(0、1、5、10、20、50、100μmol·L^(-1))对乳腺癌MDA-MB-231和乳腺癌MCF-7细胞的增殖影响。每组设3个复孔,分别接种于96孔板中,分别于培养24 h和48 h检测其OD值。结果:CCK8法检测,与对照组相比,随着灵芝酸浓度增加,细胞存活率下降,当灵芝酸浓度为100μmol·L^(-1)时,细胞存活率最低(p<0.05或p<0.01),细胞抑制作用最强。同时随着药物处理时间的增加,对细胞的抑制作用更加明显。表明灵芝酸调控乳腺癌细胞增殖作用与浓度和时间有紧密相关性。灵芝酸A和灵芝酸B处理MCF-7和MDA-MB-231两种细胞24 h、48 h,提取细胞RNA,通过反转录获得c DNA,再经Real-timePCR检测相关基因表达。结果表明灵芝酸单体A,B可以下调两种乳腺癌细胞pi3k、akt、mtor mRNA的表达,上调pten的表达。结论:灵芝酸可以使乳腺癌细胞增殖活性降低,可能通过调控PI3K/AKT/mTOR通路的活性,从而抑制肿瘤细胞的生长。

Objective: To investigate the regulation of Ganoderma acid monomer A and B on the growth of breast cancer cells and the expression of PI3 K, Akt,mTOR and PTEN. Methods: CCK8 kit was used to detect the proliferation of MDA-MB-231 and MCF-7 cells in breast cancer. Three double holes were set up in each group, and then inoculated in 96 hole plate, and their odvalue was detected 24 hours and 48 hours respectively. Results: compared with the control group, the cell survival rate decreased with the increase of Ganoderma acid concentration. When the concentration of Ganoderma acid was 100 μmol·L^(-1), the cell survival rate was the lowest(P<0.05 or P<0.01), and the inhibition effect was the strongest. At the same time, with the increase of the treatment time, the inhibition of the cells is more obvious. It was found that Ganoderma acid had a close relationship with the concentration and time of proliferation of breast cancer cells. The RNA was extracted from MCF-7 and MDA-MB-231 cells for 24 hours and 48 h after treatment with Ganoderma acid A and Ganoderma acid B, and then the related gene expression was detected by real-timepcr. The results showed that Ganoderma acid monomer A and B could down regulate the expression of PI3 K, Akt and mTOR mRNA and up regulate the expression of PTEN. Conclusion: Ganoderma acid can reduce the proliferation of breast cancer cells, and may inhibit the growth of tumor cells by regulating the activity of pi3 k/akt/mtor pathway.