增加丹參癒合組織中丹參酮含量以及丹參酮 I 抑制非小細胞肺癌作用之研究

Abstract

Salvia miltiorrhiza Bunge, generally known as Danshen, the perennial herb is a traditional Chinese herbal medicine with multiple functions. However, its systematic study on pharmacogenomics has not been well established. In this study, by using an omnibus strategy, we investigate Agrobacteria transgenic, ingredients inducible, T-DNA activation tagging and microarray system, as well as the anti-cancer and anti-invasive mechanisms in non-small-cell lung cancer (NSCLC). We have set up a zeatin/2-4D inducible system of Danshen active ingredients, and confirmed by HPLC. Furthermore, the regeneration and Agrobacterium-transgenic systems were established by a BA regenerative and selective medium. The results of indicated that there are more than six independently transgenic cell lines with red color from 1435 transgenic lines while grown in non-inducible medium. ATM lines T1–T6 showed significant increases in the yields of tanshinone-I (up to 43-fold), tanshinone-IIA (up to 26-fold) and cryptotanshinone (up to 104-fold) compared with those of the nontransgenic lines on 2,4-D medium. Interestingly, the yield of cryptotanshinone from line T4 on 2,4-D medium was two times higher than that of the nontransgenic lines on trans-zeatin riboside medium. For the pharmacologic study of tanshinones, we found that the active ingredients including tanshinoneⅠand tanshinoneⅡA can significantly inhibit cell growth, migration, invasion, gelatinase activity, and IL-8 transcriptional regulation of human lung adenocarcinoma cell CL1-5 with highly invasive capability. Interestingly, only TanshinoneⅠ could significantly reduce AP-1 and NF-κB binding activity of CL1-5 cells measured by EMSA, as well as the tumorigenesis of mice. However, tanshinoneⅡA revealed a much stronger pro-apoptotic activity than TanshinoneⅠ measured by flow cytometry analysis. To further interpret the mechanisms of Danshen on anti-metastasis and anti-angiogenesis, human cDNA microarray was applied to this study. The results showed that tanshinoneⅠcan suppress several metastasis-related genes including PDGF-β, Shc, Shb, ephrin-A1, Rab8, MAPKK3, CD4 and Rac 1. Furthermore, tanshinone I significantly block increased the expression of focal adhesion proteins α-actinin and β-catenin, and upregulated phosphorylation of cofilin through the inhibition of Rac1 expression and activity. The in vivo antimetastatic effects of tanshinone I were evaluated in SCID mice following a tumor xenograft implantation of CL1-5 cells. Tanshinone I (0.3 mg per kg daily) inhibited lung metastasis and cofilin dephosphorylation in CL1-5-bearing mice. These data suggested that Danshen has the potential for increasing the yields of tanshinons. On the other hand, tanshinone I may have the potential for anti-cancer and anti-metastasis, and may be helpful in treating the cancer, as a medicine or an assistant chemotherapy in lung cancer.