| Summary: | 碩士 === 台南應用科技大學 === 生活應用科學研究所 === 100 === (I)
Colorectal cancer is the leading cause of cancer mortality, and metastasis is responsible for approximately 40% of death in colon cancer patients. Matrix metalloproteinases (MMPs) are key enzymes in the degradation of extracellular matrix, and MMP-2 and -9 are critical for cell migration leading to invasion and metastasis of cancer. The inhibition of MMP-2 and -9 is therefore considered that might depress the occurrence of tumor invasion and metastasis. Rosmarinic acid (RA) is a bioactive polyphenol that widely presented in Rosmarinus officinalis L. However, the literature regarding the effect of RA on invasion of cancer cells is still limited. In this study, we investigated the anti-invasion activity of RA against human colorectal adenocarcinoma Colo205 and HT-29 cells. The cells were treated with 0, 10, 50, 100, or 200 M of RA at 37C for 24 h, and the MMP-2, -9, and uPA activities as well as cell-matrix adhesion, motility, and invasion activities were determined. The MMP-2, -9, TIMP-1 and -2 mRNA levels were assayed by RT-PCR. The molecular signaling was determined by Weastern blot. The results showed that the MMP-2 and uPA activities of Colo205 and HT-29 cells were significantly inhibited by RA at a concentration of > 50 μM. The migration and invasion activities of Colo205 and HT-29 cells were also suppressed after treating with RA at a concentration higher than 50 M. The mRNA level of MMP-2 in HT-29 and Colo205 were significantly inhibited by RA at a concentration of > 50 M. The mRNA levels of TIMP-1 and -2 in HT-29 and TIMP-1 in Colo205 were significantly increased by RA. The signaling of p-ERK and p-p38 in Colo205 and HT-29 were signigicantly inhibited by RA. Furthermore, the activations of NF-κB and AP-1 in colorectal cancer cells were also suppressed by RA. Our results suggest that RA might be a bioactive with potential anti-invasive activity against colorectal cancer cells by inhibiting uPA and MMP-2 activity and reducing motility capabilities through inhibiting the activations of MAPKs signaling pathways and NF-κB and AP-1 transcription factors.
(II)
Tuberculosis (TB) is a contagious disease which causes a serious public health risk. WHO estimates that there were approximately 9,400,000 new TB cases globally in 2008, and South-East Asia Region accounted for 34% of incident cases. Multidrug-resistant TB (MDR-TB) is a particularly dangerous form of TB, which is responsible for the majority of failures in TB therapy. Mycobacterium tuberculosis is the major pathogeny of TB, and uridine diphosphate-glucose pyrophosphorylase (UGPase; EC 2.7.7.9) is the major enzyme that involved to the synthesis of cell wall. The inhibition of UGPase might depress the proliferation of the bacilli. Rosmarinic acid (RA), a polyphenol which is widely presented in natural plants of Lamiaceae and Boraginaceae family, has been reported that possesses several biological activities such as anti-virus, anti-inflammation, and anti-bacteria. The aim of this study was to investigate the inhibitory effects of RA on the viability of multidrug-resistant M. tuberculosis, and their impact on UGPase was also evaluated. Because the biological activity of a bioactive sometimes having an association with their antioxidant power, we therefore first determined the antioxidant activity of RA by trolox equivalent antioxidant capacity (TEAC) method. The viability of the bacilli was measured by counting the colony growing on 7H11 agar plates, and the expression of UGPase was performed by RT-PCR. The results showed RA having a strong antioxidant activity, and the viability of M. tuberculosis was reduced by treating with RA at a concentration of > 250 g/ml for 24 h. Furthermore, the expression of UGPase was also decreased by treating with 250 g/ml RA for 24 h. Based on the data mentioned above, it is suggested that RA can be used to inhibit the proliferation of multidrug-resistant M. tuberculosis, and the anti-proliferous activity of RA might be through inhibiting the expression of UGPase.
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