Suicide gene transfer has been study extensively for therapies in various human diseases. We can evaluate cellular activity of thymidine kinase and cytotoxic effect in colon cancer cells after suicide gene transfer. We observed cellular expression of green fluorescence protein after transfer with adenovirus into colon adenocarcinoma HCT-15 cells. After transfer HSVtk, we also estimated thymidine kinase activity using [3H]-penciclovir and cellular cytotoxicity by MTT assay. After transfer green fluorescence protein into HCT-15 cells, we could observed fluorescence expression in 10 moi concentration. Expression level of green fluorescence protein markedly increased in 30 moi and most of HCT-15 cells expressed green fluorescence protein in 100 moi. By infection with HSVtk in HCT-15 cells and HT-29 cells, thymidine kinase activity in HCT-15 cells was about two fold higher than that HT-29 cells. Thymidine kinase activity at 1 moi concentration makes no difference with 0 moi in both cells. At 10 moi concentration, thymidine kinase activity increased about three fold compared with 1moi in HCT-15 cells, but not observed high increase in HT-29 cells. Thymidine kinase activity at 100 moi showed about three fold increase in HCT-15 cells and one and a half fold in HT-29 cells compared with 10 moi. By treatment of HSVtk at various mois and ganciclovir to HCT-15 cells, we could find that increased cytotoxic effect according to HSVtk concentration. Cellular cytotoxic effect was slightly appeared at 5 moi concentration and intensively increased at 30 moi concentration, dead colon cancer cells were reached about 30% of total colon cancer cells. Cellular cytotoxic effect was consistently increased until 50 moi, and about 50% of cells at 100 moi and less then 50% of HCT-15 cells at 200 moi were survived. Finally, we can identify that suicide gene transfer into HCT-15 cells is performed according to concentration of suicide gene and thymidine kinase activity also increase with HSVtk concentration in both HCT-15 cells and HT-29 cells. Additionally, we also find that suicide gene therapy by HSVtk with ganciclovir intensively increase cellular cytotoxicity in colon cancer cells. Therefore, our findings suggest that suicide gene therapy by HSVtk can affect cytotoxicy for colon cancer cells and eventually seems to influence therapeutic efficacy.

팽이버섯 11계통을 사용하여 polyphenol 및 β-glucan 함량을 분석하고, 생리활성으로 항산화 및 항암, 항고혈압, 항당뇨, 항염활성을 측정하였다. Polyphenol 함량에서는 CBMFV-65가 244.74 mg%로 가장 많은 함량을 나타내었고 전 품종에서 전반적으로 100 mg% 이상의 함량을 나타냈다. β-glucan 함량에서는 CBMFV-41에서 27.37%로 가장 높았으며, 그 다음으로 CBMFV-30 27.21%, CBMFV-65 27.11%의 순서로 높은 β-glucan 함량을 나타냈다. 전자공여능에서는 CBMFV-66이 91.74%로 가장 높은 DPPH 소거활성을 나타냈으며 전체적으로 70%의 높은 소거활성을 보였다. 세포독성 실험에서는 폐암세포에 대한 세포 저해활성이 가장 컸으며, 폐암세포와 대장암세포 모두 CBMFV-30에서 각각 76.07%, 67.05%로 가장 높은 세포 저해활성을 나타냈다. ACE 저해활성에서는 CBMFV-65에서 10.96%를 나타냈고 나머지 품종은 10%미만의 저해활성을 나타냈다. 항당뇨 활성에서는 CBMFV-41에서 63.57%의 효소 저해율이 측정되었고, CBMFV-63은 10.98%로 가장 낮은 효소 저해활성을 나타냈다. 마지막으로 항염활성에서는 CBMFV-41에서 61.44%의 nitric oxide 억제율이 측정되었다.

맛버섯 10계통의 에탄올 추출물에 대한 polyphenol 및 β-glucan 함량을 분석하고, 생리활성으로 항산화 및 항암, 항고혈압, 항당뇨, 항염활성을 측정하였다. Polyphenol 함량에서는 전 계통에서 전반적으로 40 mg% 함량 이상이였고, 맛버섯 M-1548이 61.50±0.59 mg%로 가장 높았다. β-glucan 함량에서도 맛버섯 M-1548에서 37.2±1.12%로 가장 높았으며, 그 외에 맛버섯 M-900에서 36.35±1.11%, M-1630에서 36.24±1.27%의 순서로 높은 β-glucan 함량을 나타냈다. 항당뇨 활성에서도 역시 맛버섯 M-1548이 13.78±0.56%로 가장 높은 효소 저해율을 보였으나 항염 활성에서는 맛버섯 M-1630이 56.59±7.11%로 가장 높은 nitric oxide 저해율을 보였으며 맛버섯 M-1548은 26.21±0.5%로 미미한 저해율을 보였다. 전자공여능 및 ACE 저해활성, nitrite 저해활성은 효과가 없거나 미미한 활성만을 나타냈다. 세포독성 실험에서는 1 mg/mL로 처리시 폐암세포에 대해 전반적으로 30%이상의 세포 사멸율을 보였으며, 자궁경부암세포에서도 맛버섯 M-1548에서 10.05±0.44%의 세포 사멸율을 보였다. 그러나 대장암세포와 정상세포인 섬유아세포에서는 세포 사멸율이 나타나지 않았다. 따라서 맛버섯 10계통은 폐암세포와 자궁경부암세포에 세포 독성을 나타내는 것으로 보아 암세포에 대한 선택성을 갖고 있음을 알 수 있고 정상세포에 대해선 세포 독성을 나타내지 않는 걸 확인하였다.

Dendropanax morbifera Léveille (Araliaceae) is an endemic species growing in the south-western part of South Korea and has been used in folk medicine and health functional food. Several studies have indicated that extract of D. morbifera (DP) has cytotoxic activities on a number of human cancers, such as, breast cancer, lung cancer, hepatoma, and chorioepithelioma. Recently, polyacetylene and triterpene compounds have been isolated from the DP and showed to have anti-complement activity. β-Amyrin, α-amyrin, dendropanoxide, and β-sitosterol are isolated from DP. However, its biological activities in cancer have not yet been clearly elucidated. In this study, we evaluated the anti-cancer activity of isolated triterpenoids from the DP leaves by measuring the levels of cytotoxicity against MCF-7 human breast cancer and A549 human lung cancer cells. Six triterpenoids were isolated from the n-hexane fraction of DP leaves along with the known compounds. β-amyrin (1), α-amyrin (2), olean-12-en-3,24 β-diol (3), dendropanoxide (4), β-sitosterol (5), and stigmasterol (6). Compound 3 and 6 were isolated from DP for the first time. Cytotoxic activities of six compounds were evaluated against two human cancer cell lines by using the MTT in vitro assay. Among them, five compounds (1, 2, 4, 5, and 6) showed moderate cytotoxic activities toward the tested cell lines, and were safe to normal cells. Western blot analysis showed a decreased expression of anti-apoptotic protein Bcl-2 and increased levels of pro-apoptotic protein Bax in MCF-7 and A549 cells treated by β-amyrin and α-amyrin. Flow cytometry analysis confirmed that five compounds (1, 2, 4, 5, and 6) treatment increased populations of sub-G1 (apoptosis) phase. The results of the present study revealed that triterpenoids from DP have the potential for further development as anticancer agents.