Doenjang is a major fermented soy-based food in Korea. Recent investigations have shown that fermented soybean foods have immunity-enhancing, anti-cancer, anti-obesity and anti-diabetic effects. Several studies also have reported that genistein and daidzein, which are easily absorbed in the body are produced in larger quantities in aged doenjang. The purpose of this study was to evaluate the variations in the anti-cancer effects of commercialized doenjang as it ages. Four groups were formed for this study according to aging periods of doenjang, namely short (under 5 years, S group), mid (under 10 years, M group), long (under 15 years, L group) and very long (over 15 years, E group). The anti-cancer effects of doenjang were determined by cell cytotoxicity assays in A549, YAC-1, and HepG2 cancer cell lines. Also, NK cell activity and splenocyte proliferation were assayed for cancer immunotherapy. The quantities of phenolic compounds in doenjang at different ages were also measured. The results showed that the anti-cancer effects increased in the S and M groups for all three cancer cell lines. Interestingly, similar to this result, splenocyte proliferation and NK activity were also the highest in the S and M groups. In contrast, the E group showed significantly reduced splenocyte proliferation. The quantity of phenolic compounds was similar to that of the anti-cancer results. Collectively, these results suggest that the fermentation period of doenjang plays a very important role in determining its anti-cancer effects.
In this paper, we investigate to determine quality characteristics, fatty acid composition and cytotoxic effect of extracts and fractions from whole Lycopus lucidus Turcz. roots. Additionally, we evaluated cytotoxic activity against the growth of human fibrosarcoma cells (HT-1080) and human gastric adenocarcinoma (AGS), human colon cancer cell (HT-29) lines using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Acetone+methylene chloride (A+M) and methanol (MeOH) extracts from L. lucidus Turcz. were obtained through solvent extraction. Then we further fractionated both extracts with n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water. In fatty acid composition, L. lucidus Turcz. contained 33.2% of 18:1n-9 and 1.81% of 18:3n-3, respectively. The incorporation of treatment with A+M and MeOH extracts and n-hexane, 85% aq. MeOH, n-butanol (n-BuOH) and water fractions dose-dependently increased cytotoxicity against the growth of HT-1080 and AGS, HT-29 cancer cells (p<0.05). The A+M extract had a higher inhibitory effect on the growth of all cancer cells in comparison to MeOH extract. Among the fractions, the 85% aq. MeOH and n-hexane fractions showed a higher inhibitory effect after proliferating the three cancer cells. These results suggest that the 85% aq. MeOH and n-hexane fractions have a potential to inhibit the growth of human cancer cell lines.
Ganoderma lucidum has been traditionally used as a medicine for treatment of bronchitis, arthritis, and high blood pressure, and it has been reported to display many biological activities including anticancer and immune activities. Since mushroom mycelium is known to have excellent biological activities together with mushroom fruiting body, studies on biological activities of mushroom mycelium have been actively conducted. Thus, the present study compared the biological activities before and after the cultivation of Ganoderma lucidum mycelium on Atractylodes rhizoma. When the radical scavenging activity was assessed by the DPPH assay, ARGL (ethanol extract of Atractylodes rhizoma mycelium fermented with Ganoderma lucidum) showed radical scavenging activity of 5.58~82.56% at concentrations of 10~500 μg/assay, while AR (ethanol extract of Atractylodes rhizoma) showed radical scavenging activity of 5.27~72.08% at the same concentrations. When measured by using the ABTS assay, ARGL showed higher radical scavenging activity than AR, which was consistent with the result obtained by the DPPH assay. In the MTT assay, the cytotoxicity of ARGL against all cell lines was higher than that of AR. In particular, the cytotoxicities of AR and ARGL against Hep3B at a concentration of 400 μg/assay were 71.81% and 86.40%, respectively. In addition, the result obtained by the SRB assay was consistent with the result obtained by the MTT assay. According to the results mentioned above, there is a high probability that medicinal herb cultures using mycelium can be used as sources of functional foods since the cytotoxicities against cancer cells and antioxidant activities increased when the mycelium was fermented with Atractylodes rhizoma.
Lycorine, a natural alkaloid extracted from the Amaryllidaceae plant family, was reported to various physiological and pharmacological effects including anti-cancer activity. Nevertheless, there is no report of the anticancer effect of lycorine in oral cancer cells. The effects of lycorine on cell proliferation and apoptosis were examined through trypan blue exclusion assay, 4’-6-diamidino-2-phenylindole (DAPI) stain, Live/Dead assay, Western blot analysis and RT-PCR. Lycorine suppressed cell viability and induced apoptosis in MC3 and HSC-3 cell lines. Lycorine decreased survivin protein but did not affect its mRNA. It regulated survivin through accelerating protein degradation in a time-dependent manner although neither proteasome nor lysosome was not associated with lycorine-mediated protein degradation. Collectively, our results suggest that lycorine may be a potential therapeutic anti-cancer drug candidate for the treatment of human oral cancer.
실험에서는 꽃송이버섯 (Sparassis crispa, formerly S. crispa) 에탄올 추출물의 소수성 분획을 분리하고 각 분획의 DPPH 항산화 활성과 위암 (AGS), 폐암 (A529), 간암 (HepG2) 세포주를 이용한 항암 활성을 MTT assay 를 통해 확인 하였다. SOCC를 사용하여 총 18개의 fraction으로 분획하였고 TLC와 세포주를 이용한 항암 활 성 확인을 통하여 5개의 fraction으로 압축하였다. 항암 활성이 높은 5개의 fraction은 HPLC-MS를 통해 각 분획 물을 분석한 결과 항산화 활성을 보이지 않았으며 약 181.0의 분자량을 가진 물질이 지표물질로 확인되었으므 로 이 물질의 화학식 동정을 위하여 추가실험이 필요하다. 세포주를 이용한 항암실험 결과 꽃송이버섯 추출물은 위 암 (AGS), 폐암 (A529), 간암 (HepG2) 세포주 모두에서 양성대조군인 paclitaxel보다 낮은 세포 생존율을 보여 주 었으며(IC50 value), 이것은 추후 꽃송이버섯 추출물에 포 함된 항암 물질 분리 연구를 위한 기초연구 결과로서 무 척 의미가 크다고 할 수 있다.
Lung cancer caused by diverse changes in cells resulted by exposure to carcinogens found in tobacco smoke, the environment, or sequential accumulation of genetic changes to the normal epithelial cells of the lung. An assessment was made of the anti-proliferative activity of constituents from silkworm feces against 11 human cancer cell lines, including A549 and H727 lung cancer cell lines, using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The ethanol extract of silkworm feces was proved to have anti-proliferative activity against all 11 species of human cancer cell lines. The biologically active constituent was characterized as vomifoliol (blumenol A) (1) and stigmasterol (2) by spectroscopic analysis ,including MS and NMR. In conclusion, global efforts to reduce the level of antitcancer agents justify further studies on the silkworm feces-derived materials containing vomifoliol and stigmasterol as potential anticancer products or lead compounds for the prevention or eradication from human lung cancer.
The anti-proliferative efficacy of t,t-conjugated linoleic acid (t,t-CLA), c9,t11-CLA, and t10,c12-CLA was compared in several human cancer cell lines. Gastric NCI-N87, liver Hep3B, pancreas Capan-2, and lung NCI-H522 cancer cells were incubated with 50 μM CLA isomers over a period of 6 days. The t,t-CLA inhibited the growth of all cancer cell lines to different extents, but c9,t11-CLA and t10,c12-CLA inhibited or stimulated the growth of the cancer cell lines. NCI-N87 cells were the most sensitive to growth inhibition and apoptosis from all CLA isomers tested. In NCI-N87 cells, CLA isomers reduced the release of arachidonic acid (AA) via the inhibition of cytosolic phospholipase A2 (cPLA2 ) activity, consequently reducing the production of PGE2 through the inhibition of cyclooxygenase-2 (COX-2). The efficacies of CLA isomers were in the following order (from most to least effective): t,t-CLA, t10,c12-CLA and c9,t11-CLA. Overall, these results imply that the anti-proliferative efficacy of t,t-CLA on cancer cells, especially NCI-N87 cells, was greater than other CLA isomers due to its induction of apoptosis through the inhibition of cPLA2 and COX-2 activities.
For investigate intracellular function and role of genes in the biological processes, various gene delivery methods into cell have been developed. Many studies performed to construct optimum conditions of gene delivery into cells and tissues. In this study, we examined efficiency of gene delivery-complexed with cationic lipid vector in human cancer cell lines. GFP plasmids were complexed with cationic lipid and transfected into human cancer cell lines at different concentrations. And then, expression of GFP was analysed with fluorescent microscope and FACS. To determine efficiency of gene delivery, we investigated GFP expression level in various cancer cell lines. GFP expression cells were not shown in hepatocellular carcinoma cell line HepG2 and lung carcimona cell line A549 after 24hr transfection, while, GFP expression cells were observed at 500ng concentration after 48hr transfection. In colorectal carcinoma cell line HCT116, GFP expression cells were observed at 100ng and 500ng concentrations after 24hr transfection and slightly increased at 48hr. After transfection into ovary adenocarcinoma cell line SKOV3, we could found that many cells expressed GFP at 500ng concentration after 24hr and highly elevated GFP expression cells after 48hr. For further evaluate gene expression level, we confirmed GFP expression level by using FACS analysis after 48hr transfection. As a result, HepG2 was expressed GFP in very low level at 10ng, 100ng, and 500ng concentrations. We also identified that GFP was expressed low level at 10ng and 100ng in HCT116 and A549, but highly increased at 500ng concentration to 14.19% and 16.57%, respectively. In case of SKOV3, GFP expression was highly elevated to 13.14% at 100ng and 58.10% at 500ng compared with 10ng transfection. By Comparing efficiency of gene expression among cancer cell lines, GFP expression was similar with cell lines at 10ng transfection, but significantly differed from cell lines at 500ng higher concentration. Additionally, GFP expression level of SKOV3 was showed about 10 fold higher than HepG2, and about 4 fold higher than HCT116 and A549 at 500ng. These results demonstrated that efficiency of gene delivery-complexed with cationic lipid vector was the highest in SKOV3, while HepG2 was showed the lowest efficiency. Taken together, we could determined that efficiency of gene delivery into cells differed from each human cancer cell lines. Our study suggest that cellular properties should be considered in gene delivery-complexed with cationic lipid vector to improve cellular expression efficiency of gene.
Many methods have been developed for more efficient gene delivery and expression in human cells. A number of studies have been performed in achieving successful gene delivery and expression conditions. We investigated differential gene expression patterns after delivery adenoviral vector containing green fluorescent protein(GFP) gene into human cancer cell lines. We constructed recombinant adenoviral Ad-CMV-GFP containing CMV promoter and GFP gene. The efficiency of gene expression was assessed by observation GFP expressing cells using fluorescent microscopy after transfer of Ad-CMV-GFP in concentrations of 0.1μl. 1μl. 10μl. At first, we evaluated expression patterns of gene in several human cancer cell lines, gastric adenocarcinoma cell line AGS was showed high level of GFP expression compared with colorectal adenocarcinoma cell line HT-29. After transfer 0.1μl of Ad-CMV-GFP in AGS, we could found that GFP expression cells were observed in next day and highly increased 2 days. While, small number of GFP expressing cells were examined in HT-29 and SNU-C4. Therefore, these data showed that AGS was expressed the highest level of GFP and almost AGS cells seems to express GFP in concentration of 1μl of Ad-CMV-GFP. GFP expression pattern in HT-29 reveal that expression was low in next day after gene transfer but significantly increase expression level in 2 days. In case of SNU-C4, GFP expression increased with increasing concentration of Ad-CMV-GFP and t ransfer times. For examine effects of transfer times in small amount gene, we transfer in concentration of 0.1μl Ad-CMV-GFP and detected GFP expression patterns after 2 days or 4 days. As a result, expression level of GFP in AGS was increase about 2 fold after 4 days compared with 2 days, but any difference of GFP expression levels were not showed in HT-29 and SNU-C4. Our study suggested that adenovirus was very efficient gene transfer vector for gene expression in human cancer cell lines. In addition to, we also demonstrated that gene expression patterns was dependent on each human cell lines. Therefore, further studies will be needed to confirm the optimum conditions for efficient gene delivery and expression in each target cell lines with consideration to cellular properties.
The aim of this study was to investigate the cytotoxic effect and its mechanism on Radix Aconiti(RA) extract in lung cancer cell lines. RA extract treatment decreased the cell viability in a dose-dependent fashions in lung cancer cells including A549, H460, H23 and H157 cells. Many investigators reported that A549 and H460 cells expressed wild-type p53, but H23 and H157 cells preserved mutated p53. After treatment with RA extract in A549 and H460 cells, we measured the expression of p53 protein levels using Western blot. analysis. In both cells treated with RA extracts, p53 protein expressions were increased in a dose-dependent manner. In our experiments, RA extracts also have cytotoxic effects in H23 and H157, which have mutated p53. Treatment with RA extract decreased bcl-2 protein expressions in both cells. These results suggest that RA extracts have cytotoxic effects via p53 expression increase and bcl-2 inhibitable pathways in A549, H460 cells and H23, H157 cells, respectively.
Alterations in cell surface receptors and adhesion molecules which regulate cell-cell and cell-matrix interactions have been 뻐plicated in tumor processes. In order to investigate the effect of integrin expression on the invasiveness of oral sqllamous cell carcinoma, integ띠1 expression in the celllines such as SCC-4, SCC-9, SCC-15, and SCC-25 was analyzed, and the comparison between cell adhesion assay towards extracellular matrix proteins and in vitro invasion assay following inhibition of the functional domain of the integrins using blocking antibodies against the specific integrins 낀 nd Arg-Gly-Asp (RGD) peptide were carried out. The expression of integrin a 2, a 3, a 6 was detected in all oral squamous cell carcinoma celllines. In contrast, the expression of a vß6 integrin is detected in SCC-4 and SCC-9, not in SCC-1 5 and SCC-25. 까1e adhesion of SCC-4 cell line to collagen 1, laminin, and fibronectin was significantly reducecl by σeatment with a 3-, a 6-, and a vß6-blocking antibody, respectively (p (0.05). 꺼.1e invasion of SCC-4 cell line throllgh Matrigel was significantly reduced by treatment with v 6-blocking antibody and RGD pepticle (p(0.05). These results sllggested that specifìc integrins play an in1portant role in the process of adhesion and invasion of oral squamous cell carcinoma cells and the expression of a vß6 integrin is believed to the enhance its invasivene잃.
chemosensitivity test of Geungsonojukwhan-Bijukbang was performed on the three different human cancer cell lines originated from liver, cervix and colon tissue, namely Hep 3B, Hela and HCT-15, which have similar doubling times. Semiautomated sulforhodamine B(SRB) assay appears to offer an valuable tool for chemosensitivity of unknown compounds, since it is a simple, valid and inexpensive method of assessing drug monitoring for large samples in a short time. The results obtained in this study were as follows 1. Good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. As a result of exposure to Geungsonojukwhan, the proliferation of Hela cell and Hep 3B cell was slightly decreased in Geungsonojukwhan-Bijukbang(GIP), Geungsonojukwhan-Pejukbang(LUP) and Geungsonojukwhan-Sinjukbang(RTP). 3. As a result of exposure to Geungsonojukwhan, GIP showed better anticancer effect to HCT-15 cell lines than those of LUP and RTP. 4. The extract of Geungsonojukwhan-Bijukbang in 40℃ were more effective in cytotoxic response than those in 100℃. 5. The research showed that the higher concentration the more effective in the inhibition of proliferation of the cancer cell lines, however, the cytotoxic effect of Geungsonojukwhan-Bijukbang in the concentration of 1.60㎎/㎖ and 3.20㎎/㎖ showed the most effective inhibition rate according to the increase of concentration.