6-Gingerol exerts anti-tumor effects in various cancer cell models. We evaluated the effect of 6-gingerol on the growth of MCF-7 breast cancer cells and MCF-10A breast epithelial cells to determine whether any growth-inhibitory effects found were attributable to apoptosis, and to elucidate the underlying mechanism of action. 6-Gingerol inhibited the viability of both cell lines in a dose- and time-dependent manner; however, the degree of inhibition was greater in MCF-7 than MCF-10A cells. By flow cytometry, induction of dose- and time-dependent apoptosis was found, and the magnitude of apoptosis was also markedly greater in MCF-7 than MCF-10A cells. Expression of caspase-3 and poly (ADP-ribose) polymerase (PARP) was observed in MCF-7 cells treated with 6-gingerol, and further cleavage of PARP occurred in these cells. We suggest that 6-gingerol induces apoptosis in human breast cancer cells mainly by promoting caspase-3 expression and subsequent degradation of PARP.

Mushroom have long been valued as highly nutritious and tasty foods in many societies throughout the world. Itis known for biological activities including anti-inflammatory and anti-oxidative potential. However little is known about anti-cancer property. In this study, we investigated the anti-prostate cancer activity of mushrooms. For that, eight kinds ofmushrooms such as, T. matsutake, S. crispa, G. lucidum-US, G. lucidum-AS, C. cardinalis-BR, G. frondosa, P. linteus, U.esculenta were extracted with hot water. Among them, three kinds of mushrooms including T. matsutake, G. lucidum-US andC. militaris-BR extracts inhibited prostate specific antigen (PSA) expression in prostate cancer cell, LNCaP. These resultsdemonstrate that some of mushrooms inhibited PSA expression suggesting that the mushrooms might be a candidate for thetreatment of prostate cancer.

본 연구는 인체 유래 유방암세포인 MDA-MB-231 세포 증식을 억제하고 세포사멸을 유도하는 천연소재 발굴을 목적으로, 북한산 두릅 추출물을 MDA-MB-231 세포에 처리하여 세포사멸 및 작용기전을 규명하였다. 실험 결과, 두릅 추출물 처리 농도가 증가할수록 세포증식이 감소하였고, 세포질의 응축과 핵이 분절되는 등 apoptotic bodies를 형성하였다. 또한 유세포 분석기를 사용하여 MDA-MB-231의 세포주기가 억제되었고, 세포사멸의 특징인 sub-G1 수치가 증가되는 것이 관찰되었고, 두릅 추출물 농도가 증가함에 따라 apoptotic 세포가 유의적으로 증가하였으며, 특히 early apoptosis 보다 late apoptosis가 더 많이 진행됨을 확인할 수 있었다. 이를 바탕으로 MDA-MB-231 세포사멸과 관련된 유전자를 확인하고자 RT-PCR과 western blotting을 수행한 결과, 세포사멸의 주요한 조절인자인 anti-apoptotic인 bcl-2 발현이 두릅 추출물 처리 시 농도 의존적으로 감소되었고, 반대로 Bax의 발현은 증가됨을 확인하였다. 이를 통해 불활성화 형태로 존재한 pro-caspase-9과 pro-caspase-3의 발현이 시료 농도가 증가할수록 감소되었고, 활성화된 형태인 cleaved caspase-9과 cleaved caspase-3의 발현은 두릅 추출물 처리 농도에 의존적으로 증가하였다. 뿐만 아니라, 세포 사멸 과정의 주요 인자인 PARP 또한 시료 농도가 증가할수록 절단 현상이 비례적으로 증가하였다. 이상의 실험 결과를 근거로, 북한산 두릅 지상부의 70% ethyl alcohol 추출물이 MDA-MB-231 인체 유방암 세포의 증식을 억제하는 효과가 있음을 확인하였고, 세포사멸과 관련된 활성기전을 규명하였다. 추후 암 예방/치료제로서 두릅을 활용하기 위해서는 활성 본체와 안정성 규명 및 임상실험 등 추가 연구가 필요할 것으로 사료된다.

Terfenadine (TFN) was a second generation histamine receptor antagonist. Although several studies have reported the regulatory effect of H1-histamine receptor antagonists in human cancer cell lines, its effect in oral cancer remains unclear. In this study, we focused on addressing the anti-cancer activity of TFN in human oral cancer cell lines. The anti-cancer activities of TFN were performed by tryphan blue exclusion assay, 4'-6-diamidino-2-phenylindole (DAPI) staining, live/dead assay and Western blot analysis. TFN induced a significant reduction of the growth in three different human oral cancer cell lines (MC3, HSC4 and Ca9.22). TFN markedly induced apoptosis through DNA damage and increase in cytotoxicity. It also accumulated cleaved PARP and caspase 3. This process was due to cleavage of caspase 8 and Bid protein. The results from this study strongly demonstrated that the cleavages of caspase 8 and Bid are required for the apoptotic activity of TFN in human oral cancer cells. Taken together, these findings suggest TFN as a potent anticancer drug candidate for the treatment of oral cancer.

Epimedium koreanum Nakai has been used in traditional medicine as an aphrodisiac, hypotensive, and neurasthenia;however, the cellular mechanism of E. koreanum Nakai cultivated in North Korea on HCT116 colon cancer cellactivity has not been investigated. This study is to investigate the effect of E. koreanum Nakai on apoptosis of thehuman colon cancer cell line HCT116. The anti-proliferative activity of E. koreanum Nakai on HCT116 was iden-tified through MTT, Western Blot, and RT-PCR analyses. The results show that 70% ethyl alcohol extracts of E.koreanum Nakai inhibited the growth of HCT116 colon cancer cells (IC50: 1.2mg/mL on 24 hr). Concomitant acti-vation of the mitochondria-dependent apoptotic pathway occurred via modulation of Bcl-2 and Bax expressions,resulting in activation of cleaved caspase-3 and cleaved caspase-9.

Neuromedin B (NMB) acts as a growth factor or a morphogen and plays a role in cancer progression. Indeed, the NMB receptor (NMB-R) is overexpressed in different types of tumors. In our current study, we investigated the involvement of NMB-R in the proliferation of oral cancer cells. Human oral squamous cell carcinoma (SCC) and human oral cancer cells, SCC-25 cells were found to be NMB-R-positive. The NMB-R antagonist PD168368 inhibited the proliferation of SCC-25 cells and reduced their colony formation capacity. We also found that PD168368 induced the cell cycle arrest and apoptosis of SCC-25 cells in a dose-/time-dependent manner. Overall, this antitumor activity of PD168368 in human oral cancer cells suggests that NMB-R is a potential target for the future prevention and treatment of human cancers.

Dibenzylideneacetone (DBA), an analogue of curcumin has been shown to have anti-cancer activity in a variety of tumor cell lines. However, the anti-cancer activity of DBA and its molecular mechanism in HN22 oral cancer cell line have not been fully explored. The effects of DBA on anti-proliferative and apoptotic activity were evaluated by the trypan blue exclusion assay, 4’-6-diamidino-2-phenylindole (DAPI) staining, Western blot analysis, and reverse transcriptase-polymerase chain Reaction (RT-PCR). Our data showed that the treatment of DBA to HN22 cells exerted anti-proliferative and apoptotic activities and the activity was accompanied by a decrease in Sp1 protein, Sp1 mRNA and its promoter activity. DBA also reduced the expression level of Sp1 protein and caused apoptotic cell death in HN22 cells simultaneouly. Phosphorylation of ERK and JNK were regulated by DBA whereas phosphorylation of p38 was not altered. Overall, our results suggest that the regulation of Sp1 activities and ERK/JNK are involved in DBA-induced apoptosis and DBA can be a promising anticancer drug candidate for the treatment of oral cancer.

Colorectal cancer is the third most commonly diagnosed cancer in the world, nearly all patients diagnosed with this cancer die from it. Antibodies are glycoprotein molecules, which can efficiently recognize and eliminate specific pathogenic and disease antigens. Antibody researches for the last several decades have demonstrated the potential of therapeutic antibodies to fight cancer. Monoclonal antibody (mAb) CO17-1A recognizes the tumor-associated antigen GA733-2, a cell surface glycoprotein highly expressed in colorectal carcinoma cell, which is applicable for preventing and curing colorectal cancer. We have currently established baculovirus insect cell expression system to produce anti-colorectal cancer mAb CO17-1A. In this study, mAb CO17-1A was expressed in the transgenic insect cell line SWT4, which has humanized glycosylation processing pathway. Immunoblot confirmed that mAb CO17-1A properly expressed in SWT4. mAb CO17-1A was purified using protein G affinity column. In addition, Maldi-TOF verified that the mAb fused to KDEL, ER retention signal had high mannose type of glycan structure whereas the mAb without KDEL had partially humanized glycan structure. These results suggest that the insect cell expression system with the SWT4 possibly can be used as a useful alternative way to produce full-size mAb with humanized glycan structures for cancer immunotherapy.

Prostate cancer is a leading cause of death among the aging men. Surgical or radio therapy is effective when the cancer is confined to the prostate gland but once the cancer spreads beyond the pelvis, even chemotherapy and hormonal ablation therapy fails in curing this disease. Our previous studies have shown that β-glucan induces apoptosis in prostate cancer cells. Cellular viability of these cells treated with β-glucan was measured by MTT assay. β-glucan induced dose- and time-dependent inhibition of cell viability in LNCaP cancer cells. In flow cytometry analysis, β-glucan induced dose- and timedependent apoptotic activities in LNCaP cancer cells. In addition, increased of expression caspase-3, caspase-9, Bax, and cytochrome C but decreased of expression Bcl-2 was observed in LNCaP cells treated with β-glucan. These results suggest that β-glucan induces apoptosis in LNCaP human prostate cancer cells mediated mainly through the increased of expression caspase-3, -9, Bax, cytochrome C and decreased of expression Bcl-2.

Bile acids and synthetic bile acid derivatives induce apop-tosis in various kinds of cancer cells and thus have anti-cancer properties. Recently, it has been suggested that autop-hagy may play an important role in cancer therapy. How-ever, few data are available regarding the role of autophagy in oral cancers and there have been no reports of autophagic cell death in OSCCs (oral squamous cell carcinoma cells) in-duced by HS-1200, a synthetic bile acid derivative. We thus examine whether HS-1200 modulates autophagy in OSCCs. Our findings indicate that HS-1200 has anticancer effects in OSCCs, and we observed in these cells that autophagic vacuoles were visible by monodansylcadaverine (MDC)and acridine orange staining. When we analyzed HS-1200-treated OSCC cells for the presence of biochemical markers, we observed that this treatment directly affects the conversion of LC-3II, degradation of p62/SQSTM1 and full-length beclin-1, clea-vage of ATG5-12 and the activation of caspase. An autop-hagy inhibitor suppressed HS-1200-induced cell death in OSCCs, confirming that autophagy acts as a pro-death signal in these cells. Furthermore, HS-1200 shows anticancer acti-vity against OSCCs via both autophagy and apoptosis. Our current findings suggest that HS-1200 may potentially cont-ribute to oral cancer treatment and thus provide useful infor-mation for the future development of a new therapeutic agent.

비록 천연유래물질 중의 하나인 통초가 항산화작용, 항염증작용 및 해열작용과 같은 효능이 있다고 알려져있지 만, 종양에서의 암예방효능에 대한 연구는 보고된 바가 없 다. 본 연구에서는 사람 자궁경부암세포주인 HEP-2세포에서 통초메탄올추출물의 증식억제 효능 및 관련 분자표적을 확인하고자 하였다. 통초메탄올추출물은 세포증식을 유의성있게 억제하고, 세포사멸을 유도하는 것으로 나타났다. 또한, Bid의 발현에 영향을 주어 truncation을 유도하고, 이로 인해 cytochrome c가 마이토콘드리아에서 세포질로 이 동하도록 유도하였지만, Bid 이외의 다른 Bcl-2 family 에는 영향을 주지 못했다. 따라서, 이러한 결과를 종합해 볼 때, 통초메탄올추출물은 자궁경부암에서 암예방효능을 가진 잠재성있는 천연추출물이 될 수 있을 것으로 사료된다.

Considering the dearth of information regarding the medicinal properties of Luffa cylindrica, we assessed the antioxidative, antimutagenic and hyperplasia inhibitory activity of cancer cells from Luffa cylindrica extracts by employing biological and biochemical assays. Ethanol extracts of Luffa cylindrica inhibited MDA-BSA (malondialdehyde-bovine serum albumin) conjugation reaction (66.38±2.65), DPPH (1, 1-diphenyl-2-picryl-hydrazyl) radical production (60.13±0.42) and lipid peroxidation (56.04±3.24). In this study, Luffa cylindrica is believed to exert possible antioxidative effects. The direct and indirect antimutagenic effects of the ethanol extracts of Luffa cylindrica were examined by the Ames test using Salmonella typimurium TA98 and TA100. The inhibitory effects on indirect and direct mutagenicity shows an weak tendency, particularly in direct mutagenicity mediated by 2-nitrofluorene in Salmonella typimurium TA98 (5.82±5.74) and in indirect mutagenicity mediated by 2-anthramine in Salmonella typimurium TA100 (5.76±2.15). The ethanol extracts of Luffa cylindrica on cancer cell hyperplasia inhibitory activity via MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide) assay exerted cytotoxic effects on Hela cells (55.83±3.83) and MCF-7 cells (33.03±2.09), which were used in this study. Based on these results, it believed that the ethanol extracts of Luffa cylindrica have antioxidative capacities as well as hyperplasia inhibitory activity of cancer cells. Furthemore, Luffa cylindrica is a candidate for the prevention and dietetic treatment of chronic diseases and for the development of functional food.

MicroRNAs (miRNAs) are small non-coding RNAs that mediate gene expression at the post-transcriptional level by degrading or repressing targeted mRNAs. These molecules are about 21-25 nucleotides in length and exert their effects by binding to partially complementary sites in mRNAs, predominantly in the 3'-untranslated region (3'-UTR). Recent evidence has demonstrated that miRNAs can function as oncogenes or tumor suppressors through the modulation of multiple oncogenic cellular processes in cancer development, including initiation, cell proliferation, apoptosis, invasion and metastasis. In our present study, we examined the expression profile of miRNAs related to oral cancer cell growth inhibition using normal human oral keratinocytes (NHOK) and YD-38 human oral cancer cells. By miRNA microassay analysis, 40 and 31 miRNAs among the 1,769 examined were found to be up- and down-regulated in YD-38 cells compared with NHOK cells, respectively. Using qRT-PCR analysis, the expression levels of miR-30a and miR-1246 were found to be increased in YD-38 cells compared with NHOK cells, whereas miR-203 and miR-125a were observed to be decreased. Importantly, the overexpression of miR-203 and miR-125a significantly inhibited the growth of YD-38 cells. This finding and the microarray data indicate the involvement of specific miRNAs in the development and progression of oral cancer.

Endocrine disrupting chemicals (EDCs) have detrimental effects on human health. Among these EDCs, bisphenol A (BPA) binds to estrogen receptors (ERs) to stimulate estrogen-mediated responses. BPA is assumed to disrupt the reproductive and developmental system of humans. In addition, BPA has recently been suspected as a risk of carcinogenesis. Because BPA can cause abnormal estrogen-mediated response in the organism, exposure to BPA may stimulate growth of estrogen-dependent breast cancers in human. In breast cancer, cyclin E and cyclin-dependent kinase inhibitor p27 are important in G1/S phase transition during cell cycle progression. In this study, using an MTT assay, we investigated the effect of BPA on proliferation of MCF-7 breast cancer cells in vitro. In addition, we also analyzed the transcriptional levels of cyclin E and p27 following treatment with BPA using semi-quantitative RT-PCR. As a result, treatment with BPA resulted in significant induction of breast cancer cell growth, compared to a vehicle. BPA caused alterations of cyclin E and p27 mRNA expression. Expression of cyclin E was increased by BPA, while p27 was decreased at 24 h after treatment with BPA in MCF-7 breast cancer cells. Taken together, these collective results suggest that exposure to BPA induced breast cancer cell proliferation with deregulation of the cell cycle. A further study is required in order to determine the effects of BPA on the carcinogenic process in in vivo models.

Betulinic acid (BA), a naturally occurring triterpene found in the bark of the white birch tree, has been investigated to induce apoptosis in various cancer cells and animal models. However, there is no report of the chemopreventive effect of BA in cervical cancer cells. Using KB human cervical cancer cells as a model, we currently show that BA decreases cell viability and induces apoptotic cell death. The mechanism of the BA-induced anti-growth response in KB cells is due to the down-regulation of specificity protein 1 (Sp1) and its downstream targets, myeloid cell leukemia-1(Mcl-1) and survivin. Thus, BA acts as a novel chemopreventive agent through the regulation of Sp1that is highly expressed in tumors.

Cancer cells are often found in an ischemic condition due to the rapid outgrowth of their vascular supply, and these cells are expected to develop an increased potential for local invasive growth. Since the first steps are characterized by increased motility and invasiveness, expression of molecules involved in cellular adhesion to extracellular matrix (ECM) is increased in the process of cancer cell invasion and metastasis. In this work we explored the molecular characteristics and its regulatory mechanism of hypoxic oral squamous cell carcinoma (OSCC) cells. Our experiment identified that hypoxia increases α5 integrin protein levels through phosphoinositide 3-kinase (PI3K)/Akt pathway in OSCC cells.