The objective of this study was to investigate the anticancer effects of EMPS (edible mushroom mycelium polysaccharide: Tremella fuciformis) in animal models with colorectal cancer induced by AOM/DSS. The experimental groups consisted of Nor (normal), NC (AOM/DSS), EMPS (EMPS 50, EMPS 100), and PC (Fluorouracil). The NC group had the highest number of colon tumors, whereas it was observed that tumor occurrence was significantly reduced in the EMPS consumption group. The expression of Bcl-2, an apoptosis inhibitor, was significantly lower in the EMPS 50 & 100 and PC groups. On the other hand, the mRNA gene expression of Bax, a factor that induces apoptosis, was significantly higher in the EMPS 50 & 100 and PC groups compared to the NC group. The mRNA expression levels of TNF-α and COX-2 significantly increased in the NC group, but showed a significant decrease in the EMPS and PC groups, indicating inhibition of the cancer-promoting response of cells. At the phylum level of the mice's intestinal microbial composition, the proportion of Bacteroidetes tended to decrease, while the proportion of Firmicutes tended to increase with EMPS administration. This suggests that changes in the gut microbiota caused by inflammation can be influenced by dietary intake.

Vinpocetine induces anti-inflammatory effects in various inflammatory diseases via the inhibition of phosphodiesterase type-1-independent nuclear factor-κB signaling pathway and the release of inflammatory cytokines. In this study, we investigated the effect of vinpocetine on the proliferation of colon cancer cells and its underlying molecular mechanisms. Our data showed that vinpocetine inhibits the viability and proliferation of colon cancer cells. Vinpocetine treatment induced cell death in HCT116 cells, which the percentages of sub-G1 phase were significantly increased, and the apoptosis-related genes were regulated after HCT116 cells were treated with vinpocetine. In sum, our findings indicated that vinpocetine could be a therapeutically useful candidate in the treatment of colon cancer.

Purpose: The objective of this study was conducted to investigate the effects of rutin, buckwheat components on cell growth and anti-inflammation in adipocyte 3T3-L1 and human colon cancer cell SW-480. Methods: We cultured 3T3-L1 adipocyte and SW-480 colon cancer cell to confluence, at which time starvation was induced with SFM for 1 day. Cells were then cultured in medium containing 0, 25, 50, or 100 μmol/mL of rutin 3T3-L1 or 0, 10, 20, or 40 μmol/mL SW-480. Cell viability was measured using a cell viability kit. In addition, we examined the expression of mRNA related to inflammation. RT-PCR was used to quantity tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) mRNA levels. Results: Rutin significantly inhibited 3T3-L1 and SW-480 cell proliferation in a dose and time dependent manner. Rutin also significantly reduced the mRNA expression of IL-1β, IL-6 and TNF-α at the highest dose. In addition, rutin treatment caused a significant reduction in COX-2 and iNOS mRNA levels compared to the control group. Conclusion: Overall, our results suggest that rutin has the potential to reduce inflammation, and that these effects are greater during tissue-damaging inflammatory conditions.

Colorectal cancer is a major cause of morbidity and mortality that accounts for over 9% of all incidences of cancer. Additionally, colorectal cancer is widely recognized as an environmental disease related to ill-defined cultural, social and lifestyle factors including physical activity, obesity, cigarette smoking and heavy alcohol consumption. Accordingly, natural phytochemicals and extracts have attracted attention because of their beneficial biological effects. Coenzyme Q10 (CoQ10) is a common supplementary medicine applied to increase bioenergetic capacity in various diseases. Therefore, in this study, we investigated whether CoQ10 treatment has any inhibitory effects and its related cellular mechanisms in human colon cancer HCT116 cells. A MTT assay revealed that CoQ10 slightly decreased the proliferation of HCT116 cells; however, glutathione- and superoxide dismutase- activity were unchanged in response to CoQ10 treatment. A DCF-DA assay revealed that CoQ10 slightly increased ROS release of HCT116 cells. However, in a nitric oxide (NO) assay, CoQ10 significantly increased NO production in a dose-dependent manner. The results of western blot analysis revealed that the protein levels of Bax, p21 and p53 were increased, whereas the protein level of Bcl2 was decreased suggesting that the CoQ10-mediated inhibitory mechanism is associated with apoptotic signaling. Taken together, our findings indicate that CoQ10 has an inhibitory effect on the growth of colon cancer cells via NO production that is associated with regulation of factors involved in apoptotic signaling including Bax, Bcl2, p21 and p53.

해양생물을 포함한 천연물질은 신약개발의 원천 소재로서 매력적이며, 특히 무수한 미지의 해양생물들의 연구가 관심을 받고 있다. 기존의 연구에서 미크로네시아에서 채취한 해면동물 40여종에 대하여 항증식 효과를 다양한 암세포주에서 검색한 바 있다. 본 연구에서는 그 중 Cos-cinoderma sp.의 작용 및 그 기전을 살펴보았다. 특히, 암억제유전자 p53의 발현을 억제시킨 세포주(HCT116 p53KO 과 RKO-E6)에서의 차이점을 비교하였다. 세포생존률 시험에서 Coscinoderma sp. 추출물은 p53의 유무와 상관없이 암세포의 증식을 억제하였음을 확인하였다. 이 암세포증식 억제 효과가 p53 존재에 따라 다르게 나타나는지 알아보기 위하여 세포사멸 관련 단백질 발현양을 Coscinoderma sp. 처리한 각 세포주에서 비교하였다. 그 결과, Coscinoderma sp.를 HCT16 세포주에 처리하였을 때, p53과 Noxa의 발현이 증가하는 것을 관찰하였고, caspase-9이 분절되면서 감소하는 것으로부터 apoptosis를 일으킨다고 여겨진다. 반면, p53이 결핍된 HCT116세포주에서는 Coscinoderma sp. 에 의하여 p21과 mTOR의 발현이 증가되는 것을 확인하였고, 이는 senescence를 야기할 수 있다고 여겨진다. 본 연구로부터 Coscinoderma sp.는 p53의 존재여부에 따라 상이한 작용기전을 매개하여 대장암 세포주의 증식을 억제한다는 것을 알 수 있었다. 이는 새로운 항암제의 개발 가능성을 제시하는 것으로, Coscinoderma sp.의 활성 성분에 대한 지속적인 연구가 이루어 져야 할 것으로 보인다.

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.

The objective of this study was to determine the effect of macrophages on growth of human colon cancer cells. The results showed that co-culture of colon cancer cells with macrophages inhibited the growth of colon cancer cells (HCT116 and SW620) depending on the number of macrophages, RAW 264.7 cells, and activated THP-1 cells accompanied by down regulation of pSTAT3 in cancer cells. We also found that expression and release of cancer cell growth inhibitory cytokines, IL-1 receptor antagonist (IL-1ra) and IL-10, was increased in macrophages. Blocking of the STAT3 pathway with specific inhibitor and siRNA of STAT3 abolished the growth of colon cancer cells and expression of IL-1ra and IL-10. In addition, neutralization of IL-1ra and IL-10 with antibodies resulted in reversal of macrophage-induced inhibition of cancer cell growth. These data showed that IL-1ra and IL-10 released from macrophages inhibit growth of colon cancer cells through inhibition of the STAT3 pathway.

본 실험에서는 대장암 세포 HT-29의 증식을 억제하고 세포 사멸을 유도하는 천연소재 발굴을 목적으로, 오미자(Schizandra chinensis Baillon) 열수 추출물을 이용하여 인체 대장암 세포 HT-29의증식에 미치는 영향을 확인하였다. 오미자 열수 추출물이 HT-29 대장암 세포의 apoptosis 유도 효과 및 기전에 미치는 영향을 분자생물학적 방법으로 실험하여 다음과 같은 결론을 얻었다. MTT assay를 통해 인체 대장암세포 HT-29는 오미자 시료농도 0, 1.0, 2.0, 4.0 mg/mL에서 암세포 사멸농도가 각각 0%, 10%, 70%, 88%를 나타내었다. 대장암세포에 오미자 추출물을 처리하고 cell cycle 분석 결과, 시료농도 의존적으로 sub-G1기가 증가하였고, G0/G1기는 감소되는 것을 통해, apoptosis가 일어나 세포 증식을 저해하는 것으로 확인되었다. 대장암세포 핵의 형태학적 변화를 보면, 오미자 추출물 처리 시 농도 의존적으로 세포수가 감소되는 것이 뚜렷이 관찰되었고 cell shrinking, chromatin condensation 등 apototic body 등과 같은 형태학적 변화들이 뚜렷하게 관찰되었다. RT- PCR을 통한 유전자 발현은, 오미자 추출물 농도 의존적으로 p53 유전자 발현이 증가되는 것을 통해 대장암세포의 증식억제를 확인할 수 있었다. In vitro 실험에서 오미자 열수추출물이 대장암세포의 성장을 저해하는 효과가 있음을 확인하였으며, 오미자 추출물에 함유된 활성 본체 규명 및 apoptosis를 유도하는 작용기작에 관한 연구가 수행중이다.

Radiotherapy is one of the major therapies for cancer treatment. p53 acts as a central mediator of the cellular response to stressful stimuli, such as radiation. Recently it has been known that activation of the phosphatidylinositol- 3-kinase (PI3K) pathway is associated with radioresistance. In this study, we investigated whether X-irradiation up-regulates PI3K in a p53-dependent manner in human colon cancer cells. In order to study this phenomenon, we have treated p53-wild type and p53-mutant type HCT116 cells with X-ray. Treatment of wild type HCT116 cells with 8 Gy resulted in a marked increase in PI3K (p85), which paralleled an increase in PTEN, a counterpart of PI3K. However, these effects of X-rays in the p53-mutant cells were not observed. These results suggest that the X-irradiation- induced up-regulation of PI3K/PTEN pathway is p53-dependent.

Chronic inflammatory diseases such as Crohn′s disease and ulcerative colitis are associated with increased risk of colon adenocarcinoma. Apoptic induction of colon cancer cells by cytokines and death receptors is an important anti-cancer therapy. We observed that co-administration of TNFα and IFNγ in human colon cancer cell line, HCT116, resulted in cell death and expression of IL-32. Cleavage forms of caspase-3, caspase-9, and PARP were increased in TNFα / IFNγ-treated HCT116. mRNA expression of death receptors, including TNFR1 and Fas were not changed and NO generation was not induced by combination of TNFα and IFNγ. However, mRNA expression of IL-32α, β, and γ was increased in TNFα / IFNγ-treated HCT116. To determine the effect of IL-32 in HCT116 cell apoptosis by TNFα / IFNγ stimulation, IL-32 siRNA-transfected HCT116 cells were cultured with TNFα / IFNγ and cell proliferation was measured. IL-32 siRNA induced slight recovery of cell viability of TNFα / IFNγ-stimulated HCT116. These results suggest that IL-32 is not directly related to apoptosis of HCT116 by TNFα / IFNγ stimulation. However, IL-32 expression by TNFα or TNFα / IFNγ in a colon cancer cell line is very interesting because of the unknown effect of IL-32 in colon cancer. Our study will contribute to development of studies for IL-32 function in human colon cancer and anti-cancer therapies using cytokines.

The Maillard Reaction Products (MRPs) such as Glucose-tyrosine (Glu-Tyr) and Xylose-arginine (Xyl-Arg) have antioxidant, antimutagenic, and antibacterial effects. However, to date, still little is known about the other biological effects of the MRPs. In this study, we investigated whether the fructose-tyrosine MRP, 2,4-bis(p-hydroxyphenyl)-2-butenal (Fru-Tyr), could modulate cell cycle progression and NF-κB activity, and thereby induce apoptotic cell death of colon cancer cells. Treatment with different concentrations (10-40 μg/ ml) of Fru-Tyr for 24 h inhibited colon cancer cell (SW620 and HCT116) growth followed by induction of G₂/M phase cell cycle arrest and apoptosis in a dose-dependent manner. We also found that Fru-Tyr suppressed tumor necrosis factor-alpha (TNF-α)-induced NF-κB transcriptional activity. Moreover, Fru-Tyr induced the expression of apoptotic gene, cleaved caspse-3. These results suggest that Fru-Tyr inhibited colon cancer cell growth through induction of G₂/M phase cell cycle arrest and apoptotic cell death by modulating of NF-κB.

The HMG box containing protein (HBP) has a high mobility group domain and involved in the regulation of proliferation and differentiation of tissues. We screened HBP2 in glioblastoma using Suppression Subtractive Hybridization (SSH) and isolated human spermatogonial stem cell‐like cells (hSSC‐like cells) derived from patients of nonobstructive azoospermia (NOA). Expression of HBP2 was analyzed by RT‐PCR in undifferentiated stem cells (human Embryonic Stem Cells, hSSC‐like cells 2P) and spontaneous differentiated stem cells (hSSC‐like cells 4P). It was overexpressed in hESC and hSSC‐like cells 2P but not in hSSC‐like cells 4P. Also, the expression level of HBP2 was downregulated in colon tumor tissues compared to normal tissues. Specifically in synchronized WI‐38 cells, HBP2 was highly upregulated until the G1 phase of the cell cycle and gradually decreased during the S phase. Our results suggest that HBP2 was downregulated during the spontaneous differentiation of hSSC‐like cells. HBP2 was differently expressed in colon tissues and was related to G1‐progression in WI‐38 cells. It may play a role in the maintenance of an undifferentiated hSSC‐like cell state and transits from G1 to S in WI‐38 cells. This research was important that it identified a biomarker for an undifferentiated state of hSSC‐like cells and characterized its involvement to arrest during cell cycle in colon cancer.

Nerve gro때h factor-induced B (NGFI-B, Nur77) is an orphan nuclear receptor with no known endogenous Iigands , however‘ recent stuclies on a series of methylene -substituted diindolylmethanes (C-DIMs) have identified 1,l-bis(3’ - In dolyl) -l-(phenyl)methane (DIM-C-Ph) and l , l -bis(3’ indolyl)-l-(p-anisyl)methane (DIM-C-pPhOCHa) as Nur77 agonist Nur77 is expressed in several colon cancer cell lines (RKO, SW480, HCT-116, HT-29 and HCT-15) and we a lso observed by irnmunostaining that Nur77 was overexpressed in colon tumors compared to normal colon tIssue DIM-C-Ph and DlM-C-pPhOCH3 decreased survival and induced apoptosis in RKO colon cancer cells and this was accompanied by in ductdion of tumor nec rosis factor-related apoptosis-incluced ligand (TRAlL) protein, The induct ion of a poptosis and TRAlL by DIM-C-pPhOCH3 was significantly inhibited by a small inhibitory RNA for Nur77 (iNur77); however, it was evide nt from RNA in terference studies that DIM-C-pPhOCH3 a1so induced Nur77-independent apoptosis. Analysis o( DIM-C-pPhOCH3-induced gene expression using microarrays idontifiod sovoral proapoptotic genos and analysis by ro verse t ranscriptase PCR in the presence 0 1' absence of iNru77 showed that incluction of prograrnmed cell death gene 1 (PDcm) was Nur77-dependent‘ whereas induction of cystathionase (CSE) and activating transcription factor 3 (ATF3) was Nur77-independent, DIM-C-pPhOCHa (25 mg/kg/day) also inhi bited tumor growth in athymic nude mice bearing RKO cell xenograft, These results demonstrate that Nur77-active C-DIM compounds represent a new class of anti-colon cancer drugs that act through receptor- dependent and - independent pathway

1,1- Bis(3’-indolyl)-l-(p-methoxyphenyl)methane (DIM- C- pPhOCH.3) is a methylenc - substituted diindolylmethanes (C-DIM) ana log that acti vates the orphan receptOl‘ nerve growth factor-induced-B (NGFI-B, Nur77) , RNA inteference studies with small inhibitory RNA for Nur77 demonstrate that DIM-C-pPhOCH:J induces Nur77-dependent and - independent apoptosis, and this study has focused on delineating the Nur77-independent proapoptotic pathways induced by the C-DIM analog DIM-C-pPhOCH3 induced caspase-dependent apoptosis in RKO colon cancer cells through decreased mitochondrial membrane potential which is accompanied by increased mitochondrial bax/bcl-2 ratios and release of cytochrome c into the cytosol DlM-C一pPhOCH.3 also induced phosphatidylinositol-3-kinase-dependent activation of early growth response gene-l whi ch, in turn, induced expression of the proapoptotic nonsteroidal anti-inflammatory drug- activated gene-l (NAG- l) in colon tumors in athyrnic nude mice bearing RKO cells as xenografts, DIM-C-pPhOCH.3 also activated the extrinsic apoptosis pathway through increased phosphorylation of c- jun N-terminal kinase which, in turn, activated C/EBP homologous transcription factor (CHOP) and death receptor 5 (DR5) , Thus, the effectiveness of DIM-C-pPhOCH.3 as a tumor growth inhibitor is through activation of Nur77-dependent and -independent pathways

Backgrounds : The action mechanisms of several chemopreventive agents derived from herbal medicine and edible plants have become attractive issues in cancer research. Tea is the most widely consumed beverage worldwide. It have been demonstrated that the active principles of tea sources such as flower extract Camellia sinensis (CSF) and Camellia japonica (CJF)were attributed to their tea polyphenols. We focused on investigating CSF, CJF, mixtures of CSF and CJF has been proven to suppress colonic tumorigenesis. Methods and Results : In this study, human colorectal carcinoma HT-29 cells were treated with CSF, CJF, mixture of CSF and CJF to examine the anti-proliferative and pro-apoptotic effects of mixture of CSF and CJF (3 : 1), as well as the molecular mechanism underlying these effects. Cell viability assay, nuclear staining, DNA fragmentation, caspase assay, cytochrome c release, were utilized to dissect the signaling pathways. In mixture of CSF and CJF (3 : 1), CSF appeared most anticancer effect by both MTT assays and the cleavage analysis of apoptosis-related molecules and PARP. Interestingly, we found that CJF make it possible to express the apotosis inducing by CSF in a short time and apoptosis effect of CSF maintained sustainable. Conclusion : In summary, our results from this study suggest that in HT-29 human colon cancer cells (i) CSF treatment causes damage to mitochondria, and (ii) CJF contributed CSF induced apoptotic cell death mediates cytochrome C release, (ⅲ) mixture of CSF and CJF (3 : 1) the potential to function as a chemopreventive agent against colon cancer.

Background : The Aralia cordata and Dendropanax morbifera, belonging to the family Araliacea, is a perennial herbaceous species. Although its beneficial effects for several chronic diseases are well-known, the role of its effects on chronic intestinal inflammation, colon carcinogenis are limited. Inflammatory bowel diseases (IBD) are chronic inflammatory disorders of gastrointestinal tract. It has been reported that IBD is associated with colon cancer which is one of the most common types of cancer diagnosed and a major cause of cancer-related mortality. Methods and Results : The present study hypothesized that Araliacea family would exert a protective effect on inflammation and dextran sulfate sodium (DSS)-induced colitis mouse model. Effect of Aralia cordata extracts (ACE) and Dendropanax morbifera leaf extracts (DMLE) on pro-inflammatory markers, mitogen-activated protein kinase signaling (MAPKs), activation of nuclear factor κB (NF- κB) in THP-1 human monocyte and HT-29 colon epithelial cell model were investigated. In addition, colon inflammation inhibitory effects of ACE and DMLE on the characteristics of in DSS-induced colitis model. Results showed that ACE, DMLE attenuated the severity of DSS-induced colitis, as assessed by disease activity index scores, shrinkage of colon length, and histopathologic changes and infirmatory cytokine concentration. Furthermore, ACE treatment significantly suppressed the colon carcinogenesis in AOM/DSS induced colon cancer model. The apoptosis induction abilities of the DMLE were studied by analyzing the expressions of Bcl2 family protein, caspase, Annexin V/PI staining and DNA fragmentation. According to our results, DMLE significantly induced cell death in SW-480 cells. Apoptosis was determined by cell morphology and electrophoresis of DNA fragmentations. Furthermore, treatment with DMLE also induced the increase in caspase activity, pro-apoptotic proteins (Bax and Bad) and the decrease in anti-apoptotic proteins (Bcl-2 and Bcl-xL). In addition, DMLE induce cell cycle arrested at the G1/S phase in SW-480 cells and exhibited significant inhibition on the expressions of G1/S phase specific CDK, cyclin and up regulation of CDK inhibitors. Conclusion : Taken together, these results provide evidence that ACE and DMLE prevention colon cancer by regulating inflammation, colitis, and colon carcinogenesis which indicates its benefit for colon health.

본 연구는 폐기되고 있는 대두박을 기능성 식품 원료로 이용하기 위하여 대두박 70%에탄올 추출물로부터 Diaion HP-20 흡착수지를 이용하여 조사포닌을 분리 한 후 이들에 대한 항산화 효과 및 대장암세포 성장 억제효과를 조사하였다. Diaion HP-20 흡착 수지를 이용하여 분획한 물, 20% 및 100% 주정 분획물을 TLC상 확인한 결과 100% 주정분획물에서 조사포닌들이 함유되어 있음을 확인 할 수 있었다. 대두박 조사포닌은 고농도에