In this study, the quality characteristics of kimchi, such as its salinity, pH, and acidity, were measured and compared, and the HT-29 human colon cancer cells were used to show the anticancer effects of kimchi. The kimchi samples used herein included standard kimchi (SK), turnip kimchi (TK), and turnip-powder-added kimchi (TPK). The measured pH and acidity of TK and TPK showed no significant differences with those of SK. Compared to SK and TK, TPK had higher DPPH scavenging activity and higher total flavonoid content, confirming its antioxidant activity. The cancer cell growth inhibition rates of TK and TPK were significantly higher than that of SK. In HT-29 cells treated with TPK, the mRNA expression of Bcl-xL, an anti-apoptosis-related gene, was lower, and the mRNA expressions of the apoptosis-related genes Bax, Bad, and caspase-9 were higher. TPK showed significantly higher levels of mRNA expressions for the cell-cycle-related genes p53 and p21 than the other samples, in addition to suppression effects on cancer cell proliferation. Compared to SK, TK and TPK suppressed the growth of colon cancer cells and showed higher anticancer effects. Therefore, it is shown that kimchi with added turnip powder had high anticancer effects.

The purpose of this study was to investigate the biological activity of fucoidan, a sulfur-containing polysaccharide, on cytotoxicity and apoptosis in the human HT-29 colorectal cancer cell line using cell viability, Flow cytometry, Western blot, and RT-PCR analyses. Fucoidan inhibited the proliferation of HT-29 cells by 39.6% at a concentration of 100 μg/mL for 72 h. The inhibition was dose-dependent and accompanied by apoptosis. Flow cytometric analysis showed that fucoidan increased early apoptosis and late apoptosis by 65.84% and 72.09% at concentrations of 25 and 100 μg/mL, respectively. Analysis of the mechanism of these events indicated that fucoidan-treated cells exhibited increases in the activation of caspase-3, caspase-8, and PARP in a dose-dependent manner. These results suggest that fucoidan may inhibit the growth of human colorectal cancer cells by various apoptosis-promoting effects, as well as by apoptosis itself.

Lindera glauca Blume has been used in Korean traditional medicine to treat the symptoms of paralysis, abdominal pain, speech disorders, extravasations, contusions, and pain caused by rheumatoid arthritis. We investigated the effect of L. glauca Blume extracts on the proliferation of colorectal cancer cells in vitro using HCT116 human colorectal cancer cell lines. We also investigated its mechanism of action. For this purpose, we used the MTT assay, western blotting, DNA fragmentation analysis, and flow cytometry. HCT116 cells were cultured in several concentrations of ethanol extracts of L. glauca Blume root (0, 50, 100 μg/mL). In this study, colon cancer cell growth was inhibited by L. glauca Blume root extract in a dose-dependent manner. It was associated with induction of apoptosis as assessed by nuclear fragmentation and cell cycle analysis. Apoptosis was assessed using western blotting for TNF-α, IL-6, NF-κB, Caspase-3, PARP, Bax, Bcl-2, and SIRT1. The extract also dose-dependently upregulated the expression Bax, the pro-apoptotic gene and downregulated the expression of the anti-apoptotic gene Bcl-2. Furthermore, the extract enhanced Caspase-3 activity in a dose-dependent manner. Our findings provide evidence that L. glauca Blume extract may mediate its anti-proliferative effect via the modulation of apoptosis.

Colon cancer is one of the most common malignant tumors, but there are still a few validated biomarkers of colon cancer. Exosome-mediated microRNAs (miRNAs) have been recognized as potential biomarkers in cancers, and miRNAs can regulate a variety of genes. Recently, Fusobacterium nucleatum was discovered in the tissues of human colon cancer patients. Its role in colon cancer was highlighted. F. nucleatum may contribute to the progression of colon cancer through the mechanism of exosome-mediated miRNAs transfer. However, the exosomal miRNAs regulation mechanism by F. nucleatum in colon cancer is not well known. Thus, we performed next-generation sequencing to investigate the overall pattern of exosomal miRNAs expression in the colon cancer cell culture supernatant. We have confirmed the alterations of various exosomal miRNAs. In addition, to investigate the function of exosomal miRNAs, a Kyoto Encyclopedia of Genes and Genomes analysis was performed on the target genes of changed miRNAs. Potential target genes were associated with a variety of signaling pathways, and one of these pathways was related to colorectal cancer. These findings suggested that F. nucleatum can alter exosomal miRNAs released from colorectal cancer cells. Furthermore, exosomal miRNAs altered by F. nucleatum could be potential biomarkers for the diagnosis and therapy of colon cancer.

Aloin [1,8-Dihydroxy-10-(β-D-glucopyranosyl)-3- (hydroxymethyl)-9(10H)-anthracenone]은 알로에에서 추출한 천연 안트라퀴논이다. 다양한 유형의 인간 암세포에서 항산화, 항암 효과가 있는 것으로 밝혀졌지만 인간 대장암 세포 HT-29에서 aloin의 항암 효과는 밝혀지지 않았다. 본 연구에서는 aloin이 인간 대장암 HT-29 세포에서 세포 사멸 작용을 발휘할 수 있는 메커니즘을 조사하였다. Aloin 이 세포 생존율에 영향을 미치는지 알아보기 위해 대장암 세포 HT-29, 흑색종 세포 A375SM, 위암 세포 AGS를 aloin(0, 100, 200, 300 및 400 μM)으로 처리하였을 때, HT- 29에서는 농도 의존적으로 세포 생존율을 감소시켰고, A375SM과 AGS 세포에서는 암세포 생존율의 감소가 보이지 않았다. 이러한 HT-29에서의 세포 생존율 감소가 세포자멸사로 인한 감소인지 확인하기 위해 DAPI stain과 flow cytometry를 실시한 결과 apoptotic body가 유의적으로 증가하고 세포 자멸사가 증가한 것을 확인할 수 있었다. 이와 같은 결과를 바탕으로 aloin이 대장암 세포 HT- 29에서 세포 사멸 관련 단백질 발현 양상에 미치는 영향을 확인하기 위해 western blotting을 실시하였다. Aloin은 Bax, PARP의 분절을 농도 의존적으로 증가시켰고, caspase- 3, -8을 활성화시켰지만, Bcl-2는 대조군에 비해 변화가 없었다. Aloin에 의해 유도된 세포자멸사 기전을 확인하기 위해 MAPK pathway 중 p-p38과 p-ERK의 발현을 확인한 결과, p-p38을 up-regulation시키고 p-ERK의 downregulation을 유도했다. 따라서, aloin은 인간 대장암에서 암 세포 성장 억제 효과 및 암세포 사멸 유도로 암예방 약제로서의 개발 가능성이 있다고 사료된다.

L-carnitine은 라이신과 메티오닌으로 생합성되며 골격근 과 심근을 포함한 다양한 동물조직에서 발견된다. L-carnitine이 포함된 식품으로는 양고기, 소고기, 돼지고기 등이 있고 근육발달에 도움을 주며 뼈를 강화하거나 대사작용을 도와주는 기능을 하여 영양 보조제로 많이 섭취하는 것으로 알려져 있다. 최근 L-carnitine은 제 2형 당뇨병, 골다 공증, 대사성 신경증후군 등의 다양한 질병의 약물로도 연구 되고 있으며 암에서는 치료 보조제로 개발되어있다. 하지만 대장암에서의 L-carnitine에 대한 효과 및 기전에 대해서는 명확하지 않고 연구된 바가 없기 때문에 본 연구에서 저자들은 L-carnitine의 효능을 인간대장암세포주 HCT116에서 규명하고자 하였다. L-carnitine은 세포 내 활성산소종 (ROS)를 높은 수준으로 증가시켜 세포 증식을 억제하였다. 또한, 세포 증식과 죽음에 관련한 단백질 ERK1/2와 p38을 유의적으로 활성화 시킨다는 것을 입증 하였다. 이때, ERK1/2 억제제(PD98059)를 처치하여 ERK1/ 2의 활성화가 활성산소종 발생 및 세포사멸에 중요하다는 것을 밝혔다. 따라서, 본 연구 결과는 L-carnitine이 대장 암세포주의 증식을 억제 할 수 있고 이는 대장암의 치료에 있어 잠재적인 치료 물질이 될 수 있음을 시사하며 이 과정에 관여하는 신호전달기전을 조사하여 항암의 치료기 전에서 활성산소종이나 ERK1/2, p38 단백질의 활성화의 중요성을 제시하였다.

Recently, the area of marine resources has become concerned with sources for the next generation of the bio-industry. Until present, development of the marine resources has remained limited, although a large number of these resources are considered to have potential for various significant biological activities. Most marine sponges, marine algae and coral could be used to create specific compounds for survival against a harsh environment. Therefore, it was necessary that these materials needed to be elucidated with biological activities, such as like anti-inflammatory, anti-viral or anti-cancer effects for their utilization in the bio-industry. In this study, we screened extracts of marine resources for their anti-cancer effect on human colorectal cancer cells. These resources were collected at Kosrae of Micronesia on April, 2013 and extracted with methanol. Cytotoxicity of marine resources was observed. Of a total of 20 specimens, three specimens dose-dependently demonstration inhibition of cell viability. Furthermore, cells treated with these specimens for 48h were induced p53, p21, Bax and caspase-3. The results suggest that they involved p53-mediated apoptosis. Two positive specimens (1304KO-327 and 1304KO-329) were verified as the identical materials, which are Hyrtios sp. Unfortunately 1304KO-207 was not yet classified and needed to identify in the further study. There results suggested that marine resources with positive potential in anticancer effect would be good candidates as useful bio-resources.

The antigen GA733 is a cell-surface highly expressed glycoprotein on most human colorectal carcinomas. GA733 can be characterized as a cancer vaccine. In this study, GA733 was fused to the human immunoglobulin IgG Fc fragment to become recombinant gene GA733-Fc. Based on this, 4 recombinant genes were constructed as follows: GA733-Fc with signal peptide sequence and fusion of ER retention sequence (KDEL) (spGA733-FcK), GA733-Fc with signal sequence (spGA733-Fc), GA733-Fc fused to ER retention sequence (GA733-FcK) without signal peptide and GA733-Fc without signal peptide. Baculovirus-insect cell expression system is widely used for the high level production of recombinant proteins especially for glycoproteins. Constructed 4 recombinant genes were cloned to baculovirus express vectors. DH10Bac E.coli.-mediated transformation was used to generate recombinant bacmid DNA. Recombinant DNA was confirmed by PCR. Insect cell was transfected by bacmid to produce the recombinant baculovirus infects insect cell to produce recombinant protein. Western blot and sandwich ELISA showed the expression of recombinant proteins. Each cell lines (sf9 and HighFive) differed in recombinant proteins production level and protein secretion capability. N-Glycosylation analysis showed the function of signal peptide and ER retention sequence (KDEL). Taken together, baculovirus-insect cell system can be used to express recombinant GA733-Fc glycoproteins.

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.

The deleted in colorectal cancer (DCC) protein mediates attractant responses to netrin during axonogenesis. In the rat trigeminal ganglia (TG), axons must extend toward and grow into the trigeminal nerve to innervate target tissues such as dental pulp. Our present study aimed to investigate the exp- ression of DCC in the TG. Four developmental timepoints were assessed in the experiments: postnatal days 0, 7 and 10 and adulthood. RT-PCR and western blotting revealed that the expression of DCC mRNA and protein does not signi- ficantly change throughout development. Immunohistoche- mistry demonstrated that DCC expression in the TG was detectable in the perikarya region of the ganglion cells du- ring development. Nerve injury at 3 and 5 days after the man- dibular nerve had been cut did not induce altered expression of DCC mRNA in the TG. Moreover, DCC-positive cell bodies also showed similar immunoreactive patterns after a nerve cut injury. The results of this study suggest that DCC constituti- vely participates in an axonogenesis attractant in ways other than expression regulation.

본 연구 결과들을 바탕으로 도깨비부채 잎(RPL)은 GSK3β 활 성화를 통해 IκB-α를 인산화시켜 단백질 분해를 유도하고 Iκ B-α 분해로 인해 p65 핵내 전이를 유도하여 NF-κB 신호전달을 활성화?시킨다. 이러한 NF-κB 신호전달 활성화를 통해 대장암 의 세포생육을 억제하는 것으로 추정된다. 본 결과는 도깨비부채 잎을 소재로 항암을 목적으로 한 천연치료제 및 대체보완소재 개발에 활용할 수 있다고 판단된다. 그러나 도깨비부채 잎의 대장암에 대한 세포생육 억제와 작용기전의 정확한 관련성과 세포생육 억제활성 물질 분석을 위해 추가적인 연구가 필요할 것으로 사료된다.

본 연구에서 상동나무 가지 추출물(STB-E100)은 대장암 세포에서 세포사멸을 유도하여 세포생육을 억제하였다. 또한 Iκ B-α 인산화를 통한 IκB-α 단백질 분해를 유도하며 이로 인해 P65 핵내 전이를 유도하여 NF-κB 신호전달을 활성화시킨다. NF-κB 신호전달 활성화는 GSK3β 활성화를 통해 P65 핵내 전 이를 유도에 의한 것이지만 IκB-α분해는 GSK3β 의존성이 아니다. 상동나무 가지 추출물은 이러한 신호전달 활성화를 통해 세포사멸을 유도하여 대장암의 세포생육을 억제한다. 본 결과를 바탕으로 상동나무 가지가 암 예방 및 치료를 목적으로한 표적 요법에서 항암제 개발의 잠재적 활용 소재로서 이용 가능하다고 사료된다. 그러나 대장암 세포에서 상동나무 가지 추출물에 의해 유도된 NF-κB 신호전달 작용기전을 좀 더 구체적으로 구명할 필요가 있고 대장암에 대한 세포사멸과 작용기전의 정확한 관련성을 조사하기 위해 추가적인 연구가 필요하다.

이상의 연구 결과로 먹넌출 열매 추출물은 GSK3β 의존성 Cyclin D1 단백질의 분해를 통해 대장암세포의 생육 억제와 관련이 있는 것으로 확인된다. 본 결과는 대장암의 항암제 개발을 위한 소재로 먹넌출 열매의 활용이 가능할 것으로 판단된다.

이상의 연구 결과로 미루어 볼 때, 댕댕이나무 잎과 가지 추추출물은 대장암 세포주 HCT116과 SW480세포의 생육을 억제 하였으나 열매추출물은 억제활성이 나타나지 않았다. 잎과 가지 추출물은 cell migration과 wound healing assay를 통해 비정상적인 세포증식 억제를 확인하였으며, β-catenin과 TCF4 의 단백질 수준을 감소시켜 비정상적인 Wnt 신호전달을 억제를 통해 대장암세포의 생육을 억제하는 것으로 판단된다. 따라서 댕댕이나무 잎과 가지는 항암을 위한 대체보완소재 및 천연 항암제 개발을 위한 소재로 활용이 가능할 것으로 판단된다.

In this study, we evaluated the effect of branch (STB) and leave (STL) extracts from Sageretia thea on β-catenin level in human colorecal cancer cells, SW480 and lung cancer cells, A549. STB and STL dose-dependently suppressed the growth of SW480 and A549 cells. STB and STL decreased β-catenin level in both protein and mRNA level. MG132 decreased the downregulation of β-catenin protein level induced by STB and STL. However, the inhibition of GSK3β by LiCl or ROS scavenging by NAC did not block the reduction of β-catenin protein by STB and STL. Our results suggested that STB and STL may downregulate β-catenin protein level independent on GSK3β and ROS. Based on these findings, STB and STL may be a potential candidate for the development of chemopreventive or therapeutic agents for human colorectal cancer and lung cancer.

Potassium cyanate는 무기화합물로 단백질의 번역 후 과정에서 카바밀화(carbamylation)을 유도할 수 있고 이러한 카바밀화 반응은 다양한 질병 및 조건에서 세포의 사멸과 관련이 있다. 이전 연구결과에서 KCN은 사람 대장암 세포주인 HCT 116세포의 방사선 감수성을 향상시키는 것을 확인하였지만 그 기전을 명확히 규명하기에는 많이 부족한 실정이다. 본 연구에서는 방사선에 다소 저항성을 가지는 대장암 세포에서 KCN이 방사선 감수성을 향상시키고 세포사멸 시키는 기전을 확인하기 위해 2 mM의 KCN 처리 후 저 선량의 광자선을 조사하여 세포주기, 세포 생존율, 세포 사멸 관련 단백질(caspase-1, PARP) 발현량, TNF-α 분비 및 TNF-α 관련 전사인자(NF-κB)의 연관성을 확인하였다. 그 결과 KCN 처리 후 광자선을 조사한 세포에서 cas pase-3 및 PARP의 활성이 증가하고 이는 세포주기의 정지와 세포사멸을 유도하였다. 또한 이 과정에서 DN A 전사인자인 NF-κB에 의해 세포 외로 TNF-α를 지속적으로 분비하여 세포사멸에 관여함을 확인하였다. 이러한 결과들을 토대로 KCN이 radiosensitizer로서 작용할 수 있는 가능성이 있다고 사료된다.

Patients with type II diabetes mellitus are more susceptible to colorectal cancer (CRC) incidence than non-diabetics. The anti-diabetic drug metformin is most commonly prescribed for the treatment of this disease and has recently shown antitumor effect in preclinical studies. The aberrant mutational activation in the components of RAS/RAF/MEK/ERK and PI3K/AKT/mTOR signaling pathway is very frequently observed in CRC. We previously reported that metformin inhibits the phosphorylation of ERK and BEZ235, a dual inhibitor of PI3K and mTOR, has anti-tumor activity against HCT15 CRC cells harboring mutations of KRAS and PIK3CA. Therefore, we hypothesized that simultaneous inhibition of two pathways by combining metformin with BEZ235 could be more effective in the suppression of proliferation than single agent treatment in HCT15 CRC cells. Here, we investigated the combinatory effect of metformin and BEZ235 on the cell survival in HCT15 CRC cells. Our study shows that both of the two signaling pathways can be blocked by this combinational strategy: metformin suppressed both pathways by inhibiting the phosphorylation of ERK, 4E-BP1 and S6, and BEZ235 suppressed PI3K/AKT/ mTOR pathway by reducing the phosphorylation of 4E-BP1 and S6. This combination treatment synergistically reduced cell viability. The combination index (CI) values ranged from 0.44 to 0.88, indicating synergism for the combination. These results offer a preclinical rationale for the potential therapeutic option for the treatment of CRC.

Background: Astilbe chinensis (Maxim.) Franch. Et Savat. is a plant belonging to Saxifragaceae family and contains various active ingredients including astilbin and bergenin. It has been used as a traditional Korean medicine to improve fever, pain, and cough. Recently, a number of Korean medical resources have been studied for cancer and inflammation treatment, but A. chinensis (Maxim.) Franch. Et Savat. has not yet been investigated. Consequently, this study investigated the inhibitory effect of ethanol extracts from A. chinensis (Maxim.) Franch. Et Savat. (ARE) on oxidative stress and colorectal cancer using RAW264.7 and the human colorectal cancer cell line HCT-116. Methods and Results: In total, 500 ㎍/㎖ ARE reduced cell viability by 38.96 ± 1.32%, and increased caspase-3 activity by 133.08 ± 3.41% in HCT-116 cells. Moreover, TUNEL signaling and the early apoptosis ratio (34.56 ± 1.67%) increased by 500 ㎍/㎖ ARE treatment. H2O2-induced oxidative stress and cell death were diminished by 500 ㎍/㎖ ARE treatment through decreasing ROS (reactive oxygen species). Conclusions: The inhibitory effects of ARE against human colorectal cancer cells is mediated by apoptosis and caspase-3 activation, and H2O2-induced ROS generation and cell death are decreased by ARE treatment in RAW264.7 cells. However, further study is required to explore how ARE treatment is involved in the signaling pathway to decrease ROS.