• 대한화학회지
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• 제66권4호
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• pp.298-304
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• 2022

The present study was designed to investigate the molecular mechanisms of DK-5-62, a novel (-)-catechin derivative on HCT116 human colorectal cancer cells. DK-5-62 inhibited the proliferation in dose- and time-dependent manner accompanied by the morphological changes. Effects of DK-5-62 appeared to be mediated by the induction of apoptosis, as manifested through DNA-binding dye Hoechst 33258 staining. Analysis of the mechanism of these events indicated that DK-5-62-treated cells exhibited an increased ratio of Bax/Bcl-2, resulting in the activation of caspase-9, caspase-3, and poly-ADP-ribose polymerase in a dose-dependent manner. Moreover, DK-5-62-induced apoptosis was accompanied by phosphorylation of the mitogen-activated protein kinase family, c-Jun N-terminal kinase, p38, and extracellular signal-regulated kinase. These results suggest that HCT116 cells are moderately sensitive to growth inhibition by DK-5-62 via apoptosis, as evidenced by activation of ERK/p38/Bcl-2 family signaling, as well as alteration in caspase-9 and caspase-3.

• Asian Pacific Journal of Cancer Prevention
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• 제15권13호
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• pp.5201-5205
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• 2014

Emodin, a natural anthraquinone isolated from the traditional Chinese medicine Radix rhizoma Rhei, can induce apoptosis in many kinds of cancer cells. This study demonstrated that emodin induces apoptosis in human colon cancer HCT116 cells by provoking oxidative stress, which subsequently triggers a p53-mitochondrial apoptotic pathway. Emodin induced mitochondrial transmembrane potential loss, increase in Bax and decrease in Bcl-2 expression and mitochondrial translocation and release of cytochrome c to cytosol in HCT116 cells. In response to emodin-treatment, ROS increased rapidly, and subsequently p53 was overexpressed. Pretreatment with the antioxidant NAC diminished apoptosis and p53 overexpression induced by emodin. Transfecting p53 siRNA also attenuated apoptosis induced by emodin, Bax expression and mitochondrial translocation being reduced compared to treatment with emodin alone. Taken together, these results indicate that ROS is a trigger of emodin-induced apoptosis in HCT116 cells, and p53 expression increases under oxidative stress, leading to Bax-mediated mitochondrial apoptosis.

• Molecules and Cells
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• 제43권7호
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• pp.662-670
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• 2020

We have investigated the involvement of the pre-mRNA processing factor 4B (PRP4) kinase domain in mediating drug resistance. HCT116 cells were treated with curcumin, and apoptosis was assessed based on flow cytometry and the generation of reactive oxygen species (ROS). Cells were then transfected with PRP4 or pre-mRNA-processing-splicing factor 8 (PRP8), and drug resistance was analyzed both in vitro and in vivo. Furthermore, we deleted the kinase domain in PRP4 using Gateway™ technology. Curcumin induced cell death through the production of ROS and decreased the activation of survival signals, but PRP4 overexpression reversed the curcumin-induced oxidative stress and apoptosis. PRP8 failed to reverse the curcumin-induced apoptosis in the HCT116 colon cancer cell line. In xenograft mouse model experiments, curcumin effectively reduced tumour size whereas PRP4 conferred resistance to curcumin, which was evident from increasing tumour size, while PRP8 failed to regulate the curcumin action. PRP4 overexpression altered the morphology, rearranged the actin cytoskeleton, triggered epithelial-mesenchymal transition (EMT), and decreased the invasiveness of HCT116 cells. The loss of E-cadherin, a hallmark of EMT, was observed in HCT116 cells overexpressing PRP4. Moreover, we observed that the EMT-inducing potential of PRP4 was aborted after the deletion of its kinase domain. Collectively, our investigations suggest that the PRP4 kinase domain is responsible for promoting drug resistance to curcumin by inducing EMT. Further evaluation of PRP4-induced inhibition of cell death and PRP4 kinase domain interactions with various other proteins might lead to the development of novel approaches for overcoming drug resistance in patients with colon cancer.

• 생명과학회지
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• 제24권11호
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• pp.1244-1251
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• 2014

길경(桔梗, Platycodon grandiflorum)은 도라지의 뿌리로 항염증, 항알러지, 면역 반응, 당뇨, 고지혈증 및 항암 효과 등을 가지고 있는 것으로 알려져 있다. 하지만 길경의 항암 효과에 대한 연구는 미미하며, 길경이 유발하는 autophagy에 대한 연구는 되어 있지 않다. 본 연구에서는 HCT-116 대장암 세포주에서 길경 추출물이 autophagy와 apoptosis를 유발하면서 세포 성장을 억제하는지의 여부를 조사하였다. 길경 추출물은 농도 및 시간의존적으로 세포의 증식을 억제하였으며, 길경 추출물에 의해 나타나는 apoptosis는 caspase의 활성이 부분적으로 관여되어 있음을 알 수 있었다. 또한, 길경 추출물의 처리는 autophagy에 의해 나타나는 공포를 형성하면서 autophagy와 관련되어 있는 여러 단백질의 발현 조절 및 LC3 단백질의 축적이 동반되었다. 길경 추출물에 의해 유도되는 autophay와 apoptosis의 관계를 알아보기 위해서 3-MA나 bafilomycin A1을 처리하여 autophagy를 억제하였을 때 apoptosis가 유의적으로 증가됨을 알 수 있었다. 흥미롭게도 bafilomycin A1을 처리한 결과에서 길경 추출물에 의한 세포성장 억제가 뚜렷하게 회복되는 양상을 보였다. 따라서 본 연구의 결과는 HCT-116 세포에서 길경 추출물에 의해 유도된 autophagy는 세포 보호적인 작용이 아닌 autophagic cell death이며, 길경 추출물이 대장암 세포주에서 암세포의 사멸을 유도하는 효과적인 대안이 될 수 있음을 알 수 있었다.

• 한국식품위생안전성학회지
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• 제31권4호
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• pp.294-298
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• 2016

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 Coscinoderma sp.의 작용 및 그 기전을 살펴보았다. 특히, 암 억제유전자 p53의 발현을 억제시킨 세포주(HCT116 p53KO과 RKO-E6)에서의 차이점을 비교하였다. 세포생존률 시험에서 Coscinoderma sp. 추출물은 p53의 유무와 상관없이 암세포의 증식을 억제하였음을 확인하였다. 이 암세포 증식 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.

• 한국식품영양과학회지
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• 제43권1호
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• pp.86-92
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• 2014

Cordycepin은 C. militaris의 주요 생리활성 물질로서 인체 면역기능 강화, 항염증, 항산화, 항노화 및 항암활성을 포함한 다양한 약리효능이 있는 것으로 알려져 있다. 본 연구에서는 HCT116 대장암세포를 이용하여 암전이의 주요 과정인 암세포의 이동성 및 침윤성에 미치는 cordycepin의 효능에 관하여 조사하였다. 본 연구의 결과에 의하면 세포독성이 없는 범위에서 cordycepin은 HCT116 세포의 이동성과 침윤성을 유의적으로 억제하였다. RT-PCR 및 Western blotting 결과에 의하면 cordycepin은 TJs의 주요 구성인자인 claudin family 인자들의 발현을 억제하였으며, 이는 TJ의 전기적 저항성의 증대와 연관이 있었다. Cordycepin은 또한 MMP-2 및 -9의 발현과 활성을 저해함과 동시에 TIMP-1 및 -2의 발현은 증가시켰다. 따라서 cordycepin에 의한 HCT116 대장암세포의 전이능 억제는 TJ의 견고성 증대와 MMPs의 활성 억제와 연관성이 있음을 알 수 있었다.

• Natural Product Sciences
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• 제22권3호
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• pp.209-215
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• 2016

Kigelia africana (Lam.) Benth. (Bignoniaceae) is a flowering plants in South, Central and West Africa and commonly known as the sausage tree (Eng.); worsboom (Afr.); umVunguta, umFongothi (Zulu); Modukguhlu (North Sotho); Muvevha (Venda). The dried, powdered fruits are used as dressing for wounds and ulcers, haemorrhoids, rheumatism, purgative, skin-firming, lactation in breast-feeding mothers. The aim of this study is to investigate the cytotoxic and apoptotic potentials of 70% ethanolic extracts of Kigelia africana fruits in HCT116 human colon cancer cells. Treatment of Kigelia africana fruits with various concentrations resulted in a sequence of characteristic of apoptosis, including loss of cell viability and morphological changes. Flow cytometry analysis showed Kigelia africana fruits increased the sub-G1 phase (apoptosis) population. Apoptosis confirmed by annexin V-fluorescein isothiocyanate and propidium iodide double staining in HCT116 human colon cancer cell lines. Moreover, analysis of the mechanism indicated that Kigelia africana fruits showed an increased Bax and Bcl-2 expressions in a dose-dependent manner, resulting in activation of hallmarks of apoptotic events, caspase-3, caspase-9 and cleaved poly-ADP-ribose polymerase. This is the first report to demonstrate the cytotoxicity of Kigelia africana fruits on HCT116 human colon cancer cells.

• Biomolecules & Therapeutics
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• 제32권1호
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• pp.104-114
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• 2024

Licochalcone C (LCC; PubChem CID:9840805), a chalcone compound originating from the root of Glycyrrhiza inflata, has shown anticancer activity against skin cancer, esophageal squamous cell carcinoma, and oral squamous cell carcinoma. However, the therapeutic potential of LCC in treating colorectal cancer (CRC) and its underlying molecular mechanisms remain unclear. Chemotherapy for CRC is challenging because of the development of drug resistance. In this study, we examined the antiproliferative activity of LCC in human colorectal carcinoma HCT116 cells, oxaliplatin (Ox) sensitive and Ox-resistant HCT116 cells (HCT116-OxR). LCC significantly and selectively inhibited the growth of HCT116 and HCT116-OxR cells. An in vitro kinase assay showed that LCC inhibited the kinase activities of EGFR and AKT. Molecular docking simulations using AutoDock Vina indicated that LCC could be in ATP-binding pockets. Decreased phosphorylation of EGFR and AKT was observed in the LCC-treated cells. In addition, LCC induced cell cycle arrest by modulating the expression of cell cycle regulators p21, p27, cyclin B1, and cdc2. LCC treatment induced ROS generation in CRC cells, and the ROS induction was accompanied by the phosphorylation of JNK and p38 kinases. Moreover, LCC dysregulated mitochondrial membrane potential (MMP), and the disruption of MMP resulted in the release of cytochrome c into the cytoplasm and activation of caspases to execute apoptosis. Overall, LCC showed anticancer activity against both Ox-sensitive and Ox-resistant CRC cells by targeting EGFR and AKT, inducing ROS generation and disrupting MMP. Thus, LCC may be potential therapeutic agents for the treatment of Ox-resistant CRC cells.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.491-497
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• 2018

Lipopolysaccharide (LPS), a component of the cell wall of gram-negative bacteria, elicits the secretion of cytokines, such as interferons, that stimulate the host defense system. Previously, we demonstrated that interferons induce interferon regulatory factors (IRFs) 1, 3, and 7, which regulate the transcription of Noxa and alter the expression profiles of Bcl-2 family proteins in tumors. However, the immediate consequences of LPS stimulation on Noxa and BH3 expression in tumor cells remain uncharacterized. In this study, we determined that LPS induced Noxa expression in CT26 cells. Furthermore, studies in HCT116 parental and HCT116 p53-deficient cells revealed that LPS-mediated Noxa was independent of p53. Meanwhile, IRF1, 3, and 7 in CT26, HCT116 parental, and HT116 p53-deficient cells were upregulated by LPS stimulation, suggesting that LPS induces the expression of these IRFs in a p53-independent manner. The responsiveness of IRF1, 3, 4, and 7 binding to the Noxa promoter region to LPS indicated that IRF1, 3, and 7 activated Noxa expression, whereas IRF4 repressed Noxa expression. Together, these results suggest that LPS directly affects Noxa expression in tumor cells through IRFs, implicating that it may contribute to LPS-induced tumor regression.

• Nutrition Research and Practice
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• 제8권5호
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• pp.487-493
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• 2014

BACKGROUND/OBJECTIVES: D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo. MATERIALS/METHODS: The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis. RESULTS: At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-${\kappa}B$, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice. CONCLUSIONS: Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.