• Molecules and Cells
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• 제28권5호
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• pp.489-494
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• 2009

We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

• 한국방사선학회논문지
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• 제13권1호
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• pp.125-132
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• 2019

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, $TNF-{\alpha}$ 분비 및 $TNF-{\alpha}$ 관련 전사인자($NF-{\kappa}B$)의 연관성을 확인하였다. 그 결과 KCN 처리 후 광자선을 조사한 세포에서 caspase-3 및 PARP의 활성이 증가하고 이는 세포주기의 정지와 세포사멸을 유도하였다. 또한 이 과정에서 DNA 전사인자인 $NF-{\kappa}B$에 의해 세포 외로 $TNF-{\alpha}$를 지속적으로 분비하여 세포사멸에 관여함을 확인하였다. 이러한 결과들을 토대로 KCN이 radiosensitizer로서 작용할 수 있는 가능성이 있다고 사료된다.

• Biomolecules & Therapeutics
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• 제31권1호
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• pp.73-81
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• 2023

Sirtuins (SIRTs) belong to the nicotinamide adenine dinucleotide (NAD+)-dependent class III histone deacetylase family. They are key regulators of cellular and physiological processes, such as cell survival, senescence, differentiation, DNA damage and stress response, cellular metabolism, and aging. SIRTs also influence carcinogenesis, making them potential targets for anticancer therapeutic strategies. In this study, we investigated the anticancer properties and underlying molecular mechanisms of a novel SIRT1 inhibitor, MHY2251, in human colorectal cancer (CRC) cells. MHY2251 reduced the viability of various human CRC cell lines, especially those with wild-type TP53. MHY2251 inhibited SIRT1 activity and SIRT1/2 protein expression, while promoting p53 acetylation, which is a target of SIRT1 in HCT116 cells. MHY2251 treatment triggered apoptosis in HCT116 cells. It increased the percentage of late apoptotic cells and the sub-G1 fraction (as detected by flow cytometric analysis) and induced DNA fragmentation. In addition, MHY2251 upregulated the expression of FasL and Fas, altered the ratio of Bax/Bcl-2, downregulated the levels of pro-caspase-8, -9, and -3 proteins, and induced subsequent poly(ADP-ribose) polymerase cleavage. The induction of apoptosis by MHY2251 was related to the activation of the caspase cascade, which was significantly attenuated by pre-treatment with Z-VAD-FMK, a pan-caspase inhibitor. Furthermore, MHY2251 stimulated the phosphorylation of c-Jun N-terminal kinase (JNK), and MHY2251-triggered apoptosis was blocked by pre-treatment with SP600125, a JNK inhibitor. This finding indicated the specific involvement of JNK in MHY2251-induced apoptosis. MHY2251 shows considerable potential as a therapeutic agent for targeting human CRC via the inhibition of SIRT1 and activation of JNK/p53 pathway.

• 한방비만학회지
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• 제12권1호
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• pp.33-47
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• 2012

Objectives This study was designed to investigate the effects of fermented soybean upon anti-inflammation, cytotoxicity, antioxidant and intestinal mucous membrane permeability by measuring the cell viability, NO (nitric oxide) production, DPPH, Polyphenol, HRP and TEER in cells like Raw 264.7 and HCT 116 using fermented soybean. Methods Raw 264.7 cell and HCT 166 cell were used in this study. And fermented soybean powders were used for the experimental group and soybean powders for the control group. There was inflammation response upon using lipopolysaccharide(LPS). Fermented soybean powders and soybean powders were in a respectively different dose added to the cells with LPS. MTT assay, NO, DPPH and Polyphenol measurement, TEER, HRP were conducted for each cell. The results of this study were presented in mean and standard deviation. Results 1. In Raw 254.7 cells added with $100{\mu}l/ml$ unfermented soybean powders, 104.95% higher than 62.59% was measured. In Raw 254.7 cells added with $100{\mu}l/ml$ fermented soybean powders, there was 74.90% measured higher than 62.59%, which was a significant result. 2. By a gradual increase of unfermented soybean powders like $0.1{\mu}l/ml$, $1.0{\mu}l/ml$, $10{\mu}l/ml$, $100{\mu}l/ml$, the measured NO were also gradually decreased $53.12{\mu}M$, $47.57{\mu}M$, $37.02{\mu}M$, $28.16{\mu}M$. In case of cells added with fermented soybean powders, $43.95{\mu}M$ NO was measured in $0.1{\mu}l/ml$ which is significant, and in other cases, mostly measured over$ 56.72{\mu}M$. 3. It was inferred that fermented soybean powders have anti-inflammatory effects of maintaining intestinal mucous membrane permeability because the measured values of cells in both groups were all higher than $133.62{\Omega}$ measured of cells added with only LPS. And measured values of cells in both groups were all lower than 2.26 measured of cells added with only LPS. 4. In case of experiment DPPH and polyphenol measurement, fermented group was all higher than unfermented group. Conclusion From the results of conducting MTT assay, NO measurement, and TEER, HRP by using cells Raw 264.7 and HCT-116, even though there was no significance in the correlation between cytotoxicity, anti-inflammatory effects, both unfermented soybean powders and fermented soybean powders were shown to have intestinal mucous membrane permeability improvement effects. This effects could be applicable for autoimmune diseases, chronic inflammatory diseases and so additional studies are expected in the future. From the results of conducting DPPH, Polyphenol measurement, Fermented soybean may be useful as potential antioxidant.

• Journal of Ginseng Research
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• 제42권3호
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• pp.288-297
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• 2018

Background: The incidence of colorectal cancer (CRC) is increasing, with metastasis of newly diagnosed CRC reported in a large proportion of patients. However, the effect of Korean Red Ginseng extracts (KRGE) on epithelial to mesenchymal transition (EMT) in CRC is unknown. Therefore, we examined the mechanisms by which KRGE regulates EMT of CRC in hypoxic conditions. Methods: Human CRC cell lines HT29 and HCT116 were incubated under hypoxic (1% oxygen) and normoxic (21% oxygen) conditions. Western blot analysis and real-time PCR were used to evaluate the expression of EMT markers in the presence of KRGE. Furthermore, we performed scratched wound healing, transwell migration, and invasion assays to monitor whether KRGE affects migratory and invasive abilities of CRC cells under hypoxic conditions. Results: KRGE-treated HT29 and HCT116 cells displayed attenuated vascular endothelial growth factor (VEGF) mRNA levels and hypoxia-inducible $factor-1{\alpha}$ ($HIF-1{\alpha}$) protein expression under hypoxic conditions. KRGE repressed Snail, Slug, and Twist mRNA expression and integrin ${\alpha}V{\beta}6$ protein levels. Furthermore, hypoxia-repressed E-cadherin was restored in KRGE-treated cells; KRGE blocked the invasion and migration of colon cancer cells by repressing $NF-{\kappa}B$ and ERK1/2 pathways in hypoxia. Conclusions: KRGE inhibits hypoxia-induced EMT by repressing $NF-{\kappa}B$ and ERK1/2 pathways in colon cancer cells.

• 대한의생명과학회지
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• 제28권3호
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• pp.200-205
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• 2022

Established cancer cell lines are widely used for developing biomarkers for the patient-specific treatment of colorectal cancer and predicting prognoses. However, cancer cell lines may exhibit different drug responses depending upon the characteristics of the cell line. Therefore, it is necessary to select a tumor cell line suitable for the purpose of the study by considering the cell characteristics. This study investigated the levels of CXCL10, which were recently been reported to play an important role in the outcome of tumor treatment, secreted by colon cancer cells. 2 × 10⁵ cells/mL of each colorectal cancer cell was seeded into a 35 mm cell culture dish. After 24 h incubation, culture supernatant was used to determine the secreted CXCL10 levels. Among six colorectal cancer cell lines (HT-29, HCT116, CaCo-2, SW620, SW480, and CT26), Caco-2 cells showed the highest level of CXCL10 secretion. HT-29 cells showed the second-highest level of CXCL10 secretion. No significantly measurable level of CXCL10 secretion was detected in HCT116 cells. These results will be helpful in investigating the molecular basis of colorectal cancer.

• 한국식품영양과학회지
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• 제45권1호
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• pp.44-51
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• 2016

본 연구에서는 패랭이꽃의 항산화 성분 함량을 측정하고 패랭이꽃 에탄올 추출물의 항산화, 항염, 항암 활성을 in vitro 수준에서 평가하고자 하였다. 패랭이꽃의 총 폴리페놀, 총 플라보노이드, 총 카로티노이드 함량은 각각 19.0 mg GAE/g, 65.7 mg QE/g, $95.0{\mu}g/g$으로 측정되었다. 패랭이꽃 추출물($1,000{\mu}g/mL$)의 DPPH radical 소거 활성은 44.1%, 철환원력은 51.1%로 같은 농도의 ascorbic acid의 활성보다는 낮았지만 의미 있는 수준의 활성을 나타내었다. 패랭이꽃 추출물은 RAW 264.7 대식세포의 NO 생성을 대조구 대비 7~23% 수준으로 억제하는 농도 의존적 활성을 나타내었고, H1299 폐암세포와 HCT116 대장암세포의 성장을 대조구 대비 각각 2~81%(48~96시간 처리 시점)와 10~80%(72시간 처리시점)로 억제하는 농도 의존적 활성 또한 나타내었다. 패랭이꽃 추출물은 암세포의 부착을 억제하는 활성이 H1299와 HCT116 세포에서 모두 나타났으나 HCT116 세포에서 나타난 활성($250{\sim}1,000{\mu}g/mL$ 이상의 농도 처리시 대조구 대비 26~40% 부착 수준)이 H1299 세포에서 나타난 활성($1,000{\mu}g/mL$ 농도 처리 시 대조구 대비 55% 부착 수준)보다 컸다. 이상의 연구 결과를 통하여 패랭이꽃 추출물은 항산화 성분 함량 및 활성이 유의미한 수준이고 세포 수준의 항염 및 항암 활성을 가지는 것으로 생각된다. 앞으로 이와 같은 연구 결과가 in vivo 수준에서 재현되는지 여부를 검증하고 관련 기전을 탐색하는 심도 있는 연구가 필요할 것으로 생각된다.

• 한국식품영양과학회지
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• 제39권1호
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• pp.36-41
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• 2010

상황버섯, 영지버섯 및 노루궁뎅이버섯 균사체로 발효된 인삼추출물이 암세포증식에 미치는 영향을 살펴보기 위하여 위암세포(MKN-45), 대장암세포(HCT116), 유방암세포(MCF-7), 폐암세포(NCIH460), 전립선암세포(PC-3) 및 간암세포(HepG2)에 농도별(0.25～1.5 mg/mL)로 처리하여 암세포 성장억제율을 측정하였다. 전립선암 세포에서의 암세포 증식억제 효과는 영지버섯 균사체발효 인삼추출물이 1.5 mg/mL 농도에서 3.07%로 가장 낮은 생존율을 나타내었으며, 상황버섯 균사체발효 인삼추출물이 35.05%, 노루궁뎅이버섯 균사체발효 인삼추출물이 44.29%의 생존율을 보였다. 폐암세포에 대한 세 가지 버섯균사체발효 인삼추출물의 1.5 mg/mL 농도에서 영지버섯 균사체발효 인삼추출물은 5.31%로 우수한 항암활성을 나타낸 반면, 상황버섯 균사체발효 인삼추출물이 53.52%, 노루궁뎅이버섯 균사체발효 인삼추출물이 35.27%의 생존율을 나타내었다. 이러한 결과로부터 영지버섯 균사체로 발효시킨 인삼추출물이 다른 균사체 인삼발효물보다 다양한 암세포에 대한 성장억제 효과가 우수함을 확인할 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권2호
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• pp.743-748
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• 2016

Background: $K^-Ras$ activation is an early event in colorectal carcinogenesis and associated mutations have been reported in about 40% of colorectal cancer patients. These mutations have always been responsible for enhancing malignancy and silencing them is associated with attenuation of tumorigenicity. Among downstream effectors are the RAF/MEK/ERK and the PI3K/Akt signaling pathways. PI3K/Akt signaling leads to reduction of apoptosis, stimulated cell growth and enhanced proliferation. Ellagic acid (EA), a naturally occurring antioxidant, has recently emerged as a promising anti-cancer agent. Purpose: To evaluate the impact of cellular genetic makeup of two colon cancer cell lines with different genetic backgrounds, HCT-116 ($K^-Ras^-/p53^+$) and Caco-2 ($K^-Ras^+/p53^-$), on response to potential anti-tumour effects of EA. In addition, the influence of $K^-Ras$ silencing in HCT-116 cells was investigated. Materials and Methods: Cellular proliferation, morphology and cell cycle analysis were carried out in addition to Western blotting for detecting total Akt and p-Akt (at Thr308 and Ser473) in the presence and absence of different concentrations of EA. Cell proliferation was also assessed in cells transfected with different concentrations of $K^-Ras$ siRNA or incubated with ellagic acid following transfection. Results: The results of the present study revealed that EA exerts anti-proliferative and dose-dependent pro-apoptotic effects. Cytostatic and cytotoxic effects were also observed. p-Akt (at Thr308 and Ser473) was downregulated. Moreover, EA treatment was found to (i) reduce $K^-Ras$ protein expression; (ii) in cells transfected with siRNA and co-treated with EA, pronounced anti-proliferative effects as well as depletion of p-Akt (at Thr308) were detected. Conclusions: Cellular genetic makeup ($K^-Ras^-/p53^-$) was not likely to impose limitations on targeting EA in treatment of colon cancer. EA had a multi-disciplinary pro-apoptotic anti-proliferative approach, having inhibited Akt phosphorylation, induced cell cycle arrest and showed an anti-proliferative potential in HCT-116 cells (expressing mutant $K^-Ras$).

• 한국식품과학회지
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• 제42권2호
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• pp.217-222
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• 2010

본 연구에서는 널리 섭취되는 식이 폴리페놀 화합물인 resveratrol과 일반의약품 성분 AAP, Asp 및 Ibu와의 혼용 시 일어날 수 있는 상호작용에 의한 세포독성 변화를 조사하였다. Resveratrol은 장관계 HCT 116 세포와 간 HepG2 세포에 농도의존적인 세포활성 감소를 초래하였고 각각 113.8 및 $135.7\;{\mu}M$의 $IC_{50}$ 수치를 보였다. 농도별 resveratrol과 AAP, Asp 또는 Ibu를 각 세포에 24시간 복합투여하였을 때 일부 유의적인 resvertrol 독성의 감소나 증가가 나타났지만 전체적으로 10% 이내의 미미한 변화를 보였다. AAP, Asp 및 Ibu의 고농도 처리시 발생하는 세포독성에 대한 resveratrol의 효과를 HepG2 세포와 IEC-6 정상장관계 세포에서 비교하였을 때 현저한 약물 독성의 변화 또한 관찰되지 않았다. HepG2 세포에 저농도의 resveratrol을 48, 72시간 처리하였을 때 유의적인 세포증식 촉진효과가 나타났으나, AAP와 복합투여시 그 효과는 소멸되었다. 한편 일반의약품 성분과 resveratrol을 각각 순서를 달리하여 전후로 세포에 처리하였을 때에도 현저한 독성의 변화는 나타나지 않았다. Resveratrol과 빈번히 복용되는 일반의약품 AAP, Asp, Ibu를 여러 조합에 의해 복합처리하여 세포독성을 평가한 결과, 이들의 상호작용에 의한 두드러진 독성발현 및 활성변화는 발견되지 않았다.