• 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.

• 생명과학회지
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• 제17권6호통권86호
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• pp.778-782
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• 2007

Diphenyleneiodonium (DPI)는 NADPH oxidase 같은 flavoenzymes의 저해제로써 널리 사용되고 있다. 본 연구에서는 인간 대장암 세포주 HCT-116 (wild-type p53)와 HT-29 (p53 mutant) 및 인간 유방암 세포주인 MCF-7(wild-type p53)의 세포성장 과정에서의 DPI의 효과를 살펴보았다. DPI는 농도 및 시간 의존적으로 암세포주의성장을 막았으며 G2/M phase에서 cell cycle arrest를 일으켰다. Cell cycle arrest의 가장 높은 값은 DPI 처리후 12 시간에서 관찰할 수 있었다. 한편 DPI는 아폽토시스 그리고 cell cycle arres 에 관여하는 유전자 발현에 관여하는 p53의 표현을 크게 증가시켰으며, 이는 DPI처리 후 6시간 후 부터 관찰할 수 있었다. 그러나 NADPH oxidase의 조합을 억제하는 catechol 계인 apocynin은 p53의 발현을 유도하지 못하였다. 이것은 DPI에 의해 유도되는 p53의 발현증가는 NADPH oxidase활성의 저해와 관련되어 있지 않다는 것을 의미한다. 결론적으로 DPI는 HCT-116, HCT-15 및 MCF-7 암세포주에서 ROS에 비 의존적으로 wild-type p53 발현의 증가를 유도하며, 이 증가된 p53은 DPI에 의해 유도되는 성장 억제 및 C2/M phase에서의 cell cycle arrset과정의 조절기전에 관여한다는 것을 시사한다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1947-1959
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• 2016

Background: Colorectal cancer (CRC) is a major cause of morbidity and mortality, being the second most common type of cancer worldwide in both men and women. It accounts yearly for approximately 9% of all new cases of cancers. Furthermore, the current chemotherapeutic regimens seem unsatisfactory, so that exploration of novel therapeutic modalities is needed. The present study was undertaken to investigate the inhibitory effects of a crude alkaloid extract (CAERS) of a medicinal herb, Rhazya stricta, on proliferation of CRC HCT116 cells and to elucidate mechanisms of action. To achieve these aims, we utilized MTT, comet, DNA laddering and gene reporter assays, along with Western blot and RT-PCR analyses. Results: We found that CAERS inhibited cell proliferation and induced apoptotic cell death in HCT116 cells. Hallmarks of morphological and biochemical signs of apoptosis were clearly evident. CAERS down-regulated DNA-binding and transcriptional activities of NF-${\kappa}B$ and AP-1 proteins, while up-regulating expression of the Nrf-2 protein. It also down-regulated expression levels of the ERK MAPK, Bcl-2, cyclin D1, CDK-4, survivin and VEGF and up-regulated levels of Bax, caspase-3/7 and -9, p53, p21, Nrf-2. Markedly, it promoted mRNA expression levels of cytoprotective genes including the hemeoxygenase-1, NAD(P)H quinine oxidoreductase 1 and UDP-glucuronyltransferase. Conclusions: These findings indicate that CAERS exerts antiproliferative action on CRC cells through induction of apoptotic mechanisms, and suggest CAERS could be a promising agent for studying and developing novel chemotherapeutic agents aimed at novel molecular targets for the treatment of CRC.

• 한국식품과학회지
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• 제45권6호
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• pp.770-776
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• 2013

본 연구에서는 호박잎 추출물의 총 폴리페놀 함량, 총 플라보노이드 함량, radical 소거활성 등과 같은 항산화성과, 암세포 증식억제 활성, 사멸유도 활성, 부착억제 활성 등과 같은 항암성을 in vitro 수준에서 검증하였다. 호박잎의 총 폴리페놀 함량은 263.4 mg gallic acid equivalent/100 g, 총 플라보노이드 함량은 73.6 mg quercetin equivalent/100 g으로 측정되었다. 호박잎 추출물은 300 ${\mu}g/mL$ 농도에서 69.4%의 ABTS radical 소거 활성이 있었는데, 이는 호박잎 추출물과 같은 농도에서의 L-ascorbic acid 활성(94.5%)의 73.4%에 해당되는 수준이었다. 한편 호박잎 추출물은 37.5, 75, 150 ${\mu}g/mL$ 농도에서 HCT116 대장암 세포와 H1299 폐암 세포의 증식을 60.6-87.9% 억제하는 농도 의존적 활성이 있었으며 이는 부분적으로 apoptosis를 유도하기 때문인 것으로 생각된다. 또한 호박잎 추출물은 150 ${\mu}g/mL$ 농도에서 HCT116 세포와 H1299 세포의 부착을 16.8-28.4% 억제하는 활성이 있었다. 이상의 결과를 통하여 호박잎 추출물은 in vitro 수준에서 항산화성 및 항암성을 가지는 것으로 생각되며, 이러한 연구 결과는 향후 관련 작용기전을 규명하기 위한 기초 자료로 이용될 수 있을 것으로 기대된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권6호
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• pp.3975-3978
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• 2013

Background: Proteases play a regulatory role in a variety of pathologies including cancer, pancreatitis, thromboembolic disorders, viral infections and many others. One of the possible strategies to combat these pathologies seems to be the use of protease inhibitors. LC-pi I, II, III and IV (Lavatera cashmerian-protease inhibitors) have been found in vitro to strongly inhibit trypsin, chymotrypsin and elastase, proteases contributing to tumour invasion and metastasis, indicated possible anticancer effects. The purpose of this study was to check in vitro anticancer activity of these four inhibitors on human lung cancer cell lines. Material and Methods: In order to assess whether these inhibitors induced in vitro cytoxicity, SRB assay was conducted with THP-1 (leukemia), NCIH322 (lung) and Colo205, HCT-116 (colon) lines. Results: LC-pi I significantly inhibited the cell proliferation of all cells tested and also LC-pi II was active in all except HCT-116. Inhibition of cell growth by LC-pi III and IV was negligible. $IC_{50}$ values of LC-pi I and II for NCIH322, were less compared to other cell lines suggesting that lung cancer cells are more inhibited. Conclusion: These investigations might point to future preventive as well as curative solutions using plant protease inhibitors for various cancers, especially in the lung, hence warranting their further investigation.

• The Korean Journal of Physiology and Pharmacology
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• 제15권1호
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• pp.1-7
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• 2011

Many anticancer agents as well as ionizing radiation have been shown to induce autophagy which is originally described as a protein recycling process and recently reported to play a crucial role in various disorders. In HCT116 human colon cancer cells, we found that curcumin, a polyphenolic phytochemical extracted from the plant Curcuma longa, markedly induced the conversion of microtubule-associated protein 1 light chain 3 (LC3)-I to LC3-II and degradation of sequestome-1 (SQSTM1) which is a marker of autophagosome degradation. Moreover, we found that curcumin caused GFP-LC3 formation puncta, a marker of autophagosome, and decrease of GFP-LC3 and SQSTM1 protein level in GFP-LC3 expressing HCT116 cells. It was further confirmed that treatment of cells with hydrogen peroxide induced increase of LC3 conversion and decrease of GFP-LC3 and SQSTM1 levels, but these changes by curcumin were almost completely blocked in the presence of antioxidant, N-acetylcystein (NAC), indicating that curcumin leads to reactive oxygen species (ROS) production, which results in autophagosome development and autolysosomal degradation. In parallel with NAC, SQSTM1 degradation was also diminished by bafilomycin A, a potent inhibitor of autophagosome-lysosome fusion, and cell viability assay was further confirmed that cucurmin-induced cell death was partially blocked by bafilomycin A as well as NAC. We also observed that NAC abolished curcumin-induced activation of extracelluar signal-regulated kinases (ERK) 112 and p38 mitogen-activated protein kinases (MAPK), but not Jun N-terminal kinase (JNK). However, the activation of ERK1/2 and p38 MAPK seemed to have no effect on the curcumin-induced autophagy, since both the conversion of LC3 protein and SQSTM1 degradation by curcumin was not changed in the presence of NAC. Taken together, our data suggest that curcumin induced ROS production, which resulted in autophagic activation and concomitant cell death in HCT116 human colon cancer cell. However, ROS-dependent activation of ERK1/2 and p38 MAPK, but not JNK, might not be involved in the curcumin-induced autophagy.

• 한국식품영양학회지
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• 제31권6호
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• pp.836-843
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• 2018

The objective of this study was to determine the effect of high hydrostatic pressure (HHP) treatment on proliferation of human cancer cell lines (MCF-7, HCT-116, PC-3 and AGS) of crude soyasaponin extracts in germinated black soybean. Black soybean was germinated and subjected to HHP, followed by preparation of crude soyasaponin extracts. Cell treatments done with extracts less than $400{\mu}g/mL$ concentrations had no significant effect on 3T3-L1 adipocyte cell viability. The inhibitory effect of crude soyasaponin extracts with germination periods and applied pressure on breast cancer cell (MCF-7), human colon cancer cell (HCT-116), human gastriccancer cell (AGS) and prostate cancer cell (PC-3) growth were investigated using MTT assay. The highest anti-proliferation of human cancer cell line of crude soyasaponin extracts was observed at 150 MPa treatment after germination for 4 days (150 MPa-Day 4). The cell viability on MCF-7, HCT-116, PC-3 and AGS cell lines of crude soyasaponin extract in 150 MPa-Day4 was 48.82%, 57.37%, 39.89% and 23.94% at $400{\mu}g/mL$, respectively. These results suggest that soyasaponin extracts from black soybean subjected to HHP after germination may mediate physiological activity.

• 한국식품과학회지
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• 제43권5호
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• pp.641-647
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• 2011

본 연구에서는 녹차의 주요 생리활성 성분인 PPE와 일반 의약품 성분 AAP, Asp 및 Ibu와의 혼용 시 일어날 수 있는 상호 작용에 의한 세포독성 변화를 장관계 정상세포와 암세포 모델에서 비교하였다. PPE는 정상 장관계 세포 INT 407 및 대장암 세포 HCT 116에 농도의존적인 독성을 나타내었고, $IC_{50}$ 수치는 각각 29.9과 57.4 ${\mu}g/mL$로써 암세포에 더욱 강력한 독성을 보였다. PPE와 200 ${\mu}M$ 이하의 저농도 약물을 HCT 116 세포에 24, 48시간 복합 투여한 결과 전체적으로 현저한 독성의 변화현상은 발생하지 않았다. HCT 116 세포에서 항산화효소 SOD 존재 시에 PPE 독성의 유의적인 감소가 관찰되었으나, 이 조건에서도 약물에 의한 유의적인 변화는 나타나지 않았다. PPE로부터 생성되는 $H_2O_2$의 양은 5 mM 이상의 Asp 또는 Ibu에 의하여 감소하였고, AAP에는 영향을 받지 않았다. INT 407 세포와 HCT 116 세포에서 mM 이상의 고농도 약물과 PPE를 복합 처리하여 세포독성변화를 본 결과 현저한 독성의 변화가 관찰되지 않았으나, INT 407 세포에서 PPE 존재 시 AAP의 독성이 다소 증가하였다. 한편 PPE와 약물을 각각 순서를 달리하여 전후로 처리하였을 때 HCT 116 세포에서의 독성이 단독처리 시 각각의 합보다 20% 이내에서 증가하였다. PPE와 빈번히 복용되는 일반 의약품 AAP, Asp 및 Ibu를 여러 조합에 의해 장관계 세포에 처리하였을 때 이들의 상호작용에 의해 일부 독성의 강화현상이 나타났으나 전체적으로 두드러진 독성발현 빛 활성변화는 발견되지 않았다.

• Journal of Microbiology and Biotechnology
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• 제31권12호
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• pp.1615-1623
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• 2021

Picropodophyllotoxin (PPT), an epimer of podophyllotoxin, is derived from the roots of Podophyllum hexandrum and exerts various biological effects, including anti-proliferation activity. However, the effect of PPT on colorectal cancer cells and the associated cellular mechanisms have not been studied. In the present study, we explored the anticancer activity of PPT and its underlying mechanisms in HCT116 cells. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was used to monitor cell viability. Flow cytometry was used to evaluate cell cycle distribution, the induction of apoptosis, the level of reactive oxygen species (ROS), assess the mitochondrial membrane potential (Δψm), and multi-caspase activity. Western blot assays were performed to detect the expression of cell cycle regulatory proteins, apoptosis-related proteins, and p38 MAPK (mitogen-activated protein kinase). We found that PPT induced apoptosis, cell cycle arrest at the G1 phase, and ROS in the HCT116 cell line. In addition, PPT enhanced the phosphorylation of p38 MAPK, which regulates apoptosis and PPT-induced apoptosis. The phosphorylation of p38 MAPK was inhibited by an antioxidant agent (N-acetyl-L-cysteine, NAC) and a p38 inhibitor (SB203580). PPT induced depolarization of the mitochondrial inner membrane and caspase-dependent apoptosis, which was attenuated by exposure to Z-VAD-FMK. Overall, these data indicate that PPT induced G1 arrest and apoptosis via ROS generation and activation of the p38 MAPK signaling pathway.

• 생명과학회지
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• 제18권8호
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• pp.1140-1146
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• 2008

본 연구에서는 항균작용, 항암작용, 항산화작용이 있는 것으로 알려져 있는 마늘을 이용하여 소비자의 요구에 부합하는 기능성 전통 고추장 개발을 목표로 마늘죽을 첨가한 고추장을 제조한 후 이에 대한 성분 분석, 항산화능력 및 항암 능력을 살펴본 결과는 다음과 같다. 고추장의 품질 기준으로 평가되는 아미노태 질소함량분석 결과, 마늘죽 첨가 고추장이 시판 고추장에 비해 높은 것으로 나타났으며 숙련된 패널을 대상으로 관능 평가한 결과 생마늘 첨가 고추장이 색을 제외한 항목에 있어 마늘 특유의 향에 대한 거리감으로 인해 낮게 평가된 반면 마늘죽 형태로 첨가한 고추장의 경우 평가항목 모두에서 높은 점수를 얻었다. 질량 분석기를 이용하여 마늘죽 첨가 고추장을 정성 분석한 결과 diallyl disulfide (C6H10S2)와 diallyl trisulfide (C6H10S3)와 같은 유효 기능성 성분인 설파이드계 화합물이 존재하는 곳으로 확인되었다. 항산화 능력은 마늘죽 첨가고추장이 라디칼 소거능 및 환원력에 있어서 모두 시판 고추장에 비해 높은 것으로 조사되었으며 특히 MTT assay법으로 위암(MKN45), 대장암(HCT116), 폐암세포(NCI-H460)를 대상으로 항암효과를 조사한 결과 마늘죽 첨가 고추장의 경우 모든 암세포에 대해 항암효과가 있는 것으로 나타났으며 특히 위암(MKN45)세포에 대한 항암효과가 가장 높은 것으로 나타났다. 결론적으로, 마늘죽 첨가 고추장(마늘 함량 23%, w/w)이 시판 고추장 및 생마늘 첨가 고추장(마늘 함량 10%, w/w)보다 항산화 및 항암효과가 높은 이유는 마늘 함유량이 높기 때문으로 생각된다.