• 한국동물생명공학회지
• /
• 제37권1호
• /
• pp.62-66
• /
• 2022

To date, the development of anticancer drugs has been conducted using two-dimensional (2D) cell culture systems. However, since cancer cells in the body are generated and developed in three-dimensional (3D) microenvironments, the use of 2D anticancer drug screening can make it difficult to accurately evaluate the anticancer effects of drug candidates. Therefore, as a step towards developing a cancer cell-friendly 3D microenvironment based on a combination of vinylsulfone-functionalized polyethylene glycol (PEG-VS) with dicysteine-containing crosslinker peptides with an intervening matrix metalloproteinase (MMP)-specific cleavage site, the types of MMPs secreted from human hepatocarcinoma HepG2 cells, a representative cancer cell, were analyzed transcriptionally and translationally. MMP3 was confirmed to be the most highly expressed protease secreted by HepG2 cells. This knowledge will be important in the design of a crosslinker necessary for the construction of PEG-based hydrogels customized for the 3D culture of HepG2 cells.

• Biomolecules & Therapeutics
• /
• 제13권2호
• /
• pp.113-117
• /
• 2005

Organosulfur compounds have been shown to exert an anti-cancer activity. In an attempt to develop novel chemopreventive and anti-cancer agents for liver cancer, we synthesized allylthiopyridazine derivatives. We have previously shown that allylthiopyridazine derivatives exert inhibitory effects on proliferation, invasion and migration of SK-Hep-1 hepatocarcinoma cells in vitro. The in vivo anti-tumor effect of 3-methvl-6-allylthiopy-ridazine, named as K6, was also reported. In this study, we further investigated the preclinical anti-cancer efficacy of K6 for hepatocarcinoma using nude mice xenografted with Hep-G2 hepatocellular carcinoma cells. K6(20-100 mg/kg, orally administered everyday for 30 days) markedly decreased the tumor volume of Hep-G2 cell-transplanted nude mice as evidenced by ultrasonographic and plethysmogranhic analyses. The inhibitory effect on tumor volume was lower than that exerted by doxorubicin (2 mg/kg), intravenously injected) which was used as a positive control. This study shows that K6 efficiently suppresses xenograft tumor growth, revealing K6 as apotential anti-cancer agent for suppressing in vivo progression of liver cancer. Given that hepatocarcinoma is among the most prevalent and lethal malignancies and there is no effective treatment to date, our study may contribute to the potential drug development for liver cancer.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권15호
• /
• pp.6417-6421
• /
• 2015

Viscum album var (VAV) also known as mistletoe, has long been categorized as a traditional herbal medicine in Asia. In addition to its immunomodulating activities, mistletoe has also been used in the treatment of chronic hepatic disorders in China and Korea. There are numerous reports showing that VAV possesses anti-cancer effects, however influence on human hepatocarcinoma has never been elucidated. In the present study, hot water extracts of VAV was evaluated for its potential anti-cancer effect in vitro. SK-Hep1 cells were treated with VAV (50-400ug/ml) for both 24 and 48 hours then cell viability was measured by cell counting kit-8 (CCK-8). Flow cytometry analysis was used to measure the proportion of SK-Hep1 in the different stages of cell cycle. RT-PCR and Western blot analysis were conducted to measure expression of cell cycle arrest related genes and proteins respectively. VAV dose dependently inhibited the proliferation of SK-Hep1 cells without any cytotoxicity with normal Chang liver cell (CCL-13). Flow cytometry analysis showed that VAV extract inhibited the cell cycle of SK-Hep1 cells via G1 phase arrest. RT-PCR and Western blot analysis both revealed that cyclin dependent kinase 2 (Cdk2) and cyclin D1 gene expression were significantly down regulated while p21 was upregulated dose dependently by VAV treatment. Combined down regulation of Cdk2, Cyclin D1 and up regulation of p21 can result in cell death. These results indicate that VAV showed evidence of anti-cancer activity through G1 phase cell cycle arrest in SK-Hep1 cells.

• 동의생리병리학회지
• /
• 제22권4호
• /
• pp.821-828
• /
• 2008

Onchungeum, a herbal formula, which has been used for treatment of anemia due to bleeding, discharging blood and skin disease. In the present study, it was examined the effects of extract of Onchungeum (OCE) on the growth of human hepatocarcinoma cell lines Hep3B (p53 null type) and HepG2 (p53 wild type) in order to investigate the anti-proliferative mechanism by OCE. Treatment of Hep3B and HepG2 cells to OCE resulted in the growth inhibition in a dose-dependent manner, however Hep3B cell line exhibited a relatively strong anti-proliferative activity to OEC. Flow cytometric analysis revealed that OCE treatment in Hep3B cells caused G1 phase arrest of the cell cycle, which was associated with various morphological changes in a dose-dependent fashion. RT-PCR and immunoblotting data revealed that treatment of OCE caused the down-regulation of cyclin D1 expression, however the levels of cyclin E expression were not changed by OCE. The G1 arrest of the cell cycle was also associated with the induction of Cdk inhibitor p27 by OCE. Because the p53 gene is null in Hep3B cells, it is most likely that the induction of p21 is mediated through a p53-independent pathway. Moreover, p27 detected in anti-Cdk4 and anti-Cdk2 immunoprecipitates from the OCE-treated cells, suggesting that OCE-induced p27 protein blocks Cdk kinase activities by directing binding to the cyclin/Cdk complexes. Furthermore, OCE treatment potently suppresses the phosphorylation of retinoblastoma proteins and the levels of the transcription factor E2F-1 expression. Taken together, these results indicated that the growth inhibitory effect of OCE in Hep3B hepatoma cells was associated with the induction of G1 arrest of the cell cycle through regulation of several major growth regulatory gene products.

• Molecules and Cells
• /
• 제38권6호
• /
• pp.518-527
• /
• 2015

Controlled release of medications remains the most convenient way to deliver drugs. In this study, we precipitated gold nanoparticles with quercetin. We loaded gold-quercetin into poly(DL-lactide-co-glycolide) nanoparticles (NQ) and tested the biological activity of NQ on HepG2 hepatocarcinoma cells to acquire the sustained release property. We determined by circular dichroism spectroscopy that NQ effectively caused conformational changes in DNA and modulated different proteins related to epigenetic modifications and c ell cycle control. The mitochondrial membrane potential (MMP), reactive oxygen species (ROS), cell cycle, apoptosis, DNA damage, and caspase 3 activity were analyzed by flow cytometry, and the expression profiles of different anti- and pro-apoptotic as well as epigenetic signals were studied by immunoblotting. A cytotoxicity assay indicated that NQ preferentially killed cancer cells, compared to normal cells. NQ interacted with HepG2 cell DNA and reduced histone deacetylases to control cell proliferation and arrest the cell cycle at the sub-G stage. Activities of cell cycle-related proteins, such as $p21^{WAF}$, cdk1, and pAkt, were modulated. NQ induced apoptosis in HepG2 cells by activating p53-ROS crosstalk and induces epigenetic modifications leading to inhibited proliferation and cell cycle arrest.

• 한국식품과학회지
• /
• 제43권6호
• /
• pp.787-790
• /
• 2011

본 연구는 간암세포 HepG2의 증식을 억제하고 세포 사멸을 유도하는 천연소재 발굴을 목적으로, 산겨릅나무 분획 추출물을 이용하여 간암세포 HepG2의 증식에 미치는 영향을 확인하였다. 간암세포의 생존율에 분획별 추출물이 미치는 영향을 알아보기 위하여 분획물을 24시간, 48시간 동안 농도별로 처리하여 살아있는 세포를 계수한 결과 24시간 처리 시 모든 분획에서 농도 의존적으로 유의하게 생존율이 감소하였으며 유기용매 분획물들이 수용성 용매 추출물보다 높은 생육 억제 활성을 보였다. 48시간 처리 시에도 같은 경향이 나타났으며 24시간에 비하여 세포 생존율이 비슷하거나 조금 더 감소하였다. MTT assay를 통한 세포 생존 억제율의 측정 결과 BuOH 분획물은 간암세포에 대해 농도 의존적으로 생존 억제율을 증가시켰다. 형광현미경을 이용한 세포형태 관찰에서는 BuOH 분획물의 농도 의존적으로 세포의 형태가 응축되고 무정형으로 세포사멸이 나타나 apoptotic cell의 전형적인 양상을 확인할 수 있었다. Hong 등(10)의 연구에서 산겨릅나무의 추출물 중 ethyl acetat와 butanol 분획에서 뛰어난 항산화성과 항지질과산화 활성이 있음을 밝힌 내용과 Shin 등(11)의 위암, 간암, 폐암 및 유방암 세포 등에서 암세포 생육억제효과의 보고와도 유사한 경향이 나타남을 알 수 있었다. 이상의 실험을 통해 산겨릅나무가 간암세포의 성장을 저해하는 효과가 있음을 확인할 수 있었으며 앞으로 식용 및 약용자원으로서 활용가능성이 높은 것으로 판단된다.

• BMB Reports
• /
• 제46권12호
• /
• pp.611-616
• /
• 2013

Luteolin-7-O-glucoside (LUT7G), a flavone subclass of flavonoids, has been found to increase anti-oxidant and anti-inflammatory activity, as well as cytotoxic effects. However, the mechanism of how LUT7G induces apoptosis and regulates cell cycles remains poorly understood. In this study, we examined the effects of LUT7G on the growth inhibition of tumors, cell cycle arrest, induction of ROS generation, and the involved signaling pathway in human hepatocarcinoma HepG2 cells. The proliferation of HepG2 cells was decreased by LUT7G in a dose-dependent manner. The growth inhibition was due primarily to the G2/M phase arrest and ROS generation. Moreover, the phosphorylation of JNK was increased by LUT7G. These results suggest that the anti-proliferative effect of LUT7G on HepG2 is associated with G2/M phase cell cycle arrest by JNK activation.

• KSBB Journal
• /
• 제25권6호
• /
• pp.507-512
• /
• 2010

In this study, we investigated the ability of luteolin, a plant derived flavonoid on hepatocarcinoma cell growth using HepG2 cell culture system. We found that luteolin increased the Smac/DIABLO releases, a mitochondrial protein that potentiates apoptosis. Luteolin also induced either transcriptional activity or expression of PPAR-gamma, a target of cancer growth that PPAR-gamma agonist sensitizes to apoptosis in certain cancer types. To find the possible upstream target molecules of PPAR-gamma activated by luteolin treatment, we used compound C, a specific inhibitor of AMP-activated protein kinase. Pre-treatment of Compound C significantly restored the activation or expression of PPAR-gamma stimulated by luteolin. This result indicated that AMPK signaling might be involved in the activation or expression of PPAR-gamma signaling pathway stimulated by luteolin. Moreover, we also found that luteolin inhibited the insulin-stimulated Akt phosphorylation as well as AICAR, a specific AMPK activator. These results propose that luteolin significantly induces cancer cell death through modulating survival signal pathways such as PPAR-gamma and Akt. AMPK signaling pathway may be an upstream regulator for survival signal pathways such as PPAR-gamma and Akt stimulated by luteolin.

• 생명과학회지
• /
• 제17권9호통권89호
• /
• pp.1298-1302
• /
• 2007

${\beta}-lapachone$은 남미지역에서 자생하는 Tabebuia avellanedae라는 나무의 수피에서 동정된 quinone계 물질로서 다양한 인체암세포에서 항암효과가 있는 것으로 알려져있다. 본 연구에서는 ${\beta}-lapachone$의 암 전이 억제에 대한 연구의 일환으로 HepG2 및 Hep3B 인체 간암 세포의 전이관련 유전자의 발현에 미치는 ${\beta}-lapachone$의 영향을 조사하였다. MTT assay 및 세포형태변화 관찰 결과에서 ${\beta}-lapachone$ 처리에 따라 HepG2와 Hep3B 세포들은 ${\beta}-lapachone$ 농도 의존적으로 세포의 증식이 억제되었으며 그 형태적 변형도 동반하였다. ${\beta}-lapachone$처리에 의한 암 전이 지표가 되는 IGF-lR, Tjs (ZO-1, claudin-3,-4) 및 Tj 조절인자(${\beta}-catenin$)의 발현을 RT-PCR과 Western blot analysis를 통하여 확인한 결과 ${\beta}-lapachone$ 처리가 IGF-1R의 발현 억제와 Tj 유전자 발현의 증가를 유도함으로써 ${\beta}-lapachone$이 Tj를 강화하여 암세포의 전이 억제작용을 하는 것으로 관찰 되었다. 이상의 결과는 인체 간암세포에서 ${\beta}-lapachone$의 항전이 작용의 이해에 중요한 기초 자료가 될 것으로 생각한다.

• 한국식품영양과학회지
• /
• 제37권6호
• /
• pp.714-720
• /
• 2008

본 연구에서는 동충하초(C. militaris)의 항암작용 기전 해석을 위하여 Hep3B 간암세포의 apoptosis 유발에 미치는 동충하초 열수추출물(WECM)의 영향을 조사하였으며, apoptosis 조절에 중요한 몇 가지 유전자들의 발현 및 활성 변화를 조사하였다. AECM 처리에 의한 Hep3B 세포의 증식억제는 형태적 변형을 동반한 apoptosis 유도와 연관성이 있음을 DAPI 염색을 통한 apoptotic body 출현의 증가 및 flow cytometry 분석에 의한 sub-G1 기에 속하는 세포 빈도의 증가로 확인하였다. AECM 처리에 의한 apopotosis 유도에서 Bcl-2 family에 속하는 몇 가지 유전자들의 발현은 큰 변화가 없었으나, caspase-3 및 -8의 활성이 매우 높게 증가 되었으며 이는 PARP 및 ${\beta}$-catenin 단백질의 분해와 연관성이 있었다. 또한 caspase-3 선택적 저해제인 z-DEVD-fmk로 caspase-3의 활성을 인위적으로 차단시켰을 경우, AECM에 의한 apoptois 유도 현상이 유의적으로 감소되어 AECM에 의한 Hep3B 세포의 apoptosis 유발에 caspase-3이 중요한 역할을 하고 있음을 알 수 있었다. 본 연구의 결과만으로 동충하초에 의한 간암세포의 증식억제 기전을 명확하게 제시할 수는 없으나, 이상의 결과들은 동충하초의 생화학적 항암기전 해석을 이해하는데 중요한 기초자료로서 활용될 수 있을 것으로 생각된다.