Induction of Apoptosis by Linoleic Acid Is Associated with the Modulation of Bcl-2 Family and Fas/FasL System and Activation of Caspases in AGS Human Gastric Adenocarcinoma Cells

권재임,김기영,박건영,류충호,최영현 한국식품영양과학회 2008 Journal of medicinal food Vol.11 No.1

In this study, we investigated the effects of linoleic acid (LA), a polyunsaturated fatty acid found in most veg-etable oils and certain food products, on the growth of AGS human gastric adenocarcinoma cells. LA treatment resulted in abodies, chromatin condensation, and the accumulation cells in the sub-G1 phase. LA treatment induced cyclin-dependent ki-nase inhibitor p21 in a p53-independent manner; however, this compound did not affect the cell cycle distribution. Reversetranscription-polymerase chain reaction and western blot analyses showed that treating the cells with LA caused the up-reg-ulation of pro-apoptotic Bax expression and the down-regulation of anti-apoptotic Bcl-2 expression. The apoptosis of AGScells by LA was found to be associated with an elevated Fas and Fas ligand expression in a concentration-dependent manner.Furthermore, a proteolytic activation of caspases (3, 8, and 9), and degradation/cleavage of poly(ADP-ribose) polymerase andphospholipase C-.might be involved in LA-induced apoptosis of AGS cells.

Hep3B 인간 간암세포에서 caspase 의존적이며 PI3K/Akt 신호전달의 불활성화와 관련된 β-lapachone의 세포사멸 유도

권재임,최영현,황보현 한국생명과학회 2024 생명과학회지 Vol.34 No.2

β-Lapachone is a natural quinone compound originally obtained from the bark of the lapacho tree (Tabebuia vellanedae), which has been used in traditional medicine in several South and Central American countries for treating various diseases. Although β-lapachone has been reported to have potent anticancer activity in many types of cancer cells, its effect on the proliferation of hepatocellular carcinoma (HCC) cells is still unclear. Therefore, in this study, we investigated the effect of β-lapachone on the proliferation of human HCC Hep3B cells. According to our results, the decrease in cell viability of Hep3B cells caused by β-lapachone was closely related to the induction of apoptosis, which was confirmed through changes in nuclear morphology and flow cytometry. In addition, in Hep3B cells treated with β-lapachone, the expression of Bcl-2, an anti-apoptotic factor, was decreased, while the expression of Bax, an apoptosis inducer, was increased, and the activity of the caspase cascade was also increased. However, in the presence of a pan-caspase inhibitor, β-lapachone-induced apoptosis was weakened, indicating that the induction of apoptosis by β-lapachone was caspase-dependent. Moreover, β-lapachone treatment activated extracellular-regulated kinase (ERK) signaling while inhibiting activation of the phosphoinositide 3 kinase (PI3K)/Akt pathway. Furthermore, the effect of the ERK inhibitor on suppressing the induction of apoptosis by β-lapachone was minimal, and the PI3K inhibitor significantly increased β-lapachone-induced apoptosis. The findings from this study will contribute to a better understanding of the anticancer activity of β-lapachone in HCC cells. β-lapachone은 다양한 유형의 질병을 치료하기 위해 남미 및 중미 지역의 전통 의학에서 널리 사용되어 온 Tabebuia vellanedae의 껍질에서 분리된 천연 퀴논 화합물의 일종이다. β-lapachone은 여러 유형의 암세포에서 강력한 항암 활성을 갖는 것으로 보고되었지만, 간세포암종 세포의 증식에 대한 효과는 아직 불분명하다. 따라서 본 연구에서는 β-lapachone 인간 간세포암종 Hep3B 세포의 증식에 미치는 영향을 조사하였으며, 본 연구의 결과에 의하면, β-lapachone 처리에 의한 Hep3B 세포의 세포생존율 감소는 세포사멸 유도와 밀접한 관련이 있었다. 또한, β-lapachone이 처리된 Hep3B 세포에서는 항세포사멸 인자인 Bcl-2의 발현이 감소한 반면, 세포사멸 유도 인자인 Bax의 발현은 증가하였으며, 이는 caspase cascade의 활성 증가와 연관성이 있었다. 그러나 pan-caspase 억제제가 존재하는 경우 β-lapachone에 의해 유발된 세포사멸은 약화되었으며, 이는 β-lapachone에 의한 세포사멸 유도가 caspase 의존적인 현상임을 의미한다. 아울러, β-lapachone의 처리는 ERK 경로를 활성화시키면서 PI3K/Akt 경로의 활성을 억제하였으며, β-lapachone 유도 세포사멸에 ERK 억제제의 효과는 미미했지만, PI3K 억제제는 β-lapachone에 의해 유도된 세포사멸을 유의하게 증가시켰다. 비록 생체 내 동물 모델에서의 확인이 필요하지만, 본 연구의 결과는 간세포암종 세포에서 β-lapachone의 항암 활성을 이해하는 데 유용한 자료로 활용될 것이다.

인체 간암세포에서 β-lapachone 처리에 의한 Tight Junction 관련 유전자의 변화

김성옥,권재임,김기영,김남득,최영현,Kim, Sung-Ok,Kwon, Jae-Im,Kim, Gi-Young,Kim, Nam-Deuk,Choi, Yung-Hyun 한국생명과학회 2007 생명과학회지 Vol.17 No.9

${\beta}-lapachone$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. A hallmark of cancers is 'leaky' tight junctions (Tjs). TJs mediated paracellular permeability is elevated and TJs maintained cell polarity is frequently lost. Concomitantly, TJs-associated proteins including members of the claudin family of proteins are dysregulated. Recent findings indicate that these TJs changes can contribute to cancer progression. In this study, we examined the effects of ${\beta}-lapachone$, a quinone compound obtained from the bark of the lapacho tree (Tabebuia avellanedae), on the Tjs-associated regulators in human hepatocarcinoma cell lines, HepG2 and Hep3B. ${\beta}-lapachone$ treatment downregulated the levels of insulin-like growth factor 1 receptor (IGF-lR) proteins in both HepG2 and Hep3B cells. But the levels of claudin-3 and -4 proteins were increased in ${\beta}-lapachone$-treated HepG2 and Hep3B cells. And also the zonnula occludens-l (la-I) and p-catenin protein levels by ${\beta}-lapachone$ were increased in a time-dependent manner. However, claudin-3 and -4 mRNA levels were uninhibited by ${\beta}-lapachone$ in HepG2 and Hep3B. The present results suggest that the upregulation of claudin-3 and -4 protein levels by ${\beta}-lapachone$ occurs by a post-transcriptional mechanism and points to a novel mechanism by ${\beta}-lapachone$.

산두근 추출물에 의한 A549 인체 폐암세포 apoptosis 유발에 관한 연구

권혁,지규용,엄현섭,최영현,권재임 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.1

In the present study, we investigated the pathway of apoptosis by water extract of Sophora tonkinensis Radix (SDG) in A549 human lung carcinoma cells. It was found that SDG could inhibit the cell viability and induce the apoptotic cell death of A549 cells in a dose-dependent manner as measured by hemocytometer counts, DAPI staining and flow cytometry analysis. Apoptosis of A549 cells by SDG treatment was associated with a down-regulation of anti-apoptotic Bcl-2 and Bcl-XL and inhibitor of apoptosis proteins (IAPs) expression. SDG treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant degradation and/or down-regulation of poly (ADP-ribose) polymerase (PARP) and phospholipase (PLC)-γ1 protein. Additionally, DNA fragmentation by SDG treatment was connected with the activation of inhibitor of caspase-activated DNase/DNA fragmentation factor 45 (ICAD/DFF45) protein expression. Taken together, these results indicated that the inhibitory effect of SDG in A549 cells was associated with the induction of apoptotic cell death through regulation of several major growth regulatory gene products such as Bcl-2 family expression and caspase protease activity, and SDG may have therapeutic potential in human lung cancer treatment. (Cancer Prev Res 12, 58-67, 2007) Key Words: Sophora tonkinensis, A549 cells, Apoptosis, Bcl-2, Caspase

여성 우울장애 환자의 정확한 진단을 도울 수 있는 뇌파 특징을 이용한 머신러닝 기반 컴퓨터 보조 진단 시스템 개발

이지원(Ji-Won Lee),홍승용(Seung-Yong Hong),심미선(Miseon Shim),황한정(Han-Jeong Hwang) 한국정보기술학회 2021 Proceedings of KIIT Conference Vol.2021 No.11

본 연구에서는 신경생리학적인 특징을 잘 반영하는 뇌파 기반 생체지표를 이용하여 여성 주요우울장애의 진단을 도와줄 수 있는 기계학습 기반 컴퓨터 보조 진단 시스템을 개발하였다. 이를 위하여 안정상태 뇌파 기반의 서로 다른 세 개의 생체지표(주파수 스펙트럼 밀도, 위상 동기화 지수, 및 뇌-네트워크 인덱스)를 센서 및 신호원 수준에서 추출하여 특징으로 사용하였고, 분류를 위하여 서포트 벡터 머신 분류기와 leave-one-out 교차 검증을 사용하였다. 그 결과, 9개의 센서 수준의 주파수 스펙트럼 밀도 특징을 사용하였을 때 84.69%의 높은 진단 성능을 확인하였다. 앞으로 딥러닝 알고리즘을 적용하여 본 진단 시스템의 진단 성능 및 효율성을 향상시킬 예정이다. The aim of study is to develop a resting-state electroencephalography (EEG)-based computer-aided diagnosis (CAD) system for assisting accurate diagnosis of major depressive disorder (MDD) patients using machine-learning methods. To this end, we extracted three different types of features (power spectrum density, phase locking value, and network indices) in sensor- and source-level, respectively, using resting-state eyes-closed state EEG data. A support vector machine (SVM) classifier was used to differentiate 49 MDD patients and 49-matched healthy controls with leave-one-out cross-validation method. As a result, we achieved the best classification accuracy of 84.69 % using 9 sensor-level PSD features when differentiating MDD patients and healthy controls. In the future study, we will attempt to improve not only classification performances but also the efficiency the CAD system by applying deep-learning algorithms.