• The Korean Journal of Physiology and Pharmacology
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• 제21권6호
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• pp.599-607
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• 2017

Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.

• 한국전통의학지
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• 제15권1호
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• pp.106-112
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• 2006

The aim of this study was to investigate the cytotoxic effect and its mechanism on Radix Aconiti(RA) extract in lung cancer cell lines. RA extract treatment decreased the cell viability in a dose-dependent fashions in lung cancer cells including A549, H460, H23 and H157 cells. Many investigators reported that A549 and H460 cells expressed wild-type p53, but H23 and H157 cells preserved mutated p53. After treatment with RA extract in A549 and H460 cells, we measured the expression of p53 protein levels using Western blot. analysis. In both cells treated with RA extracts, p53 protein expressions were increased in a dose-dependent manner. In our experiments, RA extracts also have cytotoxic effects in H23 and H157, which have mutated p53. Treatment with RA extract decreased bcl-2 protein expressions in both cells. These results suggest that RA extracts have cytotoxic effects via p53 expression increase and bcl-2 inhibitable pathways in A549, H460 cells and H23, H157 cells, respectively.

• Biomolecules & Therapeutics
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• 제31권3호
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• pp.285-297
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• 2023

Alzheimer's disease (AD) is a neurodegenerative disease with progressive memory loss and the cognitive decline. AD is mainly caused by abnormal accumulation of misfolded amyloid β (Aβ), which leads to neurodegeneration via a number of possible mechanisms such as down-regulation of brain-derived neurotrophic factor-tropomyosin-related kinase B (BDNF-TRKB) signaling pathway. 7,8-Dihydroxyflavone (7,8-DHF), a TRKB agonist, has demonstrated potential to enhance BDNF-TRKB pathway in various neurodegenerative diseases. To expand the capacity of flavones as TRKB agonists, two natural flavones quercetin and apigenin, were evaluated. With tryptophan fluorescence quenching assay, we illustrated the direct interaction between quercetin/apigenin and TRKB extracellular domain. Employing Aβ folding reporter SH-SY5Y cells, we showed that quercetin and apigenin reduced Aβ-aggregation, oxidative stress, caspase-1 and acetylcholinesterase activities, as well as improved the neurite outgrowth. Treatments with quercetin and apigenin increased TRKB Tyr516 and Tyr817 and downstream cAMP-response-element binding protein (CREB) Ser133 to activate transcription of BDNF and BCL2 apoptosis regulator (BCL2), as well as reduced the expression of pro-apoptotic BCL2 associated X protein (BAX). Knockdown of TRKB counteracted the improvement of neurite outgrowth by quercetin and apigenin. Our results demonstrate that quercetin and apigenin are to work likely as a direct agonist on TRKB for their neuroprotective action, strengthening the therapeutic potential of quercetin and apigenin in treating AD.

• Clinical and Experimental Pediatrics
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• 제50권7호
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• pp.686-693
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• 2007

목 적 : Mycophenolate mofetil (MMF)의 활성 대사산물인 (MPA는 IMPDH의 잠재적인 반응 억제제로써 새로운 면역치료제로 사용되고 있다. 이러한 MPA는 신경계에서 흥분독성 손상 후 뇌세포를 보호하고, 미세아교세포에서는 세포사멸사(apoptosis)를 유도하지만, 저산소성 허혈성 뇌질환에서 MPA의 효과는 아직 알려지지 않아, 본 연구에서 Rice-Vannucci 모델을 이용한 신생 백서의 저산소성 허혈성 뇌 손상과 저산소 상태의 태아 백서 뇌세포 배양에서 MPA의 뇌보호 효과를 알아보고자 실험하였다. 방 법 : 생후 7일된 백서의 좌측 총 경동맥을 결찰한 후 저산소 (8% $O_2$) 상태에서 2시간 노출하여, 저산소성 허혈성 뇌 손상을 유발하고 뇌 손상 전후에 MPA(10 mg/kg)를 투여하여 대조군과 비교하였다. 또한, 재태기간 18일된 태아 백서의 대뇌피질 세포를 배양하여 1% $O_2$ 배양기에서 저산소 상태로 세포손상을 유도하여 저산소군, 손상 전후 MPA 투여군($10{\mu}g/mL$)으로 나누어 정상산소군과 비교하였다. 세포사멸사와의 관련을 알아보기 위해서 Bcl-2, Bax, caspase-3 항체로 western blotting하였고 Bcl-2, Bax, caspase-3 primer를 이용하여 real-time PCR을 하였다. 결 과 : 형태학적으로 H&E 염색상 MPA를 투여한 군에서 뇌 보호 효과를 보였다. Western blotting과 real-time PCR을 이용한 저산소성 허혈성 뇌손상 동물 모델뿐만 아니라 저산소 상태로 태아 백서 뇌세포 배양 실험에서도 MPA 투여한 경우 caspase-3의 발현과 Bax/Bcl-2의 비율이 감소함을 보였다. 결 론 : 본 연구에서 MPA가 anti-apoptosis 작용을 통하여 주산기 저산소성 허혈성 뇌 손상에 뇌보호 역할을 하는 것을 알 수 있었고 향후 신생아 저산소성 허혈성 뇌병증의 치료에 임상적 적용이 가능하리라 생각된다.

• 대한한의학회지
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• 제36권4호
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• pp.19-34
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• 2015

Objectives: This study was performed to investigate the anti-tumor effect of O. japonicus extracts on intrahepatic cholangiocarcinoma cell line SNU-1079. Methods: Cholangiocarcinoma SNU-1079 cells were treated with various concentrations of O. japonicus DW and EtOH extracts ($0-300{\mu}g/ml$) for 24, 48 or 72 h. Cell viability was evaluated through a PMS/MTS assay, and the apoptosis rate was examined through ELISA assay and flow cytometry analysis. The mRNA expression of apoptosis- and cell cycle progression-related genes (Bcl-2, Mcl-1, Bax, Survivin, Cyclin D1, and p21) was evaluated using real-time PCR, and the caspase activity was examined using immunoblot analysis. Results: O. japonicus extracts inhibited cell proliferation and increased apoptosis rate in both ELISA assay and flow cytometry analysis. O. japonicus extracts decreased Bcl-2, Mcl-1, Survivin, and Cyclin D1 mRNA expression and increased Bax mRNA level. O. japonicus extracts also increased Caspase-3 activation. Overall, O. japonicus DW extracts were more effective than EtOH extracts. Conclusions: O. japonicus inhibited cell proliferation and induced apoptosis in SNU-1079 cells via mitochondria -mediated intrinsic pathway, which leads to Caspase-3 activation. The results indicate that O. japonicus is a potential therapeutic herb with anti-tumor effect against intrahepatic cholangiocarcinoma.

• 생명과학회지
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• 제19권7호
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• pp.892-898
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• 2009

Mifepristone (MIF)은 프로게스테론 유사체이며, 강한 항프로게스테론 효과 때문에 전립선암 치료에 사용되고 있다. 본 연구에서는 MIF의 세포독성효과가 세포 내 칼슘농도 조절에 의한 것임을 밝힌다. 5-40 $\mu$M의 MIF를 처리 시 LNCaP 전립선암세포의 성장이 농도와 시간의존적으로 감소하였다. 반대로, 세포 내 칼슘의 레벨은 MIF의 처리시간과 농도도 의존적으로 증가하였다. MIF를 처리한 세포를 PI 혹은 Hoechst로 염색한 결과, 전형적인 세포자살의 징후인 응축된 염색질과 핵 조각단편들이 관찰되었다. 이들 세포자살징후 역시 MIF의 처리시간과 농도가 증가 할수록 심화되었다. 세포자살에 직접적으로 관여하는 중요한 단백질인 Bcl-2 그룹 단백질의 발현을 조사해 본 결과, 세포자살 억제단백질인 Bcl-2의 발현은 MIF처리시 치명적으로감소하였고, 대신에 세포자살 촉진단백질인 Bax의 발현은 2배로 증대되었다. 이상의 결과로 보아 MIF의 세포독성효과는 세포 내의 칼슘조절에 따른 세포자살에 의한 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권5호
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• pp.2661-2665
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• 2016

Silibinin is a natural polyphenol with high antioxidant and anticancer properties. In this study, its influence on two of the most commonly employed human breast cancer cell lines, MCF-7 and T47D, and one non-malignant MCF-10A cell line, were investigated and compared. Cell viability, the cell cycle distribution and apoptosis induction were analyzed by MTT and flow cytometry, respectively. The effect of silibinin on PTEN, Bcl-2, P21, and P27 mRNAs expression was also investigated by real-time RT-PCR. It was found that silibinin caused G1 cell cycle arrest in MCF-7 and MCF-10A cells but had no effect on the T47D cell cycle. Silibinin induced cytotoxic and apoptotic effects in T47D cells more than the MCF-7 cells and had no cytotoxic effect in MCF-10A cells under the same conditions. Silibinin upregulated PTEN in MCF-7 and caused slightly increased P21 mRNA expression in T47D cells and slightly increased PTEN and P21 expression in MCF-10A cells. Bcl-2 expression decreased in all of the examined cells under silibinin treatment. P27 mRNA expression upregulated in T47D and MCF-10A cells under silibinin treatment. PTEN mRNA in T47D and P21 and P27 mRNAsin MCF-7 were not affected by silibinin. These results suggest that silibinin has mostly different inhibitory effects in breast cancer cells and might be an effective anticancer agent for some cells linked to influence on cell cycle progression.

• Asian Pacific Journal of Cancer Prevention
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• 제17권4호
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• pp.1817-1821
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• 2016

Objectives: To observed the effects of ginsenoside -Rh2 (GS-Rh2) on proliferation and apoptosis of side population (SP) human gastric cancer SGC-7901 cells. Materials and Methods: SGC-7901 SP and Non-SP cells were sorted by flow cytometry and assessed using the cck-8 method. Expression of apoptosis-related proteins Bax and Bcl-2 of SP before and after the intervention was determined by Western-blotting. Results: It was found that the proliferation of SP was significantly faster than that of NSP (P<0.05). In addition, GS-Rh2 inhibited proliferation of gastric cancer SP cells, induced cell cycle arrest and cell apoptosis, and changed the expression of BAX/Bcl-2 proteins in a time-dependent and concentration-dependent manner (P<0.05). Conclusions: With increase of GS-Rh2 dose, GS-Rh2 gradually inhibit the proliferation of SGC-7901 SP cells, which have high proliferation rate, through G1/G0 phase arrest, followed by apoptosis which involves the up-regulation of Bax and the down-regulation of Bcl-2.

• Journal of Ginseng Research
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• 제42권2호
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• pp.133-143
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• 2018

Background: AD-2 (20(R)-dammarane-3b, 12b, 20, 25-tetrol; 25-OH-PPD) is a ginsenoside and isolated from Panax ginseng, showing anticancer activity against extensive human cancer cell lines. In this study, effects and mechanisms of 1C ((20R)-3b-O-(L-alanyl)-dammarane-12b, 20, 25-triol), a modified version of AD-2, were evaluated for its development as a novel anticancer drug. Methods: MTT assay was performed to evaluate cell cytotoxic activity. Cell cycle and levels of reactive oxygen species (ROS) were determined using flow cytometry analysis. Western blotting was employed to analyze signaling pathways. Results: 1C concentration-dependently reduces prostate cancer cell viability without affecting normal human gastric epithelial cell line-1 viability. In LNCaP prostate cancer cells, 1C triggered apoptosis via Bcl-2 family-mediated mitochondria pathway, downregulated expression of mouse double minute 2, upregulated expression of p53 and stimulated ROS production. ROS scavenger, N-acetylcysteine, can attenuate 1C-induced apoptosis. 1C also inhibited the proliferation of LNCaP cells through inhibition on $Wnt/{\beta}-catenin$ signaling pathway. Conclusion: 1C shows obvious anticancer activity based on inducing cell apoptosis by Bcl-2 family-mediated mitochondria pathway and ROS production, inhibiting $Wnt/{\beta}-catenin$ signaling pathway. These findings demonstrate that 1C may provide leads as a potential agent for cancer therapy.

• 한국응용과학기술학회지
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• 제37권3호
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• pp.418-428
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• 2020

본 연구는 알츠하이머(Alzheimer's disease: AD) 형질전환 생쥐를 대상으로 저항성 운동(resistance exercise: RE)이 해마의 베타 아밀로이드(β-amyloid: Aβ) 단백질 대사, 신경세포사멸 및 인지기능에 미치는 영향을 확인하는데 목적이 있다. AD 비 형질전환 생쥐(non-transgenic: non-tg, n=14)와 형질전환 생쥐(transgenic: Tg, n=14)를 무선 배정하여 비 형질전환 생쥐 대조군 (non-tg-control: NTC, n=7), 비 형질전환 생쥐 저항성 운동군(non-tg-RE: NTRE, n=7), 형질전환 대조군(tg-control: TC, n=7) 및 형질전환 저항성 운동군(tg-RE: TRE, n=7)으로 구분하였다. RE는 특수 제작한 사다리 저항성 운동 기구를 사용하여 점진적으로 set 수를 증가시켜 총 8주간 실시하였다. 운동 후 인지기능 능력을 평가하기 위한 수중미로검사와 Aβ 단백질 대사, 신경세포사멸 지표 및 SIRT1/PGC-1α 단백질 발현 수준을 확인하였다. 수중미로검사 결과 거리와 시간 모두 TC 집단에서 유의하게 증가 되었지만 RE를 실시한 TRE 집단에서 거리와 시간이 감소 되어 인지능력이 개선된 것으로 확인되었다. 또한, TC 집단에서 증가된 Aβ 단백질 발현은 RE를 통해 감소하는 것으로 나타났다. 신경세포사멸 관련 단백질인 Bcl-2/Bax ratio는 TC 집단에서 유의하게 감소되어 신경세포사멸이 증가 된 것으로 나타났지만 RE는 Bcl-2/Bax ratio을 증가시켜 신경세포사멸을 감소시킨 것으로 확인되었다. TC 집단에서 증가된 BACE1 및 ROCK1과 감소된 ADAM10과 RARβ 단백질 발현은 RE를 통해 감소되거나 증가 된 것으로 나타났고, SIRT1/PGC-1α 단백질 발현은 TC 집단에서 감소 되었지만 RE를 통해 증가 된 것으로 나타났다. 따라서 8주간의 RE는 AD의 병리학적 특징인 Aβ 단백질 발현을 감소시키고 관련 생성 기전들을 조절하여(SIRT1/PGC-1α 기전 활성, 아밀로이드 생성기전 억제, 비-아밀로이드 생성기전 활성) 신경세포사멸 억제시키고 결과적으로 인지기능을 개선 시킬 수 있는 효과적인 운동 방법이라고 생각된다.