• 생명과학회지
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• 제28권11호
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• pp.1268-1276
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• 2018

Cytidine analog decitabine (DEC)은 핵산 합성의 억제제로서 골수이형성 증후군 및 급성 골수성 백혈병 치료제로 사용되고 있다. 산화질소 합성에서 번역 단계를 억제하는 것으로 알려진 ammonium pyrrolidine dithiocarbamate (PDTC)는 $NF-{\kappa}B$의 대표적인 억제제이다. 본 연구에서는 인체 위암 AGS 세포를 대상으로 DEC와 PDTC의 병용 처리에 따른 세포증식 억제 기전을 조사하였다. 본 연구의 결과에 따르면 PDTC에 의한 AGS 세포의 증식 억제 효과는 DEC에 의해 농도 의존적으로 유의하게 증가하였으며, 이는 G2/M기의 세포주기 정지 및 apoptosis 유도와 관련이 있었다. PDTC와 DEC의 병용 처리에 의한 세포 사멸의 유도는 DNA 손상 유도와 관련이 있음을 H2AX의 인산화 증가로 확인하였다. 아울러 PDTC와 DEC의 병용 처리는 미토콘드리아 막 전위의 파괴를 유도하고, 세포 내 활성산소종(ROS)의 생성과 Bax의 발현을 향상시키고, Bcl-2 발현을 감소시켰으며 미토콘드리아에서 세포질로의 cytochrome c 유출을 증가시켰다. 또한 PDTC과 DEC의 병용 처리는 외인성 및 내인성 apoptosis 개시 caspase에 해당하는 caspase-8과 caspase-9의 활성뿐만 아니라 caspase-3의 활성화와 PARP 단백질의 분해를 유도하였다. 결론적으로 본 연구의 결과는 PDTC와 DEC의 병용 처리가 DNA 손상을 유발하고, ROS 증가와 연계된 외인성 및 내인성 apoptosis 사멸 경로를 활성화시킴으로써 AGS 세포의 증식을 억제하였음을 의미한다.

• 생명과학회지
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• 제34권2호
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• pp.94-104
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• 2024

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-lapa-chone의 항암 활성을 이해하는 데 유용한 자료로 활용될 것이다.

• 대한한방내과학회지
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• 제32권4호
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• pp.519-529
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• 2011

Objectives : This study was performed to investigate the effects of Injinchunggan-tang on cell growth and apoptosis in human hepatic stellate cell line LX2. Materials and Methods : Hepatic stellate cells were treated with various concentrations of Injinchunggan-tang extract for 24, 48 and 72 hours. The extraction was done with distilled water. After the treatment, cell viability, proliferation, apoptosis, caspase activity, caspase inhibitor and the mRNA of the Bcl-2, and Bax with ${\beta}$-actin were measured by using MTT assay, apoptosis assay and RT-PCR. Results : Proliferation, and mRNA expression of the hepatic stellate cells were inhibited by Injinchunggan-tang treatment in a dose-dependent manner. This indicates the prescription has inhibitory effect on fibrogenesis of the liver by regulating the fibrogenesis-associated genes in transcription. Cell viability was inhibited in time- and dose-dependent manners. Conclusions : These results suggest that Injinchunggan-tang would be beneficial in the treatment of cirrhotic patients as well as for the patients with chronic hepatitis.

• Archives of Pharmacal Research
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• 제27권11호
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• pp.1147-1153
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• 2004

Bladder cancer is a common cancer with high risk of recurrence and mortality. Intravesicle chemotherapy after trans-urethral resection is required to prevent tumor recurrence and progression. It has been known that antioxidants enhance the antitumor effect of bacillus Calmette-Guerin (BCG), the most effective intravesical bladder cancer treatment. Capsaicin, the major pungent ingredient in genus Capsicum, has recently been tried as an intravesical drug for overactive bladder and it has also been shown to induce apoptotic cell death in many cancer cells. In this study, we investigated the apoptosis-inducing effect and alterations in the cellular redox state of capsaicin in MBT-2 murine bladder tumor cells. Capsaicin induced apoptotic MBT-2 cell death in a time- and dose-dependent manner. The capsaicin-induced apoptosis was blocked by the pretreatment with Z-VAD-fmk, a broad-range caspase inhibitor, or Ac-DEVD-CHO, a caspase-3 inhibitor. In addition to the caspase-3 activation, capsaicin also induced cytochrome c release and decrease in Bcl-2 protein expression with no changes in the level of Bax. Furthermore, capsaicin at the concentration of inducing apoptosis also markedly reduced the level of reactive oxygen species and lipid peroxidation, implying that capsaicin may enhance the antitumor effect of BCG in bladder cancer treatment. These results further suggest that capsaicin may be a valuable intravesical chemotherapeutic agent for bladder cancers.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 추계학술대회
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• pp.31-31
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• 2018

Benign prostatic hyperplasia (BPH), which is the most common disorder in elderly men, involves androgenic hormone imbalance with chronic inflammation that causes imbalance between cell apoptosis and cell proliferation. As the root cause of the BPH remains unclear and synthetic drugs for treatment of BPH have undesirable side effects, the development of effective alternative medicines has been considered. Chinese Skullcap has been considered natural remedy to treat pyrexia, micturition disorder and inflammation. Although skullcap has effective properties on various diseases, the effects and molecular mechanism of Skullcap on BPH are not fully understood. Therefore, in this study, we evaluated the efficacy of Chinese Skullcap root extract (SRE) in testosterone-induced BPH rats. Compared with the untreated group, the SRE treatment group suppressed pathological alterations, such as prostate growth and increase in serum dihydrotestosterone and $5{\alpha}$-reductase levels. Furthermore, SRE significantly decreased the expression of androgen receptor and proliferating cell nuclear antigen. SRE also restored Bax/Bcl-2 balance. These effect of SRE was more prevalent than commercial $5{\alpha}$-reductase inhibitor, finasteride. Taken together, we propose that SRE suppresses abnormal androgen events in prostate tissue and inhibits the development of BPH by targeting inflammation- and apoptosis-related markers. These finding strengthens that SRE could be used as plant-based $5{\alpha}$-reductase inhibitory alternative.

• 한국식품위생안전성학회지
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• 제27권4호
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• pp.406-414
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• 2012

본 연구에서는 fucoidan이 macrophage의 활성과 항암효과를 확인하기 위하여 수행되었다. Fucoidan을 Raw 264.7 세포에 처리한 후 MTT assay로 측정한 결과 고농도 $100{\mu}g/mL$까지 세포독성은 없었으며, NO 및 TNF-${\alpha}$의 분비를 농도 유의적으로 증가시켰다. 또한 AGS 위암 세포 성장저해효과를 확인하기 위하여 MTT assay를 하였고 그 결과 농도와 시간 의존적으로 암 세포 성장이 유의적으로 감소하였다. Apoptosis로 인해 암 세포 성장이 감소하였는지 확인하기 위하여 DAPI 염색을 한 결과 apoptotic body와 세포질 응축이 시간 의존적으로 증가하는 것을 확인하였다. 또한 fucoidan은 미토콘드리아의 투과율을 향상시키며 미토콘드리아에서 방출되는 cytochrome c의 발현을 증가시켰다. Western blotting의 결과 시간 의존적으로 anti-apoptotic 분자인 Bcl-2와 XIAP 발현 감소와 반대로 pro-apoptotic 분자인 Bax 발현이 증가하였다. Cleaved-caspase-9의 발현이 증가하였으며 Akt의 인산화는 시간 의존적으로 감소하였다. Caspase 억제제인 z-VAD-FMK 처리 시 Bax, caspase-9의 발현을 감소시켜 apoptosis 유도를 억제하였으며 이러한 결과는 caspase가 apoptosis 유도에 중요한 역할을 하는 것으로 나타낸다. 본 실험의 AGS 위암 세포주에서 대조군에 비하여 fucoidan 처리군에서 면역세포 활성 및 AGS 위암 세포주에서 caspase 활성을 통해 apoptosis를 유도하는 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4519-4525
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• 2014

Oleanolic acid (OA) is a nutritional component widely distributed in various vegetables. Although it has been well recognized for decades that OA exerts certain anti-tumor activity by inducing mitochondria-dependent apoptosis, it is still unclear that what molecular signaling is responsible for this effect. In this study, we employed cancer cell lines, A549, BXPC-3, PANC-1 and U2OS to elucidate the molecular mechanisms underlying OA anti-tumor activity. We found that activation of MAPK pathways, including p-38 MAPK, JNK and ERK, was triggered by OA in both a dose and time-dependent fashion in all the tested cancer cells. Activation was accompanied by cleavage of caspases and PARP as well as cytochrome C release. SB203580 (p38 MAPK inhibitor), but not SP600125 (JNK inhibitor) and U0126 (ERK inhibitor), rescued the pro-apoptotic effect of OA on A549 and BXPC-3 cells. OA induced p38 MAPK activation promoted mitochondrial translocation of Bax and Bim, and inhibited Bcl-2 function by enhancing their phosphorylation. OA can induce reactive oxygen species (ROS)-dependent ASK1 activation, and this event was indispensable for p38 MAPK-dependent apoptosis in cancer cells. In vivo, p38 MAPK knockdown A549 tumors proved resistant to the growth-inhibitory effect of OA. Collectively, we elucidated that activation of ROS/ASK1/p38 MAPK pathways is responsible for the apoptosis stimulated by OA in cancer cells. Our finding can contribute to a better understanding of molecular mechanisms underlying the antitumor activity of nutritional components.

• 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.

• 한국발생생물학회지:발생과생식
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• 제23권1호
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• pp.43-54
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• 2019

We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipulation of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; $100{\mu}M$) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number ($63.0{\pm}7.2$) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and $41.7{\pm}3.1$, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.

• International Journal of Oral Biology
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• 제45권4호
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• pp.169-178
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• 2020

L-ascorbic acid (L-AA; vitamin C) induces apoptosis in cancer cells. This study aimed to elucidate the molecular mechanisms of L-AA-induced apoptosis in human laryngeal epidermoid carcinoma Hep-2 cells. L-AA suppressed the viability of Hep-2 cells and induced apoptosis, as shown by the cleavage and condensation of nuclear chromatin and increased number of Annexin V-positive cells. L-AA decreased Bcl-2 protein expression but upregulated Bax protein levels. In addition, cytochrome c release from the mitochondria into the cytosol and activation of caspase-9, -8, and -3 were enhanced by L-AA treatment. Furthermore, apoptosis-inducing factor (AIF) and endonuclease G (EndoG) were translocated into the nucleus during apoptosis of L-AA-treated Hep-2 cells. L-AA effectively inhibited the constitutive nuclear factor-κB (NF-κB) activation and attenuated the nuclear expression of the p65 subunit of NF-κB. Interestingly, L-AA treatment of Hep-2 cells markedly activated Akt and mitogen-activated protein kinase (MAPK; extracellular signal-regulated kinase 1/2, p38, and c-Jun N-terminal kinase [JNK]) and and LY294002 (Akt inhibitor), SB203580 (p38 inhibitor) or SP600125 (a JNK inhibitor) decreased the levels of Annexin V-positive cells. These results suggested that L-AA induces the apoptosis of Hep-2 cells via the nuclear translocation of AIF and EndoG by modulating the Bcl-2 family and MAPK/Akt signaling pathways.