• The Korean Journal of Physiology and Pharmacology
• /
• 제24권6호
• /
• pp.493-502
• /
• 2020

Apigenin, a naturally occurring flavonoid, is known to exhibit significant anticancer activity. This study was designed to determine the effects of apigenin on two malignant mesothelioma cell lines, MSTO-211H and H2452, and to explore the underlying mechanism(s). Apigenin significantly inhibited cell viability with a concomitant increase in intracellular reactive oxygen species (ROS) and caused the loss of mitochondrial membrane potential (ΔΨm), and ATP depletion, resulting in apoptosis and necroptosis in monolayer cell culture. Apigenin upregulated DNA damage response proteins, including the DNA double strand break marker phospho (p)-histone H2A.X. and caused a transition delay at the G₂/M phase of cell cycle. Western blot analysis showed that apigenin treatment upregulated protein levels of cleaved caspase-3, cleaved PARP, p-MLKL, and p-RIP3 along with an increased Bax/Bcl-2 ratio. ATP supplementation restored cell viability and levels of DNA damage-, apoptosisand necroptosis-related proteins that apigenin caused. In addition, N-acetylcysteine reduced ROS production and improved ΔΨm loss and cell death that were caused by apigenin. In a 3D spheroid culture model, ROS-dependent necroptosis was found to be a mechanism involved in the anti-cancer activity of apigenin against malignant mesothelioma cells. Taken together, our findings suggest that apigenin can induce ROS-dependent necroptotic cell death due to ATP depletion through mitochondrial dysfunction. This study provides us a possible mechanism underlying why apigenin could be used as a therapeutic candidate for treating malignant mesothelioma.

• 한국발생생물학회지:발생과생식
• /
• 제14권3호
• /
• pp.185-197
• /
• 2010

The reproduction of animals is a means to maintain their species. The golden hamsters are seasonal breeders, and their reproduction is regulated by the photoperiod. The shifts of the sexual functions imply the changes of the protein expression, and those are reflected by the proteome. Thus the present study was to examine via two dimensional polyacrylamide gel electrophoresis (2d PAGE), the physiological changes and the alterations of protein expressions in hypothalamus upon the reproductive situation related to the pineal gland. Among the pineal intact animals, the reproductive functions were sustained in animals housed in long photoperiod (LP) but regressed in animals housed in short photoperiod. And those pinealectomized animals showed high sexual activities regardless of photoperiod. Ultimately they were branched into dichotomy, sexually active and inactive animals. Apart from the changes of physiological parameters upon the reproductive conditions, there were obvious differences in proteins extracted from the hypothalamus. The hypothalamus of LP animals presented high levels of enzymes which are involved in the production of energy, glycolysis and Krebs cycle. The increased energy might be related to the GnRH synthesis in hypothalamus and indirectly to the constant cell divisions in spermatogenesis. Taken together, the impacts of the photoperiodic information controlling reproduction could be observed through 2d PAGE. Therefore, the present results suggest the potential of biomarkers collectively to diagnose the fertility and the infertility by way of proteomics in organs with regard to the reproductive system, further could be applied to diagnose various diseases.

• 한국수산과학회지
• /
• 제44권3호
• /
• pp.216-224
• /
• 2011

Capsosiphon fulvescens is well-known green sea algae that, in recent years, has been proposed as a potential anticancer drug. In this study, we found that C. fulvescens glycoprotein (Cf-GP) had pro-apoptotic effects on human gastric carcinoma cells. By SDS-PAGE, we confirmed that C. fulvescens extract contained a glycoprotein. Using H33342 staining, we found that the Cf-GP caused cell death in a does-dependent manner, while an MTS assay showed decreased cellular viability due to induction of apoptosis. To determine the effect of Cf-GP on apoptosis-related cellular events, cells were treated with Cf-GP and the expression of several apoptosis-related protein was determined by Western blotting. Our results indicate that Cf-GP activated both a caspase cascade and PARP, which is a substrate of caspase-3, caspase-8 and the Bcl-2 family proteins. In addition, we assessed caspase-3, and -8 activation and annexin V staining. Our results revealed a cell cycle arrest, itself leading to an increased percentage of sub-G1 cells. Our findings indicate that Cf-GP may be a source of bio-functional material with therapeutic effects on human gastrointestinal cancer.

• Journal of Yeungnam Medical Science
• /
• 제24권1호
• /
• pp.24-40
• /
• 2007

배경 : 골관절염은 단순 노화로 인한 질병이 아니라 연골대사의 이상으로 인한 기계적 그리고 생화학적 불안정성이 나타나는 질환이다. Transforming growth factor-${\beta}$와 같이 연골세포의 기능을 향상시키는 촉진인자가 있는 반면 Interleukin(IL)-1이나 Tumor necrosis factor-${\alpha}$는 염증반응을 증가시킨다. 이중 IL-1은 골관절염의 병인에서 가장 중요한 염증 유발 인자로 알려져 있으며 이에 대한 자료도 축적되고 있다. 이 논문의 목적은 IL-$1{\beta}$에 대한 human chondrosarcoma cell (SW1353)의 유전자 발현 양상을 파악하여 골관절염 병인의 이해를 증대시키는데 있다. 재료 및 방법 : Chondrosarcoma cell line (SW 1353)은 연골세포의 IL-$1{\beta}$를 통한 세포노화에 대한 유전자 조절을 실험실에서 연구하는데 유용한 것으로 알려져 있다. 골관절염에서 IL-1에 의한 전체적인 유전자 발현의 변화를 연구하고 분석하기 위해 SW1353을 각각 1시간, 6시간, 24시간동안 IL-1에 노출시킨후 각각 총 RNA를 정제하였다. 유전자 발현의 변화는 17k human cDNA microarray로 분석하였고 semiquantitative RT-PCR로 확인하였다. 결과 : Metallothioneins, matrix metalloproteinases, extracellular proteins, antioxidant protein, cytoskeletal proteins, cell cycle regulatory proteins, 세포성장과 세포 자멸사에 대한 단백질, signal protein, transcriptional factor를 포함한 1,200개 유전자에서 2배 이상의 변화가 관찰되었다. 이러한 변화는 초기 관절염에서 보이는 병리생리학적 변화와 상관관계가 있는 것으로 생각된다. 결론 : cDNA microarray 분석으로 유전자 발현의 의미있는 변화를 관찰하였으며 골관절염 발병기전에서 분자생물학적 변화에 대한 인식과 치료 목표를 정립하는데 대한 새로운 자료로서 가치가 있을 것으로 생각된다.

• 생명과학회지
• /
• 제28권11호
• /
• pp.1316-1320
• /
• 2018

녹차(GT) 추출물의 효과를 인간 유방암 유래 세포인 MCF-7 세포를 사용하여 조사 하였다. GT추출물의 세포 독성 효과를 MTT 방법을 사용하여 관찰한 결과, MCF-7 세포는 현저한 세포 독성 효과를 보였고, 이 독성 효과는 GT추출물 농도 의존적으로 증가하였다. p53과 관련 단백질인 p21/cip1과 CDK2의 연관성을 조사하기 위해 GT추출물 처리 후 웨스턴 분석법을 통해 이들 단백질의 발현을 조사하였다. GT추출물 처리 후, MCF-7 세포에서 p53 단백질의 양은 농도에 따라 현저하게 증가 하였다. p21/cip1 단백질의 발현은 낮은 농도의 GT추출물에서 증가되며, 고농도에서도 감소하지 않았다. 그러나 CDK2의 단백질의 양은 높은 농도의 GT추출물에서 CDK2 발현의 급격한 감소가 관찰되었다. 이 결과는 GT추출물의 처리는 MCF-7 세포에서 p53와 p21/cip1를 증가시켜, 그 결과로 활성화 된 p21/cip1는 CDK2의 발현을 억제 함을 나타내고 있다. GT추출물이 MCF-7 세포의 세포주기에 어떤 영향을 미치는지 확인하기 위하여 FACS 분석으로 관찰한 결과, MCF-7 세포에서 세포주기의 G1 단계가 점차 증가하는 결과를 보였다. 이 결과는 GT추출물의 유방암 세포에서의 항암 효과는 세포주기의 G1 단계에서 MCF-7 세포를 정지시키는 p53에 의해 조절된다는 사실을 명확하게 보여 주고 있다.

• Biomolecules & Therapeutics
• /
• 제31권2호
• /
• pp.200-209
• /
• 2023

Patients with non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) amplification or sensitive mutations initially respond to the tyrosine kinase inhibitor gefitinib, however, the treatment becomes less effective over time by resistance mechanism including mesenchymal-epithelial transition (MET) overexpression. A therapeutic strategy targeting MET and EGFR may be a means to overcoming resistance to gefitinib. In the present study, we found that picropodophyllotoxin (PPT), derived from the roots of Podophyllum hexandrum, inhibited both EGFR and MET in NSCLC cells. The antitumor efficacy of PPT in gefitinib-resistant NSCLC cells (HCC827GR), was confirmed by suppression of cell proliferation and anchorage-independent colony growth. In the targeting of EGFR and MET, PPT bound with EGFR and MET, ex vivo, and blocked both kinases activity. The binding sites between PPT and EGFR or MET in the computational docking model were predicted at Gly772/Met769 and Arg1086/Tyr1230 of each ATP-binding pocket, respectively. PPT treatment of HCC827GR cells increased the number of annexin V-positive and subG1 cells. PPT also caused G2/M cell-cycle arrest together with related protein regulation. The inhibition of EGFR and MET by PPT treatment led to decreases in the phosphorylation of the downstream-proteins, AKT and ERK. In addition, PPT induced reactive oxygen species (ROS) production and GRP78, CHOP, DR5, and DR4 expression, mitochondrial dysfunction, and regulated involving signal-proteins. Taken together, PPT alleviated gefitinib-resistant NSCLC cell growth and induced apoptosis by reducing EGFR and MET activity. Therefore, our results suggest that PPT can be a promising therapeutic agent for gefitinib-resistant NSCLC.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권20호
• /
• pp.8623-8629
• /
• 2014

Background: As an important component of the NDC80 kinetochore complex, NUF2 is essential for kinetochore-microtubule attachment and chromosome segregation. Previous studies also suggested its involvement in development of various kinds of human cancers, however, its expression and functions in human hepatocellular carcinoma (HCC) are still unclear. Materials and Methods: In the present study, we aimed to test the hypothesis that NUF2 is aberrant in human HCCs and associated with cell growth. Results: Our results showed significantly elevated expression of NUF2 in human HCC tissues compared to adjacent normal tissues, and high expression of NUF2 in HCC cell lines. Using lentivirus-mediated silencing of NUF2 in HepG2 human HCC cells, we found that NUF2 depletion markedly suppressed proliferation and colony formation capacity in vitro, and dramatically hampered tumor growth of xenografts in vivo. Moreover, NUF2 silencing could induce cell cycle arrest and trigger cell apoptosis. Additionally, altered levels of cell cycle and apoptosis related proteins including cyclin B1, Cdc25A, Cdc2, Bad and Bax were also observed. Conclusions: In conclusion, these results demonstrate that NUF2 plays a critical role in the regulation of HCC cell proliferation and apoptosis, indicating that NUF2 may serve as a potential molecular target for therapeutic approaches.

• BMB Reports
• /
• 제47권12호
• /
• pp.697-702
• /
• 2014

Recently, the interest in natural products for the treatment of cancer is increasing because they are the pre-screened candidates. In the present study, we demonstrate the therapeutic effect of celastrol, a triterpene extracted from the root bark of Chinese medicine on gastric cancer. The proliferation of AGS and YCC-2 cells were most sensitively decreased in six kinds of gastric cancer cell lines after the treatment with celastrol. Celastrol inhibited the cell migration and increased G1 arrest in cell-cycle populations in both cell lines. The treatment with celastrol significantly induced autophagy and apoptosis and increased the expression of autophagy and apoptosis-related proteins. We also found an increase in phosphorylated AMPK following a decrease in all phosphorylated forms of AKT, mTOR and S6K after the treatment with celastrol. Moreover, gastric tumor burdens were reduced in a dose-dependent manner by celastrol administration in a xenografted mice model. Taken together, celastrol distinctly inhibits the gastric cancer cell proliferation and induces autophagy and apoptosis.

• BMB Reports
• /
• 제32권4호
• /
• pp.379-384
• /
• 1999

Taxol, a natural product with significant anti-tumor activity, stabilizes microtubules and arrests cells in the G2/M phase of the cell cycle. It has been reported that taxol has additional effects on the cell such as an increase in tyrosine phosphorylation of proteins and activation of mitogen-activated protein (MAP) kinase. This phosphorylated kinase translocates into the nucleus and phosphorylates its substrate c-jun, c-fos, ATF2, and ATF3. The MAP kinase family is comprised of key regulatory proteins that control the cellular response to both proliferation and stress signals. First examination was cytotoxicity and apoptosis-induced concentration with paclitaxel in HeLa cell. A half-maximal inhibition of cell proliferation ($IC_{50}$) occurred at 13 nM paclitaxel. When DNA fragmentation was analyzed by agarose gel electrophoresis, a nucleosomal ladder became evident 24 h after a taxol (50 nM) addition to the cells. In addition, an apoptotic body was detected by electron microscopy. Taxol-treated cells were arrested at the S phase at 10 nM. Treatment of 50 nM taxol activated the extracellular signal-regulated protein kinase (ERK1), and a fraction of the activated MAP kinases entered the nucleus. It was also discovered that nucleus substrates c-jun was phosphorylated and activated in the cell. The activated ERK1 could subsequently translocate into the nucleus and phosphorylate its substrate c-jun as well. This study suggests that taxol-induced apoptosis might be related with signal transduction via MAP kinases.

• Asian Pacific Journal of Cancer Prevention
• /
• 제15권5호
• /
• pp.2345-2351
• /
• 2014

Lysyl oxidase-like 2 (LOXL2), a member of the lysyl oxidase (LOX) family, is a copper-dependent enzyme that catalyzes oxidative deamination of lysine residues on protein substrates. LOXL2 was found to be overexpressed in esophageal squamous cell carcinoma (ESCC) in our previous research. We later identified a LOXL2 splicing variant LOXL2-delta72 and we overexpressed LOXL2-delta72 and its wild type counterpart in ESCC cells following microarray analyses. First, the differentially expressed genes (DEGs) of LOXL2 and LOXL2-delta72 compared to empty plasmid were applied to generate protein-protein interaction (PPI) sub-networks. Comparison of these two sub-networks showed hundreds of different proteins. To reveal the potential specific roles of LOXL2- delta72 compared to its wild type, the DEGs of LOXL2-delta72 vs LOXL2 were also applied to construct a PPI sub-network which was annotated by Gene Ontology. The functional annotation map indicated the third PPI sub-network involved hundreds of GO terms, such as "cell cycle arrest", "G1/S transition of mitotic cell cycle", "interphase", "cell-matrix adhesion" and "cell-substrate adhesion", as well as significant "immunity" related terms, such as "innate immune response", "regulation of defense response" and "Toll signaling pathway". These results provide important clues for experimental identification of the specific biological roles and molecular mechanisms of LOXL2-delta72. This study also provided a work flow to test the different roles of a splicing variant with high-throughput data.