• Journal of Microbiology and Biotechnology
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• 제26권2호
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• pp.287-294
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• 2016

The effect of kaempferol (3,5,7,4-tetrahydroxyflavone), a flavonoid compound that was identified in barnyard millet (Echinochloa crus-galli var. frumentacea) grains, on G₂-checkpoint and apoptotic pathways was investigated in human acute leukemia Jurkat T cell clones stably transfected with an empty vector (J/Neo) or a Bcl-xL expression vector (J/Bcl-xL). Exposure of J/Neo cells to kaempeferol caused cytotoxicity and activation of the ATM/ATR-Chk1/Chk2 pathway, activating the phosphorylation of p53 (Ser-15), inhibitory phosphorylation of Cdc25C (Ser-216), and inactivation of cyclin-dependent kinase 1 (Cdk1), with resultant G₂-arrest of the cell cycle. Under these conditions, apoptotic events, including upregulation of Bak and PUMA levels, Bak activation, mitochondrial membrane potential (Δψm) loss, activation of caspase-9, -8, and -3, anti-poly (ADP-ribose) polymerase (PARP) cleavage, and accumulation of apoptotic sub-G₁ cells, were induced without accompanying necrosis. However, these apoptotic events, except for upregulation of Bak and PUMA levels, were completely abrogated in J/Bcl-xL cells overexpressing Bcl-xL, suggesting that the G₂-arrest and the Bcl-xL-sensitive mitochondrial apoptotic events were induced, in parallel, as downstream events of the DNA-damage-mediated G₂-checkpoint activation. Together these results demonstrate that kaempferol-mediated antitumor activity toward Jurkat T cells was attributable to G₂-checkpoint activation, which caused not only G₂-arrest of the cell cycle but also activating phosphorylation of p53 (Ser-15) and subsequent induction of mitochondria-dependent apoptotic events, including Bak and PUMA upregulation, Bak activation, Δψm loss, and caspase cascade activation.

• Journal of Oral Medicine and Pain
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• 제30권2호
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• pp.231-238
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• 2005

항암제의 연구는 화학물질에 민감한 암세포를 죽음에 이르게 하는 세포자멸사와 같은 다양한 세포기능에 초점을 맞추어 왔다. 그러나 약물이 유도한 세포의 죽음에 있어서 핵심적인 분자적 기작은 아직 잘 이해되지 않고 있다. Etoposide는 폐암과 고환암에 사용되는 항암제로서, 본 연구는 etoposide가 사람구강편평상피암종세포(OSC9)에도 세포독성효과와 세포자멸사를 일으키는지를 알아보기 위해 실행하였다. 이 실험에서 etoposide는 농도와 시간 의존적으로 OSC9 세포의 생존율를 현저하게 저해시켰다. TUNEL 염색과 Hoechst 염색을 이용한 핵의 형태학적 관찰에서는 etoposide에 의해 핵이 응축되고 분절되었다. p53의 발현은 48 시간에 증가했으며, etoposide 처리로 인해 caspase-3의 활성을 관찰할 수 있었으며, 그 기질에 해당되는 PARP 단백질은 116-kDa과 89-kDa으로 분절되었다. 위의 결과들은 OSC9 세포에서 etoposide가 유도한 세포자멸사는 caspase-3의 활성과 관련됨을 설명하고 있다.

• Journal of Biomedical and Translational Research
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• 제19권4호
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• pp.116-124
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• 2018

Korean Native Pig (KNP) has a uniform black coat color, excellent meat quality, white colored fat, solid fat structure and good marbling. However, its growth performance is low, while the western origin Yorkshire pig has high growth performance. To take advantage of the unique performance of the two pig breeds, we raised crossbreeds (KNP ${\times}$ Yorkshire to make use of the heterotic effect. We then analyzed the liver transcriptome as it plays an important role in fat metabolism. We sampled at two stages: 10 weeks and at 26 weeks. The stages were chosen to correspond to the change in feeding system. A total of 16 pigs (8 from each stage) were sampled and RNA sequencing was performed. The reads were mapped to the reference genome and differential expression analysis was performed with edgeR package. A total of 324 genes were found to be significantly differentially expressed (${\left|log2FC\right|}$ > 1 & q < 0.01), out of which 180 genes were up-regulated and 144 genes were down-regulated. Principal Component Analysis (PCA) showed that the samples clustered according to stages. Functional annotation of significant DEGs (differentially expressed genes) showed that GO terms such as DNA replication, cell division, protein phosphorylation, regulation of signal transduction by p53 class mediator, ribosome, focal adhesion, DNA helicase activity, protein kinase activity etc. were enriched. KEGG pathway analysis showed that the DEGs functioned in cell cycle, Ras signaling pathway, p53 signaling pathway, MAPK signaling pathway etc. Twenty-nine transcripts were also part of the DEGs, these were predominantly Cys2His2-like fold group (C2H2) family of zinc fingers. A protein-protein interaction (PPI) network analysis showed that there were three highly interconnected clusters, suggesting an enrichment of genes with similar biological function. This study presents the first report of liver tissue specific gene regulation in a cross-bred Korean pig.

• 한국방사선학회논문지
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• 제17권3호
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• pp.465-472
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• 2023

본 연구는 혈관질환을 예측하는 데 있어서 일차 선별검사로서 경동맥 초음파 검사와 혈액학적 검사를 실시한 환자를 대상으로 경동맥 내중막 두께의 변화를 확인하고 죽상경화반의 발현에 위험인자들이 미치는 영향을 알아보고자 하였다. 일개 병원에서 건강검진을 목적으로 내원하여 경동맥 초음파 검사를 실시한 건강한 성인 469명을 대상으로 후향적으로 분석하였다. 그 결과 경동맥 내중막 두께, 연령, 체질량지수, 허리둘레, 수축기 혈압, 총콜레스테롤, 고비중지단백 콜레스테롤, 저비중지단백 콜레스테롤, 공복혈당이 죽상경화반의 유의한 예측인자로 분석되었다(p<0.001). ROC curve 분석을 통해 결정된 죽상경화반 위험인자들의 Cut off value를 기준으로 위험비를 산출하였으며 경동맥 내중막 두께는 8.06배, 연령은 7.53배, 허리둘레는 3.97배, 공복혈당은 2.02배 순으로 높게 나타났다. 따라서 본 연구에서는 죽상경화반의 유무에 영향을 주는 임상학적 위험요인들의 한국인 기준치를 마련할 수 있었으며, 초음파를 이용한 경동맥의 관찰은 심뇌혈관 질환을 조기에 진단하거나 예측하는데 도움이 될 것으로 생각된다.

• 한국간담췌외과학회지
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• 제26권1호
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• pp.58-68
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• 2022

Backgrounds/Aims: Mechanisms for the development of hepatocellular carcinoma (HCC) in hepatitis B virus (HBV)-infected patients remain unclear. The aim of the present study was to identify genes and pathways involved in the development of HBV-associated HCC. Methods: The GSE121248 gene dataset, which included 70 HCCs and 37 adjacent liver tissues, was downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) in HCCs and adjacent liver tissues were identified. Gene ontology and Kyoto Encyclopedia of Genes and Genome pathway enrichment analyses were then performed. Results: Of 134 DEGs identified, 34 were up-regulated and 100 were down-regulated in HCCs. The 34 up-regulated DEGs were mainly involved in nuclear division, organelle fission, spindle and midbody formation, histone kinase activity, and p53 signaling pathway, whereas the 100 down-regulated DEGs were involved in steroid and hormone metabolism, collagen-coated extracellular matrix, oxidoreductase activity, and activity on paired donors, including incorporation or reduction of molecular oxygen, monooxygenase activity, and retinol metabolism. Analyses of protein-protein interaction networks with a high degree of connectivity identified significant modules containing 14 hub genes, including ANLN, ASPM, BUB1B, CCNB1, CDK1, CDKN3, ECT2, HMMR, NEK2, PBK, PRC1, RACGAP1, RRM2, and TOP2A, which were mainly associated with nuclear division, organelle fission, spindle formation, protein serine/threonine kinase activity, p53 signaling pathway, and cell cycle. Conclusions: This study identified key genes and carcinogenic pathways that play essential roles in the development of HBV-associated HCC. This may provide important information for the development of diagnostic and therapeutic targets for HCC.

• 한국식품과학회지
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• 제53권1호
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• pp.34-39
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• 2021

본 연구에서는 천연유래 물질로부터 apoptosis를 유도하여 항암 활성이 있는지에 관한 실험으로 사람 난소암 세포주인 OVCAR-3 세포에 함초의 유기용매별 추출물을 처리하여 결과를 확인하였다. MTS 측정으로 세포 생존율을 확인한 결과 DCM 분획물에서 농도별로 유의적인 세포수의 감소를 보였으며, annexin V/FITC-PI 염색에 의해 apoptosis 유도로 세포가 사멸함을 확인하였다. DCM 분획물 처리는 세포주기에서 Sub-G1기의 증가로 세포증식이 억제됨을 보여준다. 세포의 내인성 경로에 관여하는 Bcl-2 family에 속하는 Bax, Bak, Bcl-2, Bcl-xL의 상호작용을 mRNA 수준에서 확인한 결과, DCM 분획물처리에서 Bax, Bak가 증가하고, Bcl-2의 감소를 동반하여 세포사멸의 신호전달 경로가 진행됨을 확인할 수 있었다. 본 연구 결과를 바탕으로 함초를 이용하여 여성의 난소암에 예방적 기능성 식품을 개발할 수 있을 것으로 판단되며, 함초의 DCM 분획물에 대해 깊이 있는 심층적 연구가 요구된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권1호
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• pp.14-19
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• 2012

Introduction: This study was conducted to evaluate ssrA expression resulting from adaptation of Escherichia coli (E. coli) to oral pathogens through signal exchange. Materials and Methods: Human cell lines Hep2 and HT29, wild-type E. coli (WT K-12), ssrA knock-out E. coli (${\Delta}K$-12), and Scleropages aureus (S. aureus) were used. A single culture consisting of Hep2, HT29, WT K-12, and ${\Delta}K$-12, and mixed cultures consisting of Hep2 and WT K-12, Hep2 and ${\Delta}K$-12, WT K-12 and S. aureus, ${\Delta}K$-12 and S. aureus, and Hep2, WT K-12, and S. aureus were prepared. For HT29, a mixed culture was prepared with WT K-12 and with WT K-12 and S. aureus. Total RNA was extracted from each culture with the resulting expression of ssrA, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$), and p53 was evaluated by Reverse transcription polymerase chain reaction (RT-PCR). Results: The expression of ssrA in a single culture of WT K-12 was lower than that observed in the mixed culture of WT K-12 with S. aureus. Greater ssrA expression was observed in the mixed culture of WT K-12 with Hep2 than in the single culture of WT K-12. The expression of NF-${\kappa}B$ was higher in the mixed culture of Hep2 with ${\Delta}K$-12 than that in the mixed culture of Hep2 with WT K-12, and was lowest in the single culture of Hep2. The expression of ssrA was higher in the mixed culture of WT K-12 with Hep2 and S. aureus than in the mixed culture of WT K-12 with Hep2. Conclusion: These results suggest that ssrA plays an important role in the mechanism of E. coli adaptation to a new environment.

• 대한화장품학회지
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• 제30권3호
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• pp.393-397
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• 2004

L-Carnitine $({\beta}-hydroxy-\gamma-trimethyl-ammoniumbutyric{\;}acid)$은 분자량이 적은 수용성 분자로서 세포 내 지방 대사에서 중요한 역할을 수행한다. 지방산의 운반 분자인 아실-코에이(acyl-CoA)가 미토콘드리아의 세포막을 투과하지 못하기 때문에 지방산은 CoA로부터 카르니틴으로 운반되어 미토콘드리아에서 작용한다. 노화와 연관된 L-carnitine의 기능을 확인하기 위하여 MMP inhibition assay와 자외선 조사에 의해 유도된 MMP 발현에 대한 영향을 확인하였다. MMP inhibition assay는 콜라겐을 이용한 형광분석법을 실시하였고 자외선 조사에 의해 유도된 MMP 발현양은 ELISA로 정량하였으며 그 활성은 젤라틴 기질 zymography로 확인하였고 MMP mRNA 발현양은 RT-PCR ELISA로 확인하였다. 또한, 사람을 이용한 임상 실험을 통하여 주름 개선 효과를 평가하였다. L-carnitine은 농도 의존적으로 MMP 저해 활성을 나타났으며 $IC_{50}$값은 2.45 mM이었으며 자외선 조사에 의해 발현된 MMP 활성을 강하게 저해하였다. 자외선 조사에 의해 발현되는 MMP에 대해 단백질의 양적인 변화는 $40\%$ 정도 감소되었으며 L-carnitine 처리에 의해 농도 의존적으로 MMP mRNA의 발현양은 감소되었다. 이러한 실험결과를 통하여 L-carnitine은 MMP 효소의 저해능 뿐만 아니라 자외선 조사에 의해 유도되는 MMP 단백질 발현과 mRNA 유전자 수준에서의 조절이 가능함을 확인하였다. 사람을 이용한 임상 실험에서는 $1\%$ 카르니틴을 함유하는 화장품을 약 3개월간 사용 후에는 유의적으로 주름 개선 효과를 확인하였다. 결론적으로 L-Carnitine은 광노화에 관여하는 MMP 활성과 발현 조절 메커니즘을 통하여 광손상에 대응하는 항노화 소재로서의 화장품에 매우 효과적이었음을 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권17호
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• pp.7633-7640
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• 2015

Background: Human papillomavirus (HPV) is the most common sexually transmitted infection worldwide and it is responsible for most cases of cervical uterine cancer. Although HPV infections of the cervix do not always progress to cancer, 90% of cervical cancer cases have been found to be associated with high risk HPV (HR-HPV) infection. HPV DNA testing is widely used, along with Papanicolaou (Pap) testing, to screen for cervical abnormalities. However, there are no data on the prevalence of genotype-specific HPV infections assessed by measuring HPV E6/E7 mRNA in women representative of the Chinese population across a broad age range. Materials and Methods: In the present study, we compared the results with the CervicGen HPV RT-qDx assay, which detects 16 HR-HPV genotypes (Alpha-9: HPV 16, 31, 33, 35, 52, and 58; Alpha-7: HPV 18, 39, 45, 51, 59, and 68; and Alpha-5, 6: HPV 53, 56, 66, and 69), and the REBA HPV-ID assay, which detects 32 HPV genotypes based on the reverse blot hybridization assay (REBA) for the detection of oncogenic HPV infection according to cytological diagnosis. We also investigated the prevalence and genotype distribution of HPV infection with a total of 324 liquid-based cytology samples collected in western Shandong province, East China. Results: The overall HPV prevalences determined by HPV DNA and HPV E6/E7 mRNA assays in this study were 79.9% (259/324) and 55.6% (180/324), respectively. Although the positivity of HPV E6/E7 mRNA expression was significantly lower than HPV DNA positivity, the HPV E6/E7 mRNA assay showed greater specificity than the HPV DNA assay (88.6% vs. 48.1%) in normal cytology samples. The prevalence of Alpha-9 (HPV 16, 31, 33, 35, 52, and 58) HPV infection among these women accounted for up to 80.3% and 76.1% of the high-grade lesions detected in the HPV mRNA and DNA tests, respectively. The HR-HPV genotype distribution, based on HPV DNA and E6/E7 mRNA expression by age group in patients with cytologically confirmed lesions, was highest in women aged 40 to 49 years (35.9% for cytologically confirmed cases, Pearson correlation r value=0.993, p<0.001) for high-grade lesions. Among the oncogenic HR-HPV genotypes for all age groups, there was little difference in the distribution of HPV genotypes between the HPV DNA (HPV -16, 53, 18, 58, and 33) and HPV E6/E7 mRNA (HPV -16, 53, 33, 58, and 18) assays. HPV 16 was the most common HPV genotype among women with high-grade lesions. Conclusions: Our results suggest that the HPV E6/E7 mRNA assay can be a sensitive and specific tool for the screening and investigation of cervical cancer. Furthermore, it may provide useful information regarding the necessity for early cervical cancer screenings and the development of additional effective HPV vaccines, such as one for HPV 53 and 58. Additionally, gaining knowledge of HPV distribution may also inform us about ecological changes in HPV after the vaccination.

• Animal cells and systems
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• 제3권1호
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• pp.53-58
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• 1999

The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2＋}$-channels in mouse follicular oocytes. Three types of voltage-dependent $Ca^{2＋}$-channels were shown to exist in the follicular oocytes for the first time, the P/Q-type $Ca^{2＋}$-channel, the N-type $Ca^{2＋}$-channel, and the L-type $Ca^{2＋}$-channel. Among proven $Ca^{2＋}$-channels distributions of the P/Q-type $Ca^{2＋}$-channel and L-type $Ca^{2＋}$-channel showed localized staining (clustered pattern) on the oolemma. The distribution of the P/Q-type $Ca^{2＋}$-channel showed all localized staining, and the range of localized staining was from 1 to 8 in staining intensity. As the staining intensity increased from 1 to 8, the number of localized staining decreased. The L-type $Ca^{2＋}$-channel are homogeneously stained (29.4%-54.2%), while some of them (around 28.7%-44.1%) showed localized staining on the oolemma. However, the rest of them showed no staining at all (17.1%- 26.5%). On the contrary, the N-type $Ca^{2＋}$-channel showed mostly homogeneous staining, while nonstaining oocytes were around 33.8%. The rest showed localized staining (10%). However, staining intensity was much weaker than those of the P/Q-type and L-type $Ca^{2＋}$-channel. In fact, the N-type $Ca^{2＋}$-channel has been known to exist only in neurons (from ectoderm origin), but it is unknown how the N-type $Ca^{2＋}$-channel exists in the follicular oocytes (from mesoderm origin). Further studies are needed to examine the expression of $Ca^{2＋}$-channels during the developmental stages of the oocytes.