• 제목/요약/키워드: P19 cells

검색결과 826건 처리시간 0.027초

Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer

  • Xu, Rong;Guo, Long-Jiang;Xin, Jun;Li, Wen-Mao;Gao, Yan;Zheng, You-Xian;Guo, You-Hong;Lin, Yang-Jun;Xie, Yong-Hua;Wu, Ya-Qing;Xu, Rui-An
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6557-6562
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    • 2013
  • Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

Shikonin Modulates Cell Proliferation by Inducing Apoptosis in LLC Cells via MAPK Regulation and Caspase Activation

  • 이수진;김성훈
    • 동의생리병리학회지
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    • 제19권2호
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    • pp.501-507
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    • 2005
  • Shikonin is a chemically characterized component of traditional herbal medicine, the root of Lithospermum erythrorhizon and has been shown to possess antitumor activities. Here we investigated anticancer potential of shikonin and its possible mechanism of action in LLC cells. Shikonin inhibited the proliferation of LLC cells in a concentration-dependent manner. It was also demonstrated that shikonin induced apoptosis in LLC cells by Annexin V staining and TUNEL staining analysis. Shikonin treatment was caused that decrease of Bcl-2, activation of caspases and cleavage of PARP. And shikonin also induced that the activation of mitogen-activated protein kinases (MAPKs), such as extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38. Interestingly, the cell proliferation inhibition induced by shikonin was recovered by specific inhibitors of JNK and p38 but the inhibitor of MEK, the upstream kinase of ERK, did not recover. Additionally, shikonin administration at doses of 5 mg/kg in C57BL/6 mice strongly inhibited the primary tumor growth of LLC. Taken together, these results suggest that shikonin may suppress LLC cell proliferation by inducing an apoptotic process via activation of caspases and MAPKs

Effects of Gastric Cancer Cells on the Differentiation of Treg Cells

  • Hu, Jing-Lan;Yang, Zhen;Tang, Jian-Rong;Fu, Xue-Qin;Yao, Lan-Jie
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권8호
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    • pp.4607-4610
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    • 2013
  • The aim of this study was evaluated the prevalence of Treg cells in peripheral blood in patients with gastric cancer, and investigate the effect of gastric cancer cells on their differentiation. ELISA was employed to assess the concentrations of TGF-${\beta}$ and IL-10 in gastric cancer patients' serum. Then, mouse gastric cancer cells were co-cultured with T lymphocytes or T lymphocytes + anti-TGF-${\beta}$. Flow cytometric analysis and RT-PCR were then performed to detect Treg cells and TGF-${\beta}$ and IL-10 expression in gastric cancer cells. Our data showed that the expression of TGF-${\beta}$ and IL-10 in the patients with gastric cancer was increased compared to the case with healthy donors. The population of Treg cells and the expression levels of TGF-${\beta}$ and IL-10 in the co-culture group were much higher than in the control group (18.6% vs 9.5%) (P<0.05). Moreover, the population of Treg cells and the expression levels of TGF-${\beta}$ and IL-10 in the co-culture systerm were clearly decreased after addition of anti-TGF-${\beta}$ (7.7% vs 19.6%) (P<0.01). In conclusion, gastric cancer cells may induce Treg cell differentiation through TGF-${\beta}$, and further promote immunosuppression.

COVID-19 Vaccination Alters NK Cell Dynamics and Transiently Reduces HBsAg Titers Among Patients With Chronic Hepatitis B

  • Hyunjae Shin;Ha Seok Lee;Ji Yun Noh;June-Young Koh;So-Young Kim;Jeayeon Park;Sung Won Chung;Moon Haeng Hur;Min Kyung Park;Yun Bin Lee;Yoon Jun Kim;Jung-Hwan Yoon;Jae-Hoon Ko;Kyong Ran Peck;Joon Young Song;Eui-Cheol Shin;Jeong-Hoon Lee
    • IMMUNE NETWORK
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    • 제23권5호
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    • pp.39.1-39.15
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    • 2023
  • Coronavirus disease 2019 (COVID-19) vaccination may non-specifically alter the host immune system. This study aimed to evaluate the effect of COVID-19 vaccination on hepatitis B surface Ag (HBsAg) titer and host immunity in chronic hepatitis B (CHB) patients. Consecutive 2,797 CHB patients who had serial HBsAg measurements during antiviral treatment were included in this study. Changes in the HBsAg levels after COVID-19 vaccination were analyzed. The dynamics of NK cells following COVID-19 vaccination were also examined using serial blood samples collected prospectively from 25 healthy volunteers. Vaccinated CHB patients (n=2,329) had significantly lower HBsAg levels 1-30 days post-vaccination compared to baseline (median, -21.4 IU/ml from baseline), but the levels reverted to baseline by 91-180 days (median, -3.8 IU/ml). The velocity of the HBsAg decline was transiently accelerated within 30 days after vaccination (median velocity: -0.06, -0.39, and -0.04 log10 IU/ml/year in pre-vaccination period, days 1-30, and days 31-90, respectively). In contrast, unvaccinated patients (n=468) had no change in HBsAg levels. Flow cytometric analysis showed that the frequency of NK cells expressing NKG2A, an NK inhibitory receptor, significantly decreased within 7 days after the first dose of COVID-19 vaccine (median, -13.1% from baseline; p<0.001). The decrease in the frequency of NKG2A+ NK cells was observed in the CD56dimCD16+ NK cell population regardless of type of COVID-19 vaccine. COVID-19 vaccination leads to a rapid, transient decline in HBsAg titer and a decrease in the frequency of NKG2A+ NK cells.

Analysis of Somatostatin-Secreting Gastric Delta Cells according to Upper Abdominal Symptoms and Helicobacter pylori Infection in Children

  • Kim, Dong-Uk;Moon, Jin-Hwa;Lee, Young-Ho;Paik, Seung Sam;Kim, Yeseul;Kim, Yong Joo
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • 제23권3호
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    • pp.243-250
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    • 2020
  • Purpose: Gastric delta cells (D-cells), which are somatostatin-secreting cells, are the main paracrine inhibitor of acid secretion. The number of D-cells was studied in children presenting with upper gastrointestinal (UGI) disease. Methods: We retrospectively investigated the number of D-cells in the gastric body and antrum through immunofluorescence examinations according to symptoms, endoscopic findings, and Helicobacter pylori infection in 75 children who visited Hanyang University Hospital Pediatrics. Results: The mean patient age was 12.2±3.3 years. The male-to-female ratio was 1:1.4. The mean D-cell number per high-power field in the antrum and body was 20.5 and 12 in children with substernal pain, 18.3 and 10.3 in vomiting, 22.3 and 6 in diarrhea, and 9.3 and 6 in abdominal pain, respectively (p>0.05). According to endoscopic findings, the mean D-cell number in the antrum and body was 14.3 and 6 with gastritis, 14 and 9.3 with reflux esophagitis, 16.7 and 8.7 with duodeno-gastric reflux, 19.3 and 12.7 with gastric ulcer, 16 and 13.7 with duodenitis, and 12.3 and 4 with duodenal ulcer, respectively (p>0.05). The D-cell number in the gastric body was 2.7 and 8.7 in children with current H. pylori infection and non-infected children, respectively (p=0.01), while those in the antrum were 15.5 and 14, respectively, with no statistical significance. Conclusion: The D-cell number was lower in the gastric body of children with current H. pylori infection. Further studies concerning peptide-secreting cells with a control group would provide information about the pathogenic pathways of UGI disorder.

Paricalcitol attenuates indoxyl sulfate-induced apoptosis through the inhibition of MAPK, Akt, and NF-κB activation in HK-2 cells

  • Park, Jung Sun;Choi, Hoon In;Bae, Eun Hui;Ma, Seong Kwon;Kim, Soo Wan
    • The Korean journal of internal medicine
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    • 제34권1호
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    • pp.146-155
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    • 2019
  • Background/Aims: Indoxyl sulfate (IS) is a uremic toxin and an important causative factor in the progression of chronic kidney disease. Recently, paricalcitol (19-nor-1,25-dihydroxyvitamin D2) was shown to exhibit protective effects in kidney injury. Here, we investigated the effects of paricalcitol treatment on IS-induced renal tubular injury. Methods: The fluorescent dye 2',7'-dichlorofluorescein diacetate was used to measure intracellular reactive oxygen species (ROS) following IS administration in human renal proximal tubular epithelial (HK-2) cells. The effects of IS on cell viability were determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays and levels of apoptosis-related proteins (Bcl-2-associated protein X [Bax] and B-cell lymphoma 2 [Bcl-2]), nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) p65, and phosphorylation of mitogen-activated protein kinase (MAPK) and protein kinase B (Akt) were determined by semiquantitative immunoblotting. The promoter activity of $NF-{\kappa}B$ was measured by luciferase assays and apoptosis was determined by f low cytometry of cells stained with f luorescein isothiocyanate-conjugated Annexin V protein. Results: IS treatment increased ROS production, decreased cell viability and induced apoptosis in HK-2 cells. IS treatment increased the expression of apoptosis-related protein Bax, decreased Bcl-2 expression, and activated phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt. In contrast, paricalcitol treatment decreased Bax expression, increased Bcl-2 expression, and inhibited phosphorylation of MAPK, $NF-{\kappa}B$ p65, and Akt in HK-2 cells. $NF-{\kappa}B$ promoter activity was increased following IS, administration and was counteracted by pretreatment with paricalcitol. Additionally, flow cytometry analysis revealed that IS-induced apoptosis was attenuated by paricalcitol treatment, which resulted in decreased numbers of fluorescein isothiocyanate-conjugated Annexin V positive cells. Conclusions: Treatment with paricalcitol inhibited IS-induced apoptosis by regulating MAPK, $NF-{\kappa}B$, and Akt signaling pathway in HK-2 cells.

Cloning and Characterization of the Zeaxanthin Glucosyltransferase Gene (crtX) from the Astaxanthin-Producing Marine Bacterium, Paracoccus haeundaensis

  • Seo, Yong-Bae;Choi, Seong-Seok;Nam, Soo-Wan;Lee, Jae-Hyung;Kim, Young-Tae
    • Journal of Microbiology and Biotechnology
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    • 제19권12호
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    • pp.1542-1546
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    • 2009
  • Zeaxanthin glucosyltransferase (CrtX) mediates the formation of zeaxanthin to zeaxanthin diglucoside. Here, we report cloning of the crtX gene responsible for zeaxanthin diglucoside biosynthesis from Paracoccus haeundaensis and the production of the corresponding carotenoids in transformed cells carrying this gene. An expression plasmid containing the crtX gene (pSTCRT-X) was constructed, and Escherichia coli cells containing this plasmid produced the recombinant protein of approximately 46 kDa. Biosynthesis of zeaxanthin diglucoside was obtained when the plasmid pSTCRT-X was co-transformed into E. coli containing the pET-44a(+)-CrtEBIYZ carrying crtE, crtB, crtI, crtY, and crtZ genes required for zeaxanthin $\beta$-D-diglucoside biosynthesis.

원발성 폐선암환자의 말초혈액에서 분리한 CD1c+ 골수성 수지상 세포에서의 CD40, CD86 및 HLA-DR의 발현 (Expression of CD40, CD86, and HLA-DR in CD1c+ Myeloid Dendritic Cells Isolated from Peripheral Blood in Primary Adenocarcinoma of Lung)

  • 강문철;강창현;김영태;김주현
    • Journal of Chest Surgery
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    • 제43권5호
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    • pp.499-505
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    • 2010
  • 배경: CD1-restricted T 세포는 동물모델실험에서 항암면역에 있어서 매우 중요한 역할을 한다는 보고가 있다. 이를 임상에서 확인하기 위해 원발성 폐선암환자의 말초혈액에 존재하는 CD1c+ 골수성 수지상 세포의 수 및 수지상 세포에서의 동시자극 표지자 중 주요 표지자인 CD40, CD86 및 HLA-DR의 발현 양상을 확인하고자 하였다. 대상 및 방법: 영상의학적으로 원발성 폐암이 의심되거나 조직학적으로 폐암으로 확진되어 외과적 폐절제 또는 생검을 시행한 총 24명의 환자를 원발성 폐선암 환자군(Primary adenocarinoma of lung, PACL)과 원발성 편병상피세포폐암군(Primary squamous cell carcinoma of lung, PSqCL) 및 양성폐질환군(Benign lung disease, BLD)군으로 나누고, 말초혈액 20 mL을 채취하여 수지상 세포를 분리하고 단일클론 항체를 이용하여 CD1c+ 골수성 세포의 수 및 수지상 세포에서의 동시자극 표지자 중 주요 표지자들의 발현정도를 유체세포측정기를 이용하여 확인하고 두 군 간의 통계적 차이 유무를 확인하였다. 결과: CD1c+ 골수성 수지상 세포(CD1c+CD19-)의 수는 PACL, PSqCL 및 BLD군 모두 그 차이가 모두 통계적으로 유의한 차이는 없었다. 공동자극표지자의 발현은 PACL군과 BLD군에서는 CD40의 발현이 p-value=0.171, CD86의 발현이 p-value=0.037과 HLA-DR의 발현이 p-value=0.036로 PACL군에서 감소한 것을 통계학적으로 확인할 수 있었다. PACL군과 PSqCL군 간에는 CD40의 발현이 p-value=0.319, CD86의 발현이 p-value= 0.036과 HLA-DR의 발현이 p-value=0.085로 PACL군에서 공동표지자의 발현이 감소한 양상이나 통계적 유의성은 CD86의 발현 외에는 없었다. PSqCL군과 BLD군간에는 공동표지자의 발현에 통계적 차이는 없었다. PACL군과 비선암군간에는 CD40의 발현이 p-value=0.005, CD86의 발현이 p-value=0.013과 HLA-DR의 발현이 p-value=0.004로 PACL군에서 공동표지자의 발현이 감소한 것을 통계학적으로 확인할 수 있었다. 결론: CD1c+ 골수성 수지상 세포에서 원발성 폐선암 환자에서 주요 표지자의 발현이 감소한 것을 확인할 수 있었으며, 이는 원발성 폐선암 환자에서 주요 표지자의 발현의 감소가 면역회피기전의 하나일 수 있는 가능성 및 수지상 세포에 의하여 유발되는 면역반응은 수지상 세포의 표지자의 발현 정도에 의해 결정된다는 가능성을 제시한다.

산화물구리 기반 이종접합형 태양전지의 후열처리효과 (Effect of Post-annealing Treatment on Copper Oxide based Heterojunction Solar Cells)

  • 김상모;정유섭;김경환
    • 반도체디스플레이기술학회지
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    • 제19권2호
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    • pp.55-59
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    • 2020
  • Copper Oxide (CuO) films were deposited on the n-type silicon wafer by rf magnetron sputtering for heterojunction solar cells. And then the samples were treated as a function of the annealing temperature (300-600℃) in a vacuum. Their electrical, optical and structural properties of the fabricated heterojunction solar cells were then investigated and the power conversion efficiencies (PCE) of the fabricated p-type copper oxide/n-type Si heterojunction cells were measured using solar simulator. After being treated at temperature of 500℃, the solar cells with CuO film have PCE of 0.43%, Current density of 5.37mA/㎠, Fill Factor of 39.82%.

Anti-stress Effects of Ethanol Extract of Ziziphus jujuba Against Corticosterone-Induced Apoptosis in PC12 Cells

  • Da Hye Song;Yu Jin Choi
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2022년도 추계학술대회
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    • pp.119-119
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    • 2022
  • The coronavirus disease 2019 (COVID-19) pandemic may be stressful for people. Public health actions, such as social distancing, can make people feel isolated and lonely and can increase stress and anxiety. As a result, there is a growing interest towards various materials to relieve stress. Thus, the present study aimed to investigate the anti-stress effects of ethanol extract of Ziziphus jujuba in PC12 cells treated with corticosterone and its underling mechanisms. Furthermore, the viability of the cells, the apoptosis of the cells, the level of phosphorylation of extracellular signal-regulated kinases (p-ERKs) expression were measured by MTT assay, LDH assay, Hoechst staining assay and western blotting. Our results showed that the extract of Ziziphus jujuba reversed corticosterone-induced damage in PC12 cells, which increased cell viability, decreased LDH release, and attenuated corticosterone-induced apoptosis as compared with the corticosterone-treated group. Therefore, these data suggest that the extract of Ziziphus jujuba could be a good candidate for development as a functional food supplement in the improve the anti-stress effect.

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