• 제목/요약/키워드: proliferative capacity

검색결과 48건 처리시간 0.026초

Establishment and Characterization of Immortalized Human Dermal Papilla Cells Expressing Human Papillomavirus 16 E6/E7

  • Seonhwa Kim;Kyeong-Bae Jeon;Hyo-Min Park;Jinju Kim;Chae-Min Lim;Do-Young Yoon
    • Journal of Microbiology and Biotechnology
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    • 제34권3호
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    • pp.506-515
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    • 2024
  • Primary human dermal papilla cells (HDPCs) are often preferred in studies on hair growth and regeneration. However, primary HDPCs are limited by their reduced proliferative capacity, decreased hair induction potential, and extended doubling times at higher passages. To overcome these limitations, pTARGET vectors containing human papillomavirus16 (HPV16) E6/E7 oncogenes were transfected into HDPCs and selected using G-148 to generate immortalized cells here. HPV16 E6/E7 oncogenes were efficiently transfected into primary HDPCs. Immortalized HDPC showed higher proliferative activity than primary HDPC, confirming an increased proliferation rate. Expression of p53 and pRb proteins was downregulated by E6 and E7, respectively. E6/E7 expressing HDPC cells revealed that cyclin-dependent kinase (CDK) inhibitor p21 expression was decreased, while cell cycle-related genes and proteins (CDK2 and cyclin E) and E2F family genes were upregulated. Immortalized HDPCs maintained their responsiveness to Wnt/β-catenin pathway and hair follicle formation capability, as indicated by their aggregative properties and stemness. E6/E7 immortalized HDPCs may facilitate in vitro hair growth and regeneration studies.

세포배양삽입체계(Cell Culture Insert System)에서 중간엽 줄기세포(Mesenchymal Stem Cell)가 수지상세포(Dendritic Cell)의 활성화에 미치는 영향 (The Effect of Mesenchymal Stem Cells on the Activation of Dendritic Cells in the Cell Culture Insert System)

  • 김기원;박석영;이경복;김현수
    • IMMUNE NETWORK
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    • 제4권2호
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    • pp.88-93
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    • 2004
  • Background: Bone marrow mesenchymal stem cells (MSC) inhibit the immune response of lymphocytes to specific antigens and dendritic cells (DC) are professional antigenpresenting cells whose function is to present antigen to naive T-lymphocytes with high efficiency and play a central role in the regulation of immune response. We studied the effects of MSC on DC to evaluate the relationship between MSC and DC in transplantation immunology. Methods: MSC were expanded from the bone marrow and DC were cultured from peripheral blood mononuclear cells (PBMNC) of 6 myelogenous leukemia after achieving complete response. Responder cells isolated from PBMNC and lysates of autologous leukemic cells are used as tumor antigen. The effect of MSC on the DC was analyzed by immunophenotype properties of DC and by proliferative capacity and the amount of cytokine production with activated PBMNC against the allogeneic lymphocytes. Also, cytotoxicity tests against leukemic cells studied to evaluate the immunologic effect of MSC on the DC. Results: MSC inhibit the CD83 and HLA-class II molecules of antigen-loaded DC. The proliferative capacity and the amount of INF-$\gamma$ production of lymphocytes to allogeneic lymphocytes were decreased in DC co-cultured with MSC. Also the cytotoxic activity of lymphocytes against leukemic cells was decreased in DC co-cultured with MSC. Conclusion: MSC inhibit the activation and immune response of DC induced by allogeneic or tumor antigen.

Links between accelerated replicative cellular senescence and down-regulation of SPHK1 transcription

  • Kim, Min Kyung;Lee, Wooseong;Yoon, Gang-Ho;Chang, Eun-Ju;Choi, Sun-Cheol;Kim, Seong Who
    • BMB Reports
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    • 제52권3호
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    • pp.220-225
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    • 2019
  • We have identified a mechanism to diminish the proliferative capacity of cells during cell expansion using human adipose-derived stromal cells (hAD-SCs) as a model of replicative senescence. hAD-SCs of high-passage numbers exhibited a reduced proliferative capacity with accelerated cellular senescence. Levels of key bioactive sphingolipids were significantly increased in these senescent hAD-SCs. Notably, the transcription of sphingosine kinase 1 (SPHK1) was down-regulated in hAD-SCs at high-passage numbers. SPHK1 knockdown as well as inhibition of its enzymatic activity impeded the proliferation of hAD-SCs, with concomitant induction of cellular senescence and accumulation of sphingolipids, as seen in high-passage cells. SPHK1 knockdown-accelerated cellular senescence was attenuated by co-treatment with sphingosine-1-phosphate and an inhibitor of ceramide synthesis, fumonisin $B_1$, but not by treatment with either one alone. Together, these results suggest that transcriptional down-regulation of SPHK1 is a critical inducer of altered sphingolipid profiles and enhances replicative senescence during multiple rounds of cell division.

면역결핍 모델에서 β-1,3/1,6-glucan과 유산균을 이용한 in vivo 면역 활성 조절 효과 (Immunomodulatory effects of β-1,3/1,6-glucan and lactic acid bacteria in LP-BM5 murine leukemia viruses-induced murine acquired immune deficiency syndrome)

  • 김민수;김중수;류민정;김기홍;황권택
    • 한국식품저장유통학회지
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    • 제24권8호
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    • pp.1158-1167
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    • 2017
  • 본 연구에서는 LP-BM5 murine AIDS virus에 감염된 면역 결핍동물 모델을 이용하여 실험기간 10주 동안 ${\beta}$-1,3/1,6-glucan, 유산균 및 ${\beta}$-1,3/1,6-glucan+유산균의 식이가 면역활성에 미치는 영향을 조사하였다. 그 결과 LP-BM5 murine AIDS virus에 감염으로 면역능이 떨어진 T세포 증식능은 홍삼대조군과 비교하여도 유의적으로 감소된 T 세포증식능을 증가시키는 것으로 나타났고, B 세포 증식능은 감염 대조군에 비하여 유의적으로 증가된 B 세포 증식능을 감소되었다. cytokine 생성능에서는 Th1 type cytokine중에서 IL-2, IL-12, IL-15는 감염대조군에 비하여 분비량을 증가시키는 것을 확인할 수 있었고, IFN-${\gamma}$는 유산균과 ${\beta}$-1,3/1,6-glucan을 각각 처리군이 혼합처리군보다 증식능이 증가하였다. TNF-${\alpha}$는 감염대조군에 비하여 유의적으로 감소하였다. Th2 cytokine 들의 분비량 측정에서 IL-4, IL-6, IL-10 측정 결과 감염대조군에서 유의적으로 억제되어 Th1/Th2 type cytokine 발현을 조절하여 면역항상성을 유지하는 것으로 보였다. 면역글로블린 분비량측정에서 IgE, IgA, IgG 모두 감염대조군에 비하여 유의적으로 떨어지는 것으로 나타났다. 이로서LP-BM5 murine AIDS virus에 감염된 면역 결핍동물 모델에 ${\beta}$-1,3/1,6-glucan+유산균군을 혼합처리로 면역조절의 효능이 있음을 확인할 수 있었다.

Inhibitory Effects of Opuntia humifusa on 7, 12-Dimethyl-benz[a]anthracene and 12-O-tetradecanoylphorbol-13-acetate Induced Two-stage Skin Carcinogenesis

  • Lee, Jin-A;Jung, Bock-Gie;Lee, Bong-Joo
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권9호
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    • pp.4655-4660
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    • 2012
  • Opuntia humifusa, member of the Cactaceae family, was previously demonstrated to have radical scavenging, anti-inflammatory and anti-proliferative effects in in vitro models. It was suggested that O. humifusa could function in the prevention of carcinogenesis. To investigate the in vivo chemopreventive effect of O. humifusa, mice were fed a diet containing either 1% or 3% following 7, 12-dimethylbenz[a] anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) induction of skin carcinogenesis. Significant decrease in the numbers of papilloma and epidermal hyperplasia were observed in mice fed with O. humifusa, compared to the control group. O. humifusa also upregulated high total antioxidant capacity and level of phase II detoxifying enzyme such as superoxide dismutase and glutathione S-transferase activity in the skin. Lipid peroxidation activity level was measured in skin cytosol and significantly inhibited in 3% OH fed group compared to the control group. These results suggest that O. humifusa exerts chemopreventive effects on chemical carcinogenesis in mouse skin and that prevention effects are associated with reduction of oxidative stress via the modulation of cutaneous lipid peroxidation, enhancing of total antioxidant capacity especially in phase II detoxifying enzyme system and partial apoptotic influence.

The Clinical and Radiographic Features of Patients with Temporomandibular Joint Osteoarthritis: Comparison of Adolescents and Middle-Old Aged Koreans

  • Kim, Jin-Hwa;Ok, Soo-Min;Heo, Jun-Young;Kim, Kyung-Hee;Jeong, Sung-Hee;Ahn, Yong-Woo;Ko, Myung-Yun
    • Journal of Oral Medicine and Pain
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    • 제39권1호
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    • pp.2-9
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    • 2014
  • Purpose: The purpose of this study was to compare the clinical and radiographic features of temporomandibular joint (TMJ) osteoarthritis (OA) between adolescents and middle-old aged patients. Methods: The subjects were chosen among the patients who presented to the Department of Oral Medicine of Pusan National University Hospital and were diagnosed with TMJ OA by clinical exam, X-ray and cone-beam computed tomography (CBCT) from 2010 to 2011. We investigated 93 adolescent patients (12-19 years) and 53 middle-old aged (>45 years) patients who observed the erosive bony changes in TMJ. CBCT scans were retaken at intervals at an average of 8 months. Results: The adolescent patients showed unilateral degenerative changes more often, and the middle-old aged patients showed degenerative changes more frequently on both sides. The transition of bone changes to the improved group occurred most commonly in both the adolescent and middle-old aged patients. The adolescent patients were more likely to improve than middle-old aged patients. In the adolescent patients, loss of erosion and subjective symptoms occurred in shorter periods than in the middle-old aged patients. In the adolescent patients, the transition of erosion was distributed into proliferative, normal, and shortening in order. In the middle-old aged patients, the transition of erosion was distributed into shortening, proliferative, and normal in order. Conclusions: The clinical and radiographic features of TMJ OA are a significantly different between the adolescent and middle-old aged patients. Moreover, the difference by age of the adaptive and regenerative capacity of TMJ affects the prognosis of TMJ OA and adolescent patients have a better prognosis after treatment.

키토산이 치주인대 섬유아세포에 미치는 영향 (The effects of chitosan on the human periodontal ligament fibroblasts in vitro)

  • 백정원;이현정;유윤정;조규성;김종관;최성호
    • Journal of Periodontal and Implant Science
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    • 제31권4호
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    • pp.823-832
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    • 2001
  • Periodontal therapy has dealt primarily with attempts at arresting progression of disease, however, more recent techniques have focused on regenerating the periodontal ligament having the capacity to regenerate the periodontium. The effect of chitosan(poly-N-acetyl glucosaminoglycan), a carbohydrate biopolymer extracted from chitin, on periodontal ligament regeneration is of particular interest. The purpose of this study was to evaluate the effect of chitosan on the human periodontal ligament fibroblasts(hPDLFs) in vitro, with special focus on their proliferative properties by M'IT assay, the synthesis of type I collagen by reverse transcription-polymerase chain reaction(RT-PCR) and the activity of alkaline phosphatase(ALP). Fibroblast populations were obtained from individuals with a healthy periodontium and cultured with ${\alpha}MEM$ as the control group. The experimental groups were cultured with chitosan in concentration of 0.01,0.1, 1,2mg/ml. The results are as follows; 1. Chitosan-induced proliferative responses of hPDLFs reached a plateau at the concentration of O.lmg/ml(p<0.05). 2. When hPDLFs were stimulated with 0.lmg/ml chitosan, mRNA expression of type I collagen was up-regulated. 3. When hPDLFs were stimulated with 0.lmg/ml chitosan, ALP activity was significantly up-regulated(p<0.05). In summary, chitosan(0.lmg/ml) enhanced the type I collagen synthesis in the early stage, and afterwards, facilitated differentiation into osteogenic cells. The results of this in vitro experiment suggest that chitosan potentiates the differentiation of osteoprogenitor cells and may facilitate the formation of bone.

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한약재 추출물의 항산화 및 사구체혈관간세포 증식 억제활성 탐색 (Screening of Traditional Medicines for Antioxidative and Anti-proliferative Effects on Rat Mesangial Cells)

  • 손은화;장선아;우한구;구현정;한효상;강세찬
    • 한국자원식물학회지
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    • 제26권5호
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    • pp.652-657
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    • 2013
  • 국내 유통되는 한약재를 이용한 신장질환의 예방 및 치료제 개발을 위한 기초 자료를 제시하고자 경동시장에서 구입한 63종의 한약재 추출물을 이용하여 항산화 활성 및 사구체혈관간 세포의 증식억제능 탐색 활성을 확인하였다. 그 결과 강진향, 고련피, 대풍자, 목향이 RMC 세포의 증식을 50% 이상 억제하는 것으로 나타났고, ORAC와 DPPH assay를 통한 항산화 활성을 확인한 결과 백단향, 백렴이 가장 우수한 ORAC 활성 효능을 보였으며, DPPH 라디칼 소거활성에서는 계혈등, 귀전우, 대풍자, 반대해, 백단향, 백렴이 우수한 효능을 나타냈다. 이 중 대풍자 추출물과 목향 추출물은 항산화 활성과 사구체혈관간세포 증식억제능 모두 뛰어난 효능을 나타냈으나, 목향이 함유하고 있는 aristolochic acid는 임상적으로 신장에 독성을 일으켜 신장질환 치료제에서 제외되는 한약재로 알려져 있다. 따라서, 가장 뛰어난 효능을 보인 대풍자 추출물은 신장질환 치료 및 예방을 위한 한약재 후보물질로서 분획별 항산화 활성과 유효성분 규명의 연구가 요구된다.

2-DDG가 FSa II 종양의 성장속도와 증식 능력, 신진대사에 미치는 영향 ; $^{31}P$-자기공명 분광기와 유세포 분석기를 이용한 연구 (Effects of 2-Deoxy-D-Glucose on Metabolic Status, Proliferative Capacity and Growth Rate of FSall Tumor: Observations made by In Vivo $^{31}P$-Nuclear Magnetic Resonance Spectroscopy and Flow Cytometry)

  • 장혜숙;최은경;조정길;임태환;이대근;이윤;조영주;김곤섭
    • Radiation Oncology Journal
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    • 제9권1호
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    • pp.1-6
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    • 1991
  • 2-ODG가 쥐의 섬유육종(FSall)에 미치는 영향에 대한 연구로 에너지 신진대사는 체내에서의 $^{31}P$-자기공명 분광기를 이용하여 관찰하였고 세포 증식 능력은 유세포 분석기를 사용하여 연구하였다. 성장속도는 10개의 세포를 $C_3Hf/Sed$ 쥐의 발등에 이식한 후 3차원적으로 측정하여 관찰하였다. 2-DDG를 투여한 경우에는 이식후 12일에 복강내로 주사하였다. 이식후 12일의 종양의 평균 크기는 $250mm^3$이었다. Fsall종양의 성장속도는 semilog graph의 기울기와 종양의 doubling time으로 측정하였다. 2-DDG를 투여한 후 성장속도가 감속되었다. 5~12일 사이의 성장속도의 기울기가 0.828, 종양의 Idubling time이 0.84일이고 대조군에서는 13~28일 사이의 기울기가 0.218, doubling time이 3.2일인 반면 2-DOG 투여군에서는 성장속도의 기울기가 0.135이고 doubling time이 5.1일이었다. $^{31}P$-자기공명 분광기를 이용하여 2-DDG의 영향을 분석해 본 결과 2-DDG 투여후 종양증식 속도의 감속과 더불어 phosphornonester (PME)와 inorganic phosphate (Pi)의증가속도가 감소하였다. 이것은 2-DDG투여후 세포의 괴사가 감소하였다는 의미가 있다. 유세포 분석기를 이용하여 종양의 증식 능력을 분석한 결과는 2-DDG 투여후 5-phase와 G2+M phase의 DNA분포가 크게 증가하였다. 이것은 2-DDG투여후 세포가 좀더 방사선에 민감한 cycle로 진행함을 의미하는 것으로 해석할 수 있다. 이에 저자들은 2-DDG가 Fsall 종양세포에 미치는 흥미있는 결과를 토대로 방사선 치료에 미치는 영향과실제 이용 가능성에 대하여 더 연구하고자 한다.

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Lentivirus-mediated Silencing of Rhomboid Domain Containing 1 Suppresses Tumor Growth and Induces Apoptosis in Hepatoma HepG2 Cells

  • Liu, Xue-Ni;Tang, Zheng-Hao;Zhang, Yi;Pan, Qing-Chun;Chen, Xiao-Hua;Yu, Yong-Sheng;Zang, Guo-Qing
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권1호
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    • pp.5-9
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    • 2013
  • Rhomboids were identified as the first intramembrane serine proteases about 10 years ago. Since then, the study of the rhomboid protease family has blossomed. Rhomboid domain containing 1 (RHBDD1), highly-expressed in human testis, contains a rhomboid domain with unknown function. In the present study, we tested the hypothesis that RHBDD1 was associated with proliferation and apoptosis in hepatocellular carcinoma using recombinant lentivirus-mediated silencing of RHBDD1 in HepG2 cells. Our results showed that down-regulation of RHBDD1 mRNA levels markedly suppressed proliferation and colony formation capacity of HepG2 human hepatoma cancer cells in vitro, and induced cell cycle arrest. We also found that RHBDD1 silencing could obviously trigger HepG2 cell apoptosis. In summary, it was demonstrated that RHBDD1 might be a positive regulator for proliferative and apoptotic characteristics of hepatocellular carcinoma.