• Title, Summary, Keyword: bromodeoxyuridine

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Effects of 5-Aza-2'-Deoxycytidine, Bromodeoxyuridine, Interferons and Hydrogen Peroxide on Cellular Senescence in Cholangiocarcinoma Cells

  • Moolmuang, Benchamart;Singhirunnusorn, Pattama;Ruchirawat, Mathuros
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.957-963
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    • 2016
  • Cellular senescence, a barrier to tumorigenesis, controls aberrant proliferation of cells. We here aimed to investigate cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines using five different inducing agents: 5-aza-2'deoxycytidine, bromodeoxyuridine, interferons ($IFN{\beta}$ and $IFN{\gamma}$), and hydrogen peroxide. We analyzed senescence characteristics, colony formation ability, expression of genes involved in cell cycling and interferon signaling pathways, and protein levels. Treatment with all five agents decreased cell proliferation and induced cellular senescence in immortalized cholangiocyte and cholangiocarcinoma cell lines with different degrees of growth-inhibitory effects depending on cell type and origin. Bromodeoxyuridine gave the strongest stimulus to inhibit growth and induce senescence in most cell lines tested. Expression of p21 and interferon related genes was upregulated in most conditions. The fact that bromodeoxyuridine had the strongest effects on growth inhibition and senescence induction implies that senescence in cholangiocarcinoma cells is likely controlled by DNA damage response pathways relating to the p53/p21 signaling. In addition, interferon signaling pathways may partly regulate this mechanism in cholangiocarcinoma cells.

Distributions of proliferative epithelial cells in gastrointestinal tracts by anti-bromodeoxyuridine monoclonal antibody (Anti-bormodeoxyuridine monoclonal antibody를 이용한 랫드 위(胃)와 장(腸)의 분열 상피세포의 분포에 대하여)

  • Kwak, Soo-dong;Park, Sung-shik;Kang, Won-hwa
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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    • pp.597-603
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    • 1993
  • The purpose of this stady was to investigate division cells by in vivo bromodeoxyuridine(Brdur) immunohistochemistry for labeling the proliferative epithelial cells in the gastrointestinal tracts of rats. Rats were administrated intraperitonially by twice consecutive injections of 24 hr interval with Brdur(0.05mg/g BW/time) and then were sacrificied at 1 hour after last injection. The specimens were taken from the stomach, small intestine(ileum), and large intestine(colon). The well-oriented crypts and villi in the preparations were examined, The crypt columns and villi were devided into 10 segments from crypt base to surface of the lumen or to villis top. Labeling index(LI) was measured by counting the number of Brdur-positive cells against the total number of crypt column cells in the stomach and large intestine and also against the total numbers of crypt column and it's villi epiterial cells in the small intestine. 1. In the stomach, the LI in each part from segment 1 to segment 10 of the crypt column were 4.2%, 5.0%. 6.6%, 9.0%, 11.3%, 15.3%, 9.3%, 15.6%, 11.3%, 0%, respectively and it's mean LI were 8.7%. The Brdur-positive epithelial cells were predominantly located in the middle regions and middle-upper regions of the crypt columns. 2. In the small intestine, the LI in each part from segment 1 to segment 10 of were 62.4%, 50.9%, 27.8%, 22.5%, 18.6%, 12.1%, 7.5%, 4.3%, 2.5%, 1.4%, respectively and it's mean LI were 21.0%. The Brdur-positive epithelial cells were predominantly located in the lower regions of the crypt columns and tended to be less in the higher regions of the villi than that in the crypt column. 3. In the large intestine, the LI in each part from segment 1 to segment 10 of the crypt column were 19.4%, 29.9%, 34.1%, 41.6%, 41.2%, 32.4%, 25.4%, 15.4%, 10.8%, 1.2%, respectively and it's mean LI were 25.1%, The Brdur-positive epithelial cells were predominantly in the middle and middle-lower regions of the crypt columns. 4. The organs with higher LI were ordered as the large intestine(25.1%), small intestine(21.0%) and stomach(8.7%).

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An immunohistochemical study on distribution of proliferating cells in uterus and ovary of progesterone-treated rats (Progesterone이 rat 자궁과 난소의 증식세포 분포에 미치는 영향에 대한 면역조직화학적 연구)

  • Park, Sung-sik;Kwak, Soo-dong
    • Korean Journal of Veterinary Research
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    • v.35 no.2
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    • pp.217-228
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    • 1995
  • The study was designed to investigate the effects of progesterone on the reproductive system. This investigation was performed by immunohistochemical methods using anti-bromodeoxyuridine-antibody following bromodeoxyuridine(Brdur) injection for labeling proliferating cells in the uterus and ovary of rats. Sixteen female rats(Wistar), weighing initially 300g, were randomly allotted into ovariectomized and unovariectomized large groups. These two large groups were subdivided into three subgroups of control, 3-day and 6-day groups, respectively. 3-days and 6-days group were injected with 1mg of progesterone/rat/day for 3 or 6 days, respectively. In gross findings, the uterus of ovariectomized groups markedly atrophied, and were not hypertrophied by progesterone injection for 3 days or 6 days and the uterus of unovariectomized groups also were not hypertrophied. Labeling index(LI, %) was measured by counting the number of Brdur-positive cells from 300 to 3,000 cells per layer in the uterus tissue. The average LI of the uterus in unovariectomized groups was higher than that of ovariectomized groups. The subgroups with higher LI in unovariectomized groups were ordered as 6-day group, 3-day group. So progesterone considerably effected to the proliferating of the cells in the uterus of unovariectomized groups. The layers with higher LI in the uterus wall were ordered as the functional zone of endometrium, epithelial layer of endometrium, basal zone of endometrium, myometrium and perimetrium. The cell types with higher LI in the uterus of unovariectomized groups were ordered as the surface epithelial cells, stromal cells, glandular epithelial cells and muscle cells. Growing follicles with proliferating cells from secondary and tertiary follicles in the ovary of unovariectomized groups appeared to be 37.66% in control group, 39.23% in 3-day groups, 39.47% in 6-day groups. Mature follicles in the ovary were more number in control group than those in 3-day groups but not appeared in 6-day groups. So progesterone not nearly effects to the number of the growing follicles but appeared to be related to suppression of the development and protrusion of the mid-tertiary and mature follicles on the ovary surface. The cell types with higher LI in the ovary of unovariectomized groups were respectively ordered as granulosa cells, theca interns cells in secondary follicles; theca interna cells, granulosa cells, theca externa cells in tertiary follicles; fibroblasts, theca in terns cells in atretic follicles; fibroblasts, luteal cells in corpus luteum.

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EVALUATION OF CELL PROLIFERATION IN EAR AND LYMPH NODE USING BRDU IMMUNOHISTOCHEMISTRY FOR MOUSE EAR SWELLING TEST

  • Lee, Jong-Kwon;Park, Jae-Hyun;Kim, Hyung-Soo;Jung, Seung-Tae
    • Proceedings of the Korean Society of Toxicology Conference
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    • pp.97-97
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    • 2002
  • A mouse ear swelling test (MEST) has been developed as an alternative to guinea pig models for contact sensitization potential. However, the MEST relies on a quantitative measurement of ear swelling as an endpoint by micrometer. In this study, we aimed to investigate the cell proliferation in ear and lymph node using Bromodeoxyuridine (BrdU) immunohistochemistry as a possible reliable marker for MEST.(omitted)

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A Non-radioisotopic Endpoint Using Bromodeoxyuridine ELISA Method for Murine Local Lymph Node Assay (BrdU ELISA를 이용한 국소 림프절 시험법의 비방사선법 연구)

  • 이종권;박재현;박승희;김형수;정승태;엄준호;윤소미;장은정;최광식
    • Toxicological Research
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    • v.19 no.2
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    • pp.133-139
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    • 2003
  • Allergic contact dermatitis may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. However, there is a need to develop a nonradioisotopic endpoint for the LLNA, because of the radioisotopic method's requiring the use of special facilities. In this study, we investigated the development of a nonradioisotopic endpoint for LLNA using ELISA (enzyme-linked immunosorbent assay). Female Balb/c mice were treated by the topical application on the dorsum of both ears with four different strong sensitizers, 2,4-dinitrochlorobenzene (DNCB), oxazolone (OXZ), toluene diisocyanate (TDI), and trimellitic anhydride (TMA), and a strong irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The proliferation of cells in the auricular Iymph node was analyzed by means of the labelling index (Ll) of bromodeoxyuridine (BrdU) incorporation into cells. The weights of the Iymph nodes in the mice treated with allergens, DNCB, OXZ, TDl and TMA were increased compared to the vehicle control. The stimulation index (Sl) of mice treated with DNCB, OXZ, TDl, and TMA was over three-fold increase compared to the vehicle control. However, the S1 of mice exposed to SLS was not significantly increased compared to the vehicle control, while the lymph node weight of SLS was significantly increased. These results suggest that the LLNA modified endpoint using ELISA based on BrdU incorporation could provide a useful method of screening for irritants and allergens.

Immunohistochemical observations of proliferating cells in distal epiphyseal tissue of chicken femurs (닭의 대퇴부 골단조직의 세포증식에 대한 면역조직화학적 관찰)

  • Kwak, Soo-dong;Kim, Chong-sup;Kang, Chung-boo
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.237-242
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    • 1994
  • The present study was focussed to assess the proliferating cells in the distal epiphyseal tissue of the chicken femur by immunohistochemical staining methods. Four chickens were administrated intraperitoneally by twice consecutive injections, 1 day interval with bromodeoxyuridine(Brdur, 0.05 mg/gm BW/time), and then were killed by exsanguination of jugular vein at 2 hours after last injection. Samples were taken from femur distal epiphyseas of chicken. Labeling indexes(LI) were calculated as the ratio of the number of anti-Brdur monoclonal antibody-labeled cells in the each tissue layers from basal layer of the integument to bone marrow. The overall LI were found to be $13.90{\pm}3.44%$, $30.03{\pm}7.52%$, $16.00{\pm}9.41%$, $0.00{\pm}0.00%$ and $60.03{\pm}13.39%$ at basal layer of integument, perichordrium, reseving zone in cartilage, hypertrophic zone in cartilage and bone marrow respectively. LI in proliferating zone of cartilage were found to be $36.99{\pm}7.59%$, $32.83{\pm}5.38%$ and $22.02{\pm}6.27%$ at reserving zone side region, middle region, and hypertrophic zone side region respectively. The tissue layers with higher LI were odered as bone marrow, reserving zone side region in proliferating zone, middle region in proliferating zone, perichondrium, hypertrophic zone side region in proliferating zone. reserving zone of cartilage and basal layer of integument. These data indicate that the overall LI in the each tissue layer of distal epiphyseas of the chicken femur were concluded to be higher than that in another tissue of adult birds but hypertrophic zone of cartlage were appeared to be not proliferating cells.

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AN IMMUNOHISTOCHEMICAL STUDY ON DNA SYNTHESIS OF SALIVARY GLAND TISSUE CEllS AND ENDOTHELIAL CELL AFTER IRRADIATION (방사선조사 후 타액선 세포와 혈관 내피세포의 DNA합성에 관한 면역조직학적 연구)

  • Shin Jong-Sup;You Dong-Soo
    • The Journal of Korean Academy of Maxillofacial Radiology
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    • v.21 no.2
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    • pp.183-197
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    • 1991
  • After single fraction of 2, 5, 10 Gy irradiation on submandibular gland of 40 male rats, weighing 150gm, respectively, these animal were sacrificed two hours after 0.1㎎/g bromodeoxyuridine (Sigma) peritoneal injection in 1, 3, 7, 15 hours, 1, 3, 7 days after irradiation. And excised submandibular gland were fixed in Carnoy's and Bouin's solution for 2 hours. Paraffin sections were stained with H&E, and PAS for the observation of the change of salivary gland tissue, and with Feulgen for the study of the DNA distribution, and immunohistochemically stained with anti-bromodeoxyuridine (Sanbyo Co.) for detection of DNA synthetic cells in order to study the distribution of DNA synthetic cells of salivary gland tissue and endothelium after irradiation in 5 different sites of 6 slides on X 200 high power field. The results were as followings. 1. In PAS staining 3 days after 5Gy irradiation, decreased mucine secretion of serous cells were found, and 7 days after l0Gy irradiation, decreased mucine secretion of mucous cells were found. 2. In histopathologic features, degeneration of serous cells were found in 3 days after 2 Gy irradiation and there was little change in mucous cells and excretory duct cells. 3. In Feugen staining, 3 days after 2 Gy, 5 Gy irradiation, more high percentage of DNA synthetic cells were found in intercalated duct cells, striated duct cells and excretory duct cells than in BrdU staining. 4. In immunohistochemical features, DNA synethsis of serous cells and granular convoluted tubular cells abruptly decreased in early period after irradiation and showed no recovery in 7 days after irradiation but there was an increase in DNA synthesis of intercalated duct cells, striated duct cells and excretory duct cells, which have less S-phase cells comparatively, in 7 days after 2 Gy, 5 Gy irradiation. 5. In immunohistochemical features, the DNA synthesis of endothelial cells was continuously decreased after irradiation but showed slight increase in 7 days after 2 Gy and S Gy irradiation.

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Turnover of biliaiy epithelial cells in Clonorchis sinensis infected rats (간흡충에 감염된 흰쥐 담관 상피세포의 증식 양상)

  • 홍성태;고원규
    • The Korean Journal of Parasitology
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    • v.31 no.2
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    • pp.83-90
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    • 1993
  • We performed bromodeoxyuridine (BrdU) staining to observe the proliferation pattern of epithelial cells on the biliaJy mucosa in Clonorchis sinensis infection. Albino rats were infected with 100 metacercariae each and their livers were processed for histopathological observation after BrdU injection. Five to six sites in the liver of a rat were selected for paraffin section, and stained immunohistochemically to visualize BrdU incorporating cells. The flukes were mainly in the common bile duct and right or left hepatic bile ducts. The proportion of stained epithelial cells in the infected bile ducts where the worms were found on the section was 2.9-10.2% at 1 week after infection. 7.3-12.8% at 2 weeks, 7.3-13.4% at 5 weeks, and 8.4-14.8% at 15 weeks while in the non-infected ducts o to 2.7% cells were stained. The stained cells were mainly at the base of the mucosal layer. It is suggested that mucosal epithelial cells of the bile ducts infected with C. sinensis become hyperplastic mainly by direct and local stimulation of the worms.

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Acupuncture Stimulation to HT8 Enhances Cell Proliferation in Hippocampus on an Epilepsy Mouse Model (마우스 간질 동물모델에서 소부혈 자침이 해마 치상회의 신경세포증식에 미치는 영향)

  • Kim, Seung-Tae;Park, Hae-Jeong;Hong, Mee-Sook;Kim, Seung-Nam;Doo, Ah-Reum;Yin, Chang-Shik;Lee, Hye-Jung;Chung, Joo-Ho;Park, Hi-Joon
    • Korean Journal of Acupuncture
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    • v.27 no.2
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    • pp.49-56
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    • 2010
  • 목적 : 뇌의 신경세포 증식은 해마 치상회와 뇌실하영역에서만 나타나는 현상이다. Kainic acid(KA)를 이용한 간질 동물모델을 연구하던 중 침이 해마 치상회의 신경세포증식을 촉진하는 현상을 발견하여 이를 보고하고자 한다. 방법 : 수컷 ICR계 생쥐를 Saline(n=8), KA(n=8), KA+Acu(n=8)의 세 군으로 나누고, 모든 생쥐들에게 KA 주입 3일 전부터 1일 1회씩 5'-bromodeoxyuridine(BrdU)을 3일간 주입하였다. Saline군에는 멸균된 생리식염수를 뇌실 내에 주입하였고, KA군 및 KA+Acu군에는 $0.1{\mu}g$의 KA를 뇌실 내에 주입하였으며, KA+Acu군에 속한 쥐들에게는 KA 주입 2일전, 1일전, 주입 직후에 양쪽 소부(少府)(HT8)에 자침하였다. KA 주입 3시간 후 쥐의 뇌를 적출하고 해마 치상회부위의 BrdU 및 neuropeptide Y (NPY)의 발현을 측정하였다. 결과 : 소부(少府) 자침이 KA의 독성으로 인한 신경세포의 파괴를 줄여주었으며, BrdU 양성 세포 및 NPY를 유의하게 증가시켰다. KA 주입시 세포증식이 일어나긴 하나, 3시간 안에는 거의 일어나지 않는다. 결론 : 소부(少府) 자침이 해마 치상회의 신경세포증식을 촉진하며, 이는 KA의 효과가 아닌 KA 투여 전 소부(少府) 자침으로 인한 것으로 사료된다.

Stress-induced Decrease of Granule Cell Proliferation in Adult Rat Hippocampus: Assessment of Granule Cell Proliferation Using High Doses of Bromodeoxyuridine Before and After Restraint Stress

  • Kim, Sung-Jin;Lee, Kuem-Ju;Shin, You-Chan;Choi, Song-hyen;Do, Eunju;Kim, Sangduk;Chun, Boe-Gwun;Lee, Min-Soo;Shin, Kyung-Ho
    • Molecules and Cells
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    • v.19 no.1
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    • pp.74-80
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    • 2005
  • Stress is known to inhibit granule cell proliferation in the hippocampus. However, recent studies suggest that the commonly used dose of bromodeoxyuridine (BrdU) is insufficient to label all fractions of granule cells. Furthermore, stress-induced changes in BrdU availability may influence the labeling of newly born cells. To investigate whether changes in BrdU availability affect measurements of stress-induced granule cell proliferation, granule cell proliferation was assessed using injection of high doses of BrdU before and after restraint stress lasting 1 h. In addition, to determine whether stress-induced changes in plasma corticosterone levels were influenced by the BrdU, time-dependent changes in plasma corticosterone levels over 2 h after BrdU injection were compared with total accumulated plasma corticosterone levels [as determined by areas under the curve (AUC)]. Restraint stress significantly reduced the numbers of BrdU-labeled cells and clusters in the granule cell layer (GCL) of rats that received BrdU after stress, and decreases of similar magnitude were observed when the rats were given BrdU before stress. BrdU injection enhanced the stress-induced plasma corticosterone response, but there was no difference between the mean AUCs of plasma corticosterone levels of animals injected with BrdU before or after stress. These observations suggest that restraint stress decreases granule cell proliferation, and that this may be influenced by the extent and duration of plasma corticosterone increases rather than by changes in the availability of BrdU.