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cDNA array를 이용한 피골세포 분화관련 유전자의 탐색
조영준,이장희,이창섭,이상호 大韓小兒齒科學會 2002 大韓小兒齒科學會誌 Vol.29 No.2
RAW 264.7과 이것이 분화한 파골세포양 세포에서 파골세포 분화관련유전자의 전체적인 유전자 발현을 조사하기 위해 cDNA array 방법을 사용하였다. 1176 cDNA spot grid가 있는 Mouse Atlas cDNA array 결과를 확인하기 위하여 역전사 효소 중합반응 검사를 시행하였다. cDNA array 결과 6개의 유전자가 2.5% 이상 발현이 증가하였으며(PKC beta Ⅱ, POMC, PTEN 등), 16개의 유전자가 2.5%이상 발현이 감소하였다(Osteopontin, Cyclin D1, Cathepsin C, PTMA 등). PKC beta Ⅱ유전자의 역전사-효소 중합 반응 검사 결과 이 유전자를 확인할 수 있었다. 파골세포 분화 결과 RAW 264.7 세포주에 비해 파골세포양 세포에서 PKC beta Ⅱ 유전자가 발현이 많았다. 파골세포 분화 관련 유전자는 RAW 264.7 세포주의 파골세포 분화와 연관을 보였다. To examine the global gene expression of osteoclastogenesis-related genes in RAW 264.7 and its differentiated OCLs through the use of Atlas Mouse cDNA Array 2.1 membranes printed with 1176 well-characterized mouse genes involved in biology. Both samples were screened in parallel using cDNA expression arrays. The array re-sults were additionally validated using RT-PCR. The results of cDNA arrays showed that 6 genes were up-regu-lated >2.5-fold (PKC beta Ⅱ, POMC, PTEN, etc) and 16 genes were down-regulated >2.5-fold (Osteopontin, Cyclin D1, Cathepsin C, PTMA, etc) in both samples at the mRNA level. RT-PCR analysis of PKC beta Ⅱ of these differentially expressed genes gave result consistent with cDNA array findings. The result of osteoclastoge-nesis showed that the PKC beta Ⅱ gene was overexpressed in OCLs compared with RAW264.7 cell line. Osteoclastogenesis-related genes are differentially expressed in RAW264.7 cell line and its differentiated OCLs. its gene overexpression correlates with osteoclast differentiation in RAW264.7 cell line.
강한승,이채관,문덕환,강성구 한국발생생물학회 2003 발생과 생식 Vol.7 No.1
본 연구는 생식주기 중의 흰쥐 자궁에서 발현되는 유전자들 중 adhesion수용체 유전자들의 발현이 프로게스테론(P)에 의하여 차별적으로 조절되는 실험을 하였다. 첫째 실험군은 난소절제 흰쥐와 배란기 흰쥐를 사용하였고(OVX/estrus), 둘째 실험군은 난소절제 흰쥐와 난소절제 후 P4를 주사한 흰쥐를 사용하였다(OVX/OVX+P). 적출한 자궁조직에서 total RNA를 추출, [P]-dATP로 probe를 제작한 후 Rat Atlas away 1. This report aimed at investigating the differential gene expressions of the adhesion receptors between ovariectomized (OVX) and estrus stage rat uteri (OVX vs. estrus pair) using the cDNA expression away analysis. In addition, this report aimed at confirming of the differential gene expressions of the adhesion receptors between OVX and progesterone (P) injected OVX rat uteri (OVX vs. OVX+P pair). RNA samples were extracted from the uterus and reverse-transcribed in the presence of [ P]-dATP. Membrane sets of Rat Atlas array 1.2 II (Clontech) were hybridized with CDNA probe sets. RT-PCR was employed to validate the relative gene expression patterns obtained by the cDNA array. The results were well consistent to cDNA array analysis data except the fold changes of gene expression. Among a total of 1176 cDNAs, 5 genes of adhesion receotor including embigin protein, activated leukocyte cell adhesion molecule, afadin, neuroligin 2, semaphorin Z showed significant (more than 2-fold) changes in the OVX vs. late estrus pair. All of these genes were up regulated in estrus stage than OVX rat uterus. In the OVX vs. OVX+P pair, 4 genes including osteonectin, afadin, neuroligin 2, semaphorin Z showed significant changes. All of these genes were also up regulated in OVX+P injected rat uterus than OVX control. Three genes including afadin, neuroligin 2, semaphorin Z which were up regulated in estrus and OVX+P injected rat uteri of both experimental pairs than OVX rat uteri. These genes seem to be under the control of P.
cDNA expression array를 이용한 전자간증 태반의 Apoptosis 및 매개인자에 대한 연구
최원준 ( Won Jun Choi ),최완성 ( Wan Sung Choi ),이종학 ( Jong Hak Lee ),백원영 ( Won Young Paik ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.2
Objective : The present study was designed to examine whether preeclampsia is associated with an increase in placental apoptosis and to detect differential expression of mediators involved in apoptosis. Methods : Placental tissues from 10 cases of preecla
Dehydroepiandrosterone으로 유도된 흰쥐의 다낭성 난소에서 cDNA array를 이용한 유전자 발현 분석
정철회 ( Chul Hoi Jeong ),김현찬 ( Hyun Chan Kim ),강성구 ( Sung Goo Kang ),김주란 ( Ju Ran Kim ) 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.10
Objective : Polycystic Ovarian Syndrome (PCOS) is the most common endocrine disorder. Chronic anovulation, hyperandrogenism, hirsutism, obesity, infertility and polycystic ovaries (PCO) are clinical hallmarks of PCOS. PCO can be induced in prepubertal rat
Profiling of Gene Expression in Human Keratinocyte Cell Line Exposed to Quantum Dot Nanoparticles
Kim, In-Kyoung,Lee, Seung-Ho,Kim, Yu-Ri,Seo, Sang-Hui,Jeong, Sang-Hoon,Son, Sang-Wook,Kim, Meyoung-Kon The Korean Society of Toxicogenomics and Toxicopro 2009 Molecular & cellular toxicology Vol.5 No.1
Quantum Dot (QD) nanoparticles are used in various industrial applications, such as diagnostic, drug delivery, and imaging agents of biomedicine. Although QDs are extensively used in many medical science, several studies have been demonstrated the potential toxicity of nanoparticles. The first objective of this study was to investigate the nanotoxicity of QDs in the HaCaT human keratinocyte cell line by focusing on gene expression pattern. In order to evaluate the effect of QDs on gene expression profile in HaCaT cells, we analyzed the differential genes which related to oxidative stress and antioxidant defense mechanisms by using human cDNA microarray and PCR array. A human cDNA microarray was clone set, which was sorted for a list of genes correlated with cell mechanisms. We tried to confirm results of cDNA microarray by using PCR array, which is pathway-focused gene expression profiling technology using Real-Time PCR. Although we could not find the exactly same genes in both methods, we have screened the effects of QDs on global gene expression profiles in human skin cells. In addition, our results show that QD treatment somehow regulates cellular pathways of oxidative stress and antioxidant defense mechanisms. Therefore, we suggest that this study can enlarge our knowledge of the transcriptional profile and identify new candidate biomarker genes to evaluate the toxicity of nanotoxicology.
Molecular Characterization of Small Heat Shock Protein(hsp20.8A) from the Silkworm, Bombyx mori
황재삼,구태원,성수일,윤은영,고현정,안미영,김성률,박관호,전재필,강석우,김익수 한국잠사학회 2007 International Journal of Industrial Entomology Vol.15 No.1
To define the molecular mechanism of initiation and termination of diapause during the embryogenesis of silkworms, Bombyx mori, mRNA transcripts from diapausing eggs and diapause activated eggs were compared with differential expression using cDNA array. Among those clones, mRNA transcript from hsp 20.8A, which was expressed at a high level in diapausing eggs that had been incubated at 25oC for 30 days after oviposition, whereas, in the eggs exposed to 15oC for 30 days, 5oC for 60 days, the expression of mRNA decreased. On the other hand, the expression of mRNA during embryogenesis observed abundantly at 4 to 6 day after heat–HCl treatment and later at 9 to 10 day after just before hatching. This result was suggested for us that hsp20.8A was expressed in response to embryogenesis as well as physical stress.
Molecular Characterization of Small Heat Shock Protein(hsp20.8A) from the Silkworm, Bombyx mori
( Jae Sam Hwang ),( Hyun Jeong Go ),( Tae Won Goo ),( Su Il Seong ),( Eun Young Yun ),( Mi Young Ahn ),( Seong Ryul Kim ),( Kwan Ho Park ),( Ik Soo Kim ),( Jae Pil Jeon ),( Seok Woo Kang ) 한국잠사학회 2007 International Journal of Industrial Entomology Vol.15 No.1
To define the molecular mechanism of initiation and termination of diapause during the embryogenesis of silkworm, Bombyx mori, mRNA transcripts from diapausing eggs and diapause activated eggs were compared with differential expression using cDNA array. Among those clones, mRNA transcript from hsp 20.8A, which was expressed at a high level in diapausing eggs that had been incubated at 25°C for 30 days after oviposition, whereas, in the eggs exposed to 15°C for 30 days, 5°C for 60 days, the expression of mRNA decreased. On the other hand, the expression of mRNA during embryogenesis observed abundantly at 4 to 6 days after heat-HCI treatment and later at 9 to 10 days after just before hatching. This result was suggested for us that hsp2o.8A was expressed in response to embryogenesis as well as physical stress.
Molecular Characterization of Small Heat Shock Protein(hsp20.8A) from the Silkworm, Bombyx mori
Hwang, Jae-Sam,Go, Hyun-Jeong,Goo, Tae-Won,Seong, Su-Il,Yun, Eun-Young,Ahn, Mi-Young,Kim, Seong-Ryul,Park, Kwan-Ho,Kim, Ik-Soo,Jeon, Jae-Pil,Kang, Seok-Woo Korean Society of Sericultural Science 2007 International Journal of Industrial Entomology Vol.15 No.1
To define the molecular mechanism of initiation and termination of diapause during the embryogenesis of silkworm, Bombyx mori, mRNA transcripts from diapausing eggs and diapause activated eggs were compared with differential expression using cDNA array. Among those clones, mRNA transcript from hsp20.8A, which was expressed at a high level in diapausing eggs that had been incubated at $25^{\circ}C$ for 30 days after oviposition, whereas, in the eggs exposed to $15^{\circ}C$ for 30 days, $5^{\circ}C$ for 60 days, the expression of mRNA decreased. On the other hand, the expression of mRNA during embryogenesis observed abundantly at 4 to 6 days after heat-HCl treatment and later at 9 to 10 days after just before hatching. This result was suggested for us that hsp20.8A was expressed in response to embryogenesis as well as physical stress.
The Anti-proliferative Gene TIS21 Is Involved in Osteoclast Differentiation
Lee, Soo-Woong,Kwak, Han-Bok,Lee, Hong-Chan,Lee, Seung-Ku,Kim, Hong-Hee,Lee, Zang-Hee 생화학분자생물학회 2002 Journal of biochemistry and molecular biology Vol.35 No.6
The remodeling process of bone is accompanied by complex changes in the expression levels of various genes. Several approaches have been employed to detect differentially-expressed genes in regard to osteoclast differentiation. In order to identify the genes that are involved in osteoclast differentiation, we used a cDNA-array-nylon membrane. Among 1,200 genes that showed ameasurable signal, 19 genes were chosen for further study. Eleven genes were up-regulated; eight genes were down-regulated. TIS21 was one of the up-regulated genes which were highly expressed in mature osteoclasts. To verify the cDNA microarray results, we carried out RT-PCR and real-time RT-PCR for the TIS21 gene. The TIS21 mRNA level was higher in differentiated-osteoclasts when compared to undifferentiated bone-marrow macrophages. Furthermore, the treatment with $1\;{\mu}M$ of a TIS21 antisense oligonucleotide reduced the formation of osteoclasts from the bone-marrow-precursor cells by ~30%. These results provide evidence for the potential role of TIS21 in the differentiation of osteoclasts.
The Anti-proliferative Gene TIS21 Is Involved in Osteoclast Differentiation
( Soo Woong Lee ),( Han Bok Kwak ),( Hong Chan Lee ),( Seung Ku Lee ),( Hong Hee Kim ),( Zang Hee Lee ) 생화학분자생물학회 2002 BMB Reports Vol.35 No.6
The remodeling process of bone is accompanied by complex changes in the expression levels of various genes. Several approaches have been employed to detect differentially-expressed genes in regard to osteoclast differentiation. In order to identify the genes that are involved in osteoclast differentiation, we used a cDNA-array-nylon membrane. Among 1,200 genes that showed a measurable signal, 19 genes were chosen for further study. Eleven genes were up-regulated; eight genes were down-regulated. TIS21 was one of the up-regulated genes which were highly expressed in mature osteoclasts. To verify the cDNA microarray results, we carried out RT-PCR and real-time RT-PCR for the TIS21 gene. The TIS21 mRNA level was higher in differentiated-osteoclasts when compared to undifferentiated bone-marrow macrophages. Furthermore, the treatment with 1 μM of a TIS21 antisense oligonucleotide reduced the formation of osteoclasts from the bone-marrow-precursor cells by ∼30%. These results provide evidence for the potential role of TIS21 in the differentiation of osteoclasts.