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1.
Rio de Janeiro; s.n; 2018. 58 p. ilus.
Thesis in Portuguese | LILACS | ID: biblio-1562389

ABSTRACT

Os erros no padrão de imprinting são caracterizados como alterações epigenéticas por não afetarem a sequência nucleotídica do DNA mas modificarem a expressão gênica. Diversos aspectos do desenvolvimento embrionário, incluindo ocorrência de malformações congênitas e síndromes diversas, como a síndrome de Beckwith-Wiedemann (SBW; OMIM#130650) ocorrem em decorrência de eventos epigenéticos. A etiologia molecular de SBW é descrita com base em alterações epigenéticas na região cromossômica 11p15.5, que possui genes cuja expressão é regulada por dois centros de imprinting (IC) ou regiões diferencialmente metiladas (DMR). O IC1, que controla a expressão dos genes IGF2 e H19 apresenta metilação no alelo paterno e ausência de metilação no alelo materno; o IC2 encontra-se metilado normalmente no alelo materno, participando no controle da expressão dos genes CDKN1C, KCNQ1 e KNCQ1OT1. A etiologia de SBW também está associada a mutações pontuais em genes como CDKN1C, que codifica um inibidor de kinase dependente de ciclina da fase G1 do ciclo celular. O objetivo deste trabalho é investigar alterações genéticas e epigenéticas em amostras de DNA de 21 pacientes com SBW oriundos do Ambulatórido de Genética Médica do Instituto Nacional Fernandes Figueira IFF/FIOCRUZ. A abordagem metodológica consistiu na análise do perfil de metilação dos dois centros de imprinting de 11p15.5 com uso da ténica de MS-MLPA; análise de microssatélites dessa região, visando discriminar alterações epigenéticas em ambos os centros de imprinting de possíveis dissomias uniparentais (UPD); e a amplificação do gene CDKN1C, para sequenciamento de Sanger. Dos 21 pacientes analisados inicialmente por MS-MLPA, sete não apresentaram nenhuma alteração epigenética. Após reavaliação clínica foi confirmado o diagnóstico de SBW em três desses sete pacientes enquanto que os outros quatro pacientes apresentaram características clínicas isoladas associadas à SBW, sendo excluídos da amostra. Nos 14 pacientes restantes a análise de MS-MLPA identificou 10 casos de perda isolada de metilação no IC2, três casos de deleção em IC2 e um caso de ganho de metilação em IC1 com perda em IC2 (UPDp). Este trabalho confirmou estudos anteriores sobre a ocorrência de alterações epignéticas nesta síndrome; em consequência, permitindo uma orientação clínica e antecipação do monitoramento de pacientes quanto ao risco de tumores pediátricos.


Errors in imprinting pattern are characterized as epigenetic alterations, not affecting the nucleotide sequencing on DNA but affecting the gene expression. Several aspects of embrionary development, including occurrence of abnormalities and congenital disorders, as Beckwith-Wiedemann syndrome (BWS; OMIM#130650) occur due epignetics events. The molecular etiology of SBW is described based on epigenetic alterations in 11p15.5 chromossomic region, wich includes genes that have the expression regulated by two imprinting centers (IC) or differentialy methylated region (DMR). The IC1 regulates the gene expression of IGF2 and H19 genes. This IC shows a high methylation level only in the paternal allele. The IC2, normally methylated on the maternal allele, controls de gene expression of CDKN1C, KCNQ1 e KNCQ1OT1 genes. Besides this alterations, SBW's etiology is also associated with pontual mutations in CDKN1C gene, which encondes a cyclin dependent kinase inhibitor, acting on G1 celular cycle phase. The purpose of this study is to investigate genetics and epigenetics alterations in DNA samples of 21 patients with BWS from Instituto Nacional Fernandes Figueira IFF/FIOCRUZ. The methodological approach was based on the methylation profile's analysis of the two 11p15.5 imprinting centers using the MS-MLPA technique; microsatellite analysis of this region, aiming to distinguish epigenetic alterations on both imprinting centers from possible uniparental disomy (UPD); and CDKN1C gene Sanger sequencing. Seven of 21 patients analysed by MS-MLPA did not shown any epigenetic alteration. After a clinical revaluation the SBW diagnosys was confirmed to three of this sevens patients while the reamining four only shown isolated clinial features associated with BWS. On 14 patients MS-MLPA analysis identified 10 cases of isolated loss of methylation on IC2, three cases of IC2 deletions and one case with both gain of methylation on IC1 and loss of methylation on IC2 (UDPp). This study confirmed previous researches about the epigenetic alteration occurence ih BWS; in consequence, allowing a clinical monitoring to this patients due the risk of pediatric tumors.


Subject(s)
Humans , Beckwith-Wiedemann Syndrome/diagnosis , Beckwith-Wiedemann Syndrome/genetics , Gene Expression , Epigenesis, Genetic , Cyclin-Dependent Kinase Inhibitor p57 , Epigenomics , Brazil
2.
Zhongguo yi xue ke xue yuan xue bao ; Zhongguo yi xue ke xue yuan xue bao;(6): 253-258, 2015.
Article in English | WPRIM | ID: wpr-257650

ABSTRACT

<p><b>OBJECTIVE</b>To explore the functions of miR-9 and miR-9(*) in SAMP8 mice during the aging and their possible mechanisms.</p><p><b>METHODS</b>SAMP8 mice(4-,8-,12-month old,respectively)were selected,three age-matched SAMR1 mice were used as the control group with three mice in each group. The brains were collected and then sectioned for in situ hybridization of miR-9 and miR-9(*). Mimics or inhibitors of miR-9 and miR-9(*) were transfected into N2a cells,and the effects of overexpression or knockdown of the microRNAs on the cell cycle were detected by flow cytometry. Target genes were predicted by bioinformatic analysis and confirmed by dual luciferase assay.</p><p><b>RESULTS</b>Expressions of miR-9 and miR-9(*) in hippocampus of SAMP8 mice were lower than those of SAMR1 mice. Knockdown of miR-9 and miR-9(*) induced a prolonged G1 phase and a shortened S phase in N2a cells;in contrast,miR-9 and miR-9(*) overexpression showed opposite effects. The predicted target genes of miR-9 were PSEN1,SCN2B,MAP3K3,and BACE1,and that of miR-9(*) was CDKn1c. Dual luciferase reporter gene assay showed that miR-9 targeted MAP3K3 while miR-9(*) targeted CDKn1c.</p><p><b>CONCLUSION</b>miR-9 and miR-9(*) play an important role during aging via the target genes MAP3K3 and CDKn1c in the SAMP8 mice.</p>


Subject(s)
Animals , Mice , Aging , Brain , Cyclin-Dependent Kinase Inhibitor p57 , MicroRNAs
3.
Zhonghua Bing Li Xue Za Zhi ; (12): 868-873, 2015.
Article in Chinese | WPRIM | ID: wpr-278510

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical utility of short tandem repeats(STR) genotyping technique for diagnosis of partial hydatidiform moles (PHM).</p><p><b>METHODS</b>Ten cases with the original diagnosis of PHM and six cases diagnosed as "favour PHM" or "abnormal villous, PHM not excluded" were selected for the study. The clinical information and follow-up data were reviewed. Histopathologic features were evaluated along with p57 immunohistochemistry. After DNA extraction from each sample, genotyping was performed by AmpFlSTR(®) Identifiler™ PCR kit to amplify 15 STR polymorphism loci plus the amelogenin gender-determining in a single robust PCR.</p><p><b>RESULTS</b>The age of patients ranged from 18 to 49 years (mean=29 years, median=29 years). Two villous populations (7/16), irregular villous contour (13/16), at least moderate trophoblastic hyperplasia (2/16), cistern formation (8/16), syncytiotrophoblastic knuckles (14/16), trophoblastic pseudoinclusions (6/16) and nucleated fetal red blood cells (8/16) were presented in these cases. Of the cases in the study, STR genotyping identified 4 monospermic complete hydatidiform moles (MCM), 3 dispermic partial hydatidiform moles (DPM) and 9 hydropic abortions (HA). The misdiagnosis rate was 13/16 only relied on morphology evaluation. Immunostaining of p57 showed 3/4 of MCM were focally positive (<5%-20%+), 1/4 of MCM were diffusely positive (70%+), 3/3 of DPM were diffusely positive (≥50%+), 7/9 of HA were diffusely positive (≥50%+), and 2/9 of HA were focally positive (10%+).</p><p><b>CONCLUSIONS</b>Combination of histomorphologic evaluation and p57 immunostaining is insufficient for a definitive diagnosis of PHM. STR genotyping offers an accurate diagnosis of PHM.</p>


Subject(s)
Adolescent , Adult , Female , Humans , Middle Aged , Pregnancy , Young Adult , Abortion, Spontaneous , Cyclin-Dependent Kinase Inhibitor p57 , Metabolism , Genotype , Genotyping Techniques , Hydatidiform Mole , Diagnosis , Genetics , Immunohistochemistry , Microsatellite Repeats , Polymerase Chain Reaction , Trophoblasts , Pathology , Uterine Neoplasms , Diagnosis , Genetics
4.
Article in English | WPRIM | ID: wpr-250343

ABSTRACT

Neural stem cells (NSCs) proliferation can be influenced by repetitive transcranial magnetic stimulation (rTMS) in vivo via microRNA-106b-25 cluster, but the underlying mechanisms are poorly understood. This study investigated the involvement of microRNA-106b-25 cluster in the proliferation of NSCs after repetitive magnetic stimulation (rMS) in vitro. NSCs were stimulated by rMS (200/400/600/800/1000 pulses per day, with 10 Hz frequency and 50% maximum machine output) over a 3-day period. NSCs proliferation was detected by using ki-67 and EdU staining. Ki-67, p21, p57, cyclinD1, cyclinE, cyclinA, cdk2, cdk4 proteins and miR-106b, miR-93, miR-25 mRNAs were detected by Western blotting and qRT-PCR, respectively. The results showed that rMS could promote NSCs proliferation in a dose-dependent manner. The proportions of ki-67+ and Edu+ cells in 1000 pulses group were 20.65% and 4.00%, respectively, significantly higher than those in control group (9.25%, 2.05%). The expression levels of miR-106b and miR-93 were significantly upregulated in 600-1000 pulses groups compared with control group (P<0.05 or 0.01 for all). The expression levels of p21 protein were decreased significantly in 800/1000 pulses groups, and those of cyclinD1, cyclinA, cyclinE, cdk2 and cdk4 were obviously increased after rMS as compared with control group (P<0.05 or 0.01 for all). In conclusion, our findings suggested that rMS enhances the NSCs proliferation in vitro in a dose-dependent manner and miR-106b/p21/cdks/cyclins pathway was involved in the process.


Subject(s)
Animals , Rats , Animals, Newborn , Biomarkers , Metabolism , Cell Proliferation , Genetics , Cyclin-Dependent Kinase 2 , Genetics , Metabolism , Cyclin-Dependent Kinase 4 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Metabolism , Cyclins , Genetics , Metabolism , Gene Expression Regulation , Hippocampus , Cell Biology , Metabolism , Ki-67 Antigen , Genetics , Metabolism , Magnetic Fields , MicroRNAs , Genetics , Metabolism , Neural Stem Cells , Cell Biology , Metabolism , Primary Cell Culture , Rats, Sprague-Dawley , Signal Transduction
5.
Article in Chinese | WPRIM | ID: wpr-330959

ABSTRACT

This study was purposed to explore the expression of p57kip2 in the bone marrow of patients with de novo myelodysplastic syndrome (MDS) and its role in MDS pathogenesis, as well as the relationship between the expression of p57kip2 and SDF-1/CXCR4 signal. The expression of p57kip2 and CXCR4 in 67 de novo MDS patients was measured by real-time quantitative PCR. The percentage of CD34(+) cells in the bone marrow from MDS patients was measured by flow cytometry. 18 healthy volunteers were recruited for control. The effect of SDF-1 on p57kip2 expression in bone marrow mononuclear cell (BMMNC) from MDS or normal controls was investigated in vitro, and difference between them was compared. The results showed that low-risk MDS and high-risk MDS displayed a significant reduction of p57kip2 mRNA expression in BMMNC compared with that in control group (P < 0.001) and there was a negative correlation between p57kip2 expression and percentage of CD34(+) (r = -0.458, P < 0.001); the patients with abnormal karyotype showed lower expression of p57kip2 gene, compared to patients with normal karyotype (P = 0.045). Although the expression of CXCR4 had no difference between MDS patients and normal controls, a positive correlation between p57kip2 and CXCR4 in MDS patients was still found (r = 0.609, P < 0.001). Moreover, SDF-1 increased p57kip2 expression in normal BMMNC in dose-dependent manner, but BMMNC from MDS patients showed no response to SDF-1. SDF-1-induced p57 expression was blocked by AMD3100. It is concluded that the low expression of p57 gene in MDS may play a role in the pathogenesis of MDS. Furthermore, SDF-1-induced p57kip2 expression in BMMNC, and the decreasing response of BMMNC to SDF-1 may contribute to the low expression of p57kip2 in MDS patients.


Subject(s)
Humans , Case-Control Studies , Chemokine CXCL12 , Metabolism , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Metabolism , Flow Cytometry , Myelodysplastic Syndromes , Genetics , Metabolism , Receptors, CXCR4 , Metabolism
6.
Article in Chinese | WPRIM | ID: wpr-237128

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of microRNA-221 (miR-221) and CDKN1C/P57 in colorectal carcinoma (CRC) and adjacent non-cancerous tissues. The effect of miR-221-specific inhibitor on cell proliferation and apoptosis in CRC cells was also assessed.</p><p><b>METHODS</b>The expression of miR-221 was detected by real-time RT-PCR. CDKN1C/P57 mRNA and corresponding protein expression pattern were detected by semi-quantitative RT-PCR and Western-blot. The specific 2'-methoxy-modified RNA oligonucleotide of miR-221(miRNA inhibitor,anti-miR-221) was designed, synthesized and transfected into Caco2 cell by liposome. Finally, the status of CRC cell proliferation and apoptosis were detected by MTT assay and flow cytometry.</p><p><b>RESULTS</b>The expression of miR-221 was significantly up-regulated in CRC tissues as compared to the adjacent non-cancerous tissues(2.041±1.401 vs. 0.806±0.341, P<0.01). There was no significant difference in CDKN1C/P57 mRNA expression between CRC and non-cancerous tissues, whereas CDKN1C/P57 protein markedly decreased in CRC (3.019±1.708 vs. 0.972±0.316, P<0.01). miR-221-specific inhibitor significantly enhanced CDKN1C/P57 protein expression, inhibited proliferation of CRC cells and induced apoptosis of CRC cells(P<0.01).</p><p><b>CONCLUSIONS</b>miR-221 inhibits CDKN1C/P57 expression by post-transcriptional gene silencing to promote CRC development and progression. miR-221-specific inhibitor potentially inhibits the growth of CRC cells. Therefore, it may be a new target for the biologic therapy for CRC.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Apoptosis , Genetics , Caco-2 Cells , Cell Line, Tumor , Cell Proliferation , Colorectal Neoplasms , Genetics , Metabolism , Pathology , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Metabolism , MicroRNAs , Genetics , RNA Interference
7.
Article in Chinese | WPRIM | ID: wpr-246911

ABSTRACT

<p><b>OBJECTIVE</b>To study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.</p><p><b>METHODS</b>Immunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.</p><p><b>RESULTS</b>The expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).</p><p><b>CONCLUSIONS</b>The decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.</p>


Subject(s)
Humans , Cicatrix , Metabolism , Pathology , Cicatrix, Hypertrophic , Metabolism , Pathology , Cyclin-Dependent Kinase Inhibitor p57 , Metabolism , Fibroblasts , Metabolism , Serpins , Metabolism
8.
Article in Chinese | WPRIM | ID: wpr-265760

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the regulatory effect of microRNA-221 (MIR221) on CDKN1C/p57 expression in colon carcinoma cells in vitro.</p><p><b>METHODS</b>Caco2 cells were treated with or without anti-p57-siRNA prior to the addition of pre-MIR221 or anti-MIR221. The MIR221 expression pattern was detected by real-time RT-PCR, and the mRNA and protein levels of CDKN1C/p57 expression were detected using semi-quantitative RT-PCR and Western blotting. Caco2 cell proliferation following the treatment was detected with MTT assay. CDKN1C/p57 3'-UTR fragment was amplified by PCR from the genome DNA of human colon and inserted into a luciferase reporter plasmid. The luciferase reporter plasmid construct was then transfected into Caco2 cells along with pre-MIR221 or anti-MIR221, and the luciferase activity in the transfected cells was detected.</p><p><b>RESULTS</b>MIR221-specific inhibitor significantly up-regulated CDKN1C/p57 protein expression in Caco2 cells (P<0.01). Anti-MIR221 could markedly inhibit Caco2 cell proliferation, and the inhibitory effect was obviously abolished by pretreatment with anti-p57-siRNA, suggesting that the inhibition was mediated by CDKN1C/p57 (P<0.01). A significant increase of luciferase activity was detected in Caco2 cells co-transfected with the luciferase reporter plasmid construct and anti-MIR221 (P<0.01).</p><p><b>CONCLUSIONS</b>MIR221 can interact with the target site on the 3'-UTR of CDKN1C/p57 mRNA to inhibit CDKN1C/p57 expression by post-transcriptional gene silencing to promote colon carcinoma cell proliferation, suggesting the value of MIR221 as a potential target for treatment of colon carcinoma.</p>


Subject(s)
Humans , 3' Untranslated Regions , Caco-2 Cells , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Metabolism , Down-Regulation , MicroRNAs , Pharmacology , RNA, Messenger , Genetics , Metabolism
9.
Zhonghua Bing Li Xue Za Zhi ; (12): 694-697, 2011.
Article in Chinese | WPRIM | ID: wpr-358262

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of p57 and p53 immunohistochemistry in the differential diagnosis of hydropic abortion, partial hydatidiform mole and complete hydatidiform mole.</p><p><b>METHODS</b>Immunohistochemical stains (EnVision method) for p57 and p53 were performed in tissue samples of normal placenta chorionic villi (n=10), abortion chorionic villi (n=12), partial hydatidiform (n=23) and complete hydatidiform moles (n=20).</p><p><b>RESULTS</b>The expression of p57 was predominantly localized in the nuclei of villous cytotrophoblasts and stromal cells. The positive rates of p57 in normal placenta, hydropic abortion and partial hydatidiform mole were 10/10, 12/12, and 100% (23/23), respectively, with no significant difference among the groups (P>0.05). However, none of the complete hydatidiform moles analyzed exhibited p57 positivity in cytotrophoblasts and stromal cells. There was a significant difference between partial and complete hydatidiform moles (P<0.05). The expression of p53 was observed in the nuclei of cytotrophoblastic cells and intermediate trophoblasts. No p53 expression was seen in normal placenta and only 1 of 12 hydropic abortion showed p53 positivity. The positive rates of p53 expression in partial and complete hydatidiform mole were 60.9% (14/23) and 85.0% (17/20) respectively. It was significantly higher in partial hydatidiform mole than that in hydropic abortion. A significant difference was also found between partial and complete hydatidiform moles (P<0.05).</p><p><b>CONCLUSIONS</b>Our findings confirm that p57 immunohistochemistry assists the differential diagnosis of complete hydatidiform mole from partial hydatidiform mole. Expression of p53 may be helpful in distinguishing partial hydatidiform mole from hydropic abortion.</p>


Subject(s)
Female , Humans , Pregnancy , Abortion, Spontaneous , Diagnosis , Metabolism , Pathology , Cyclin-Dependent Kinase Inhibitor p57 , Metabolism , Diagnosis, Differential , Hydatidiform Mole , Diagnosis , Metabolism , Pathology , Immunohistochemistry , Stromal Cells , Metabolism , Trophoblasts , Metabolism , Tumor Suppressor Protein p53 , Metabolism , Uterine Neoplasms , Diagnosis , Metabolism , Pathology
10.
Zhonghua Bing Li Xue Za Zhi ; (12): 580-584, 2009.
Article in Chinese | WPRIM | ID: wpr-249035

ABSTRACT

<p><b>OBJECTIVE</b>To study the value of combined use of paternally imprinted gene product p57(KIP2) immunohistochemistry and flow cytometry in the differential diagnosis of placental hydropic diseases.</p><p><b>METHODS</b>A total of 32 cases of hydropic placenta with DNA polymorphism information were collected, and the genetic results were used as basis for the diagnosis of complete hydatidiform moles (CHM), partial hydatidiform moles (PHM) or hydropic abortions. All cases were examined by histology, p57(KIP2) immunohistochemical staining (EnVision method) and flow cytometry DNA ploidy analysis. The p57(KIP2) immunohistochemical staining and DNA ploidy results were compared with the genetic results.</p><p><b>RESULTS</b>In CHM, p57(KIP2) negative rates were 95.2% (20/21), whereas all the 11 cases of non-CHM (7 cases PHM and 4 cases hydropic abortions) were positive (11/11). In 11 p57(KIP2) -positive cases, 7 cases with triploidy and 4 cases with diploidy by flow cytometry were proven to be PHM and hydropic abortions by genetic analysis, respectively. Overall, 96.9% (31/32) cases of hydropic placentas were correctly diagnosed by combined use of p57(KIP2) immunohistochemistry and flow cytometry.</p><p><b>CONCLUSIONS</b>p57(KIP2) immunohistochemical negativity is a reliable index for the diagnosis of CHM. Combined flow cytometry DNA ploidy and p57(KIP2) immunohistochemistry are useful in the pathological differentiation of CHM, PHM and hydropic abortions.</p>


Subject(s)
Adult , Female , Humans , Middle Aged , Pregnancy , Young Adult , Abortion, Spontaneous , Diagnosis , Genetics , Metabolism , Cyclin-Dependent Kinase Inhibitor p57 , Metabolism , DNA, Neoplasm , Diagnosis, Differential , Diploidy , Flow Cytometry , Hydatidiform Mole , Diagnosis , Genetics , Metabolism , Immunohistochemistry , Triploidy , Uterine Neoplasms , Diagnosis , Genetics , Metabolism
11.
Article in English | WPRIM | ID: wpr-321110

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expression of imprinted genes related to Beckwith-Wiedemann syndrome (BWS) in human oocytes and preimplantation embryos for understanding the relationship between assisted reproductive technology (ART) and BWS.</p><p><b>METHODS</b>Using nested reverse transcription-PCR to analyze the expression of P57KIP2, LIT1, TSSC3 in human oocytes and preimplantation embryos.</p><p><b>RESULTS</b>Transcripts of P57KIP2 were detected in human oocytes and at all stages of preimplantation embryos. LIT1 was expressed only in stages of 8-cell and blastocyst. Transcripts of TSSC3 could not be detected in human oocytes and preimplantation embryos.</p><p><b>CONCLUSION</b>Transcripts of P57KIP2 and LIT1, imprinted genes related to BWS, were detected in human preimplantation development; ART might affect the epigenetics of imprinted genes in early embryogenesis.</p>


Subject(s)
Female , Humans , Pregnancy , Beckwith-Wiedemann Syndrome , Genetics , Blastocyst , Metabolism , Cyclin-Dependent Kinase Inhibitor p57 , Genetics , Gene Expression Profiling , Genomic Imprinting , Genetics , Nuclear Proteins , Genetics , Oocytes , Metabolism , Potassium Channels, Voltage-Gated , Genetics , Reverse Transcriptase Polymerase Chain Reaction
12.
Article in Korean | WPRIM | ID: wpr-77592

ABSTRACT

BACKGROUND/AIMS: The cyclin-dependent kinase inhibitors (CDKI) including p21, p27, and p57 of the kinase inhibitor protein (KIP) family are negative regulators of cell cycle progression and potentially act as tumor suppressor. Tumor behavior and growth are influenced by the extent of tumor cell proliferation. The aim of this study was to evaluate the expression of KIP family CDKI in gastric cancer tissue, and to examine the relationship between these expression and various clinicopathological parameters including tumor cell proliferation. METHODS: We conducted an immunohistochemical analysis of p21, p27, and p57 expression in 109 gastric cancer tissues. Tumor cell proliferation was assessed by immunohistochemistry with antibody against Ki-67. RESULTS: Negative expression of p21, p27, and p57 was demonstrated in 45.9%, 65.1%, and 57.8% of cancer tissues, respectively. Negative expression of p21 correlated with larger tumor size, poor differentiation, depth of invasion, lymph node metastasis and advanced TNM stage (p=0.048, 0.041, 0.001, 0.005, and 0.001 respectively). Negative expression of p21 correlated with poor survival (p=0.037). Tumors with negative p21 expression had higher Ki-67 expression than those with positive p21 expression (p=0.024). No significant correlation could be observed between status of p27 and p57 expression and various clinicopathological parameters including survival and tumor cell proliferation. CONCLUSIONS: These results suggest that negative expression of p21 may play an important role in carcinogenesis by stimulating tumor cell proliferation, and may help in predicting the prognosis of gastric cancer.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cell Division , Cyclin-Dependent Kinase Inhibitor Proteins/metabolism , Cyclin-Dependent Kinase Inhibitor p21/metabolism , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Cyclin-Dependent Kinase Inhibitor p57/metabolism , English Abstract , Stomach Neoplasms/metabolism , Survival Rate
13.
Chin. med. j ; Chin. med. j;(24): 944-946, 2003.
Article in English | WPRIM | ID: wpr-294197

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of p57(kip2) and cyclinE proteins on the genesis and progression of human pancreatic cancer.</p><p><b>METHODS</b>The expression of p57(kip2) and cyclinE proteins in tumor tissues and adjacent tissues of pancreatic cancer in 32 patients was detected by SP immunohistochemical technique.</p><p><b>RESULTS</b>The p57(kip2) protein positive-expression rate in tumor tissues of pancreatic cancer was 46.9%, which was lower than that in adjacent pancreatic tissue (P < 0.05). The p57(kip2) protein positive-expression correlated significantly with tumor cell differentiation (P < 0.05) and did not correlate significantly with lymph node metastasis (P > 0.05). The cyclinE positive-expression rate in tumor tissues was 68.8%, which was higher than that in adjacent pancreatic tissues (P < 0.05). The cyclinE positive-expression also correlated significantly with tumor cell differentiation and lymph node metastasis (P < 0.05). The cyclinE protein positive-expression rate in the tumor tissues of the p57(kip2) protein positive-expression group was lower than that in the p57(kip2) protein negative-expression group, and there were no significant correlation between the two groups (r = -0.112, P > 0.05).</p><p><b>CONCLUSION</b>Decreased expression of the p57(kip2) protein and/or over-expression of the cyclinE protein may play an important role in the genesis and progression of human pancreatic cancer.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cyclin E , Cyclin-Dependent Kinase Inhibitor p57 , Immunohistochemistry , Nuclear Proteins , Pancreatic Neoplasms , Chemistry , Pathology
14.
Article in Korean | WPRIM | ID: wpr-7484

ABSTRACT

OBJECTIVE: In this study, we investigated the cell growth inhibition, regulation of cell cycle and induction of apoptosis through recombinant p53 adenoviral vector delivery into cervical cancer cell line SiHa, to explore the possibility of p53 gene therapy. METHODS: We infected SiHa with AdCMVp53 at 50 MOI. After 48 hours, the regulation of cell cycle and apoptosis were investigated with FACS. The gene expression profiling associated with cell cycle was also investigated with cell cycle DNA membrane chip. RESULTS: SiHa cells were arrested in the G1 phase by AdCMVp53 and showed cell growth inhibition via apoptosis. The gene expression profiles involved in cell cycle including cyclin-dependent kinase inhibitor 1C (p57, Kip2), RAD9 (S.pombe) homolog, and MAD2 (mitoticarrest deficient, yeast, homolog)-like 2 were up-regulated by more than three-fold, as compared to control group. In contrast, 6 genes such as retinoblastoma-like 2 (p130), and cyclin H were down-regulated by more than three-fold. Several genes known as being differentially up- or down-regulated compared to control were confirmed by RT-PCR and Western blotting assays. CONCLUSION: The adenoviral p53 gene delivery into cervical cancer cell line, suggesting the possibility of p53 gene therapy in cervical neoplasia make the cell growth inhibition and changes of cell cycle-associated gene expression.


Subject(s)
Apoptosis , Blotting, Western , Cell Cycle Proteins , Cell Cycle , Cell Line , Cyclin H , Cyclin-Dependent Kinase Inhibitor p57 , DNA , G1 Phase , Gene Expression , Gene Expression Profiling , Genes, p53 , Membranes , Transcriptome , Uterine Cervical Neoplasms , Yeasts
15.
Chin. med. j ; Chin. med. j;(24): 20-23, 2003.
Article in English | WPRIM | ID: wpr-356877

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the expressions of cyclin E, cyclin dependent kinase 2 (CDK-2) and cyclin-dependent kinase inhibitor p57(KIP2) in human gastric cancer, and to evaluate the relationships between protein levels and clinicopathological parameters.</p><p><b>METHODS</b>Western blot was used to measure the expressions of cyclin E, CDK-2 and p57(KIP2) proteins in the surgically resected gastric carcinoma, adjacent normal mucosa and metastatic lymph nodes from 36 patients.</p><p><b>RESULTS</b>Cyclin E and CDK-2 protein levels were higher in gastric cancer tissues in comparison with normal tissues (P < 0.05). Overexpression of cyclin E was correlated with lymph node involvement, poor histological grade and serosa invasion (P < 0.05). Overexpression of CDK-2 was correlated with lymph nodes involvement (P < 0.05). No statistically significant difference between cyclin E and CDK-2 expression was found when samples were stratified according to tumor size (P > 0.05). Expression of cyclin E and CDK-2 showed a positive linear correlation (r = 0.451, P = 0.01). Protein levels of p57(KIP2) were lower in gastric cancer tissues than in the normal mucosa (P < 0.05). Decreased expression of p57(KIP2) was correlated with lymph node involvement (P < 0.05). No statistically significant difference in p57(KIP2) expression was found when sample were stratified according to tumor size, histological grade or serosa invasion (P > 0.05). In metastatic lymph nodes, expression of cyclin E was increased and the expression of p57(KIP2) decreased.</p><p><b>CONCLUSION</b>Overexpressions of cyclin E, CDK-2 and downregulated expression of p57(KIP2) may play important roles in tumorigenesis and metastatic potential of gastric cancer.</p>


Subject(s)
Humans , Blotting, Western , CDC2-CDC28 Kinases , Cyclin E , Physiology , Cyclin-Dependent Kinase 2 , Cyclin-Dependent Kinase Inhibitor p57 , Cyclin-Dependent Kinases , Physiology , Lymphatic Metastasis , Nuclear Proteins , Physiology , Protein Serine-Threonine Kinases , Physiology , Stomach Neoplasms , Chemistry , Pathology
16.
Article in English | WPRIM | ID: wpr-329165

ABSTRACT

In situ hybridization was applied to locate and detect the expression of p57KIP2 in hydatidiform mole (5 cases of partial hydatidiform mole and 18 cases of complete hydatidiform mole) and normal villi (23 cases). The positive signals of p57KIP2 expression were analyzed by HPIAS-1000 Image-Analysis System. p57KIP2 was highly expressed in normal villi but showed distinct low expression in hydatidiform mole (P < 0.01). Furthermore, the locus of low expression of p57KIP2 accorded with the place where lesion of trophoblast occurred. Detection of p57KIP2 made it possible to study the genetics of hydatidiform mole at the transcriptional level. Low expression of p57KIP2 could be a molecular marker in hydatidiform mole and a target for therapy.


Subject(s)
Female , Humans , Pregnancy , Cyclin-Dependent Kinase Inhibitor p57 , Cyclin-Dependent Kinases , Enzyme Inhibitors , Metabolism , Genomic Imprinting , Genetics , Hydatidiform Mole , Genetics , Metabolism , Nuclear Proteins , Genetics , Placenta , Metabolism , RNA, Messenger , Genetics , Uterine Neoplasms , Genetics , Metabolism
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