Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- ...Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- imens was examined by using polymerase chain reaction-single strand conformation polymorphism (PCR- SSCP) method. The PCR products were sequenced directly. Results: In p21WAF1 /CIP1 gene exon3 of 36 cases of human osteosarcoma, the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 17 cases with the incidence being 44.4%. In 10 normal blood samples, DNA sequence analysis showed the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 8 cases with the incidence being 80%. Conclusion: The novel location of p21WAF1 /CIP1gene polymorphism of osteosarcoma, but not mutation was de?ned, and this location might provide the meaningful reference for the further research of p21WAF1/CIP1 gene.p2lWAF1/CIP1基因DNA序列分析及其与骨肉瘤表型的关系展开更多
AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal ti...AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal tissues from a high risk population for esophageal cancer in northern China were used in this study. All the specimens were fixed with 85% alcohol and further processed with routine histology. The avidin biotin peroxidase complex (ABC) method was used for the detection of p53, Waf1p21 and PCNA. RESULTS The strong nuclear staining for p53, Waf1p21 and PCNA was observed in the normal esophageal epithelium and the epithelia with different severities of lesions. As the lesions progressed to dysplasia (DYS) and to esophageal squamons cell carcinoma (SCC), the percentage of Waf1p21 immunoreactivity decreased. The number of Waf1p21 immunostaining positive cells increased slightly from normal to basal cell hyperplasia (BCH), but there was no further increase in DYS and in SCC. The total number of positive cells for Waf1p21 stain appeared to be lower than that of p53 in normal and BCH esophageal epithelia and much lower in DYS and SCC. The Waf1p21 positive immunostaining cells were located at the third and forth cell layers in half of the samples examined, which was 2～4 cell layers higher than that of PCNA and p53 in the same histological categories of normal, BCH and DYS. CLNCLUSION The low levels of Waf1p21 at the stage of DYS may be related to a functional loss of p53. Other mechanisms may also be responsible to the lack of Waf1p21 expression in DYS and SCC.展开更多
Summary: P21 WAF1/Cip1 , an inhibitor of cyclin dependent kinases, is a critical downstream effector in the P53 specfic pathway of growth control. Increased expression of P21 WAF1/Cip1 has been found to ref...Summary: P21 WAF1/Cip1 , an inhibitor of cyclin dependent kinases, is a critical downstream effector in the P53 specfic pathway of growth control. Increased expression of P21 WAF1/Cip1 has been found to reflect the status of the P53 tumor suppressor pathway. We investigated the expression of P21 WAF1/Cip1 in a relatively small, but well characterized group consisting of 28 hepatocellular carcinomas. The samples were previously studied for P53 gene mutation. P21 WAF1/Cip1 expression were identified by in situ hybridization and immunohistochemistry. Positive ISH for P21 WAF1/Cip1 transcripts was found in 18 of 28 cases (64.3 %). All positive cases by ISH showed detectable P21 WAF1/Cip1 protein reactivity by IHC. No relationship was found between P21 WAF1/Cip1 staining and P53 mutational status. No associations were seen with tumor metastasis, size and tumor grade, except for tumor differentiation status which showed higher frequency of P21 WAF1/Cip1 expression in moderate well differentiated HCCs than poorly differentiated tumors ( P <0.05). It is concluded that expression of P21 WAF1/Cip1 is common in HCCs, but does not correlate with P53 mutational status or pathological parameters investigated except for tumor differentiation. Also, there may be other factors beside P53 that regulate P21 WAF1/Cip1 gene expression in HCCs.展开更多
INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced b...INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced by hepatitis B virus (HBV) and/oraflatoxin B1 (AFB1).展开更多
AIM:To investigate the relationship between expression of p21^(WAF1) and p53 gene,and to evaluate the deletion and polymorphism of p21^(WAF1) gene in gastric carcinoma (GC). METHODS:Expression of p21 and p53 proteins,...AIM:To investigate the relationship between expression of p21^(WAF1) and p53 gene,and to evaluate the deletion and polymorphism of p21^(WAF1) gene in gastric carcinoma (GC). METHODS:Expression of p21 and p53 proteins,and deletion and polymorphism of p21 gene in GC were examined by streptavidin-peroxidase conjugated method (SP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) respectively. RESULTS:The expression of p21 and p53 was found in 100% (20/20) and 0% (0/20) of normal gastric mucosae (NGM),92.5% (37/40) and 15.0% (6/40) of dysplasia (DP) and 39.8% (43/108) and 56.5% (61/108) of GC,respectively. The positive rate of p21 in GC was lower than that in NGM and DP (P<0.05),while the positive rate of p53 in GC was higher than that in NGM end DP (P<0.05).p21 and p53 were significantly expressed in 63.3% (19/30) and 36.7% (11/30),35.0% (14/40) and 77.5% (31/40),26.7% (4/15) and 80.0% (12/15),30.8% (4/13) and 30.8% (4/13),and 20.0% (2/10) and 30.0%,(3/10) of well-differentiated, poorly-differentiated,undifferentiated carcinomas,mucoid carcinomas and signet ring cell carcinomas.The expression of p21 in well-differentiated carcinomas was significantly higher than that in poorly-differentiated,un-differentiated, mucoid carcinomas and signet ring cell carcinomas (P<0.05). Contrarily,The expression of p53 was increased from well- differentiated to poorly-differentiated and un-differentiated carcinomas (P<0.05).The expression of p21 and p53 in paired primary and metastatic GC (35.3% and 70.6%) was different from non-metastatic GC (62.5% and 42.5%) markedly (P<0.05).The expression of p21 in invasive superficial muscle (60.0%) was higher than that in invasive deep muscle or total layer (35.2%) (P<0.05) and was higher in TNM stages Ⅰ (60.0%) and Ⅱ (56.2%) than in stages Ⅲ (27.9%) and Ⅳ (22.2%) (P<0.05),whereas the expression of p53 did not correlate to invasion depth or TNM staging (P>0.05).The exoression patterns of p53+/p21-,and of p53-/p21+ were found in 5.0% and 82.5% of DP.There was a significant correlation between expression of p21 and p53 (P<0.05).But there was no significant correlation between expression of both in GC (P>0.05).There was no deletion in exon 2 of p21 gene in 30 cases of GC and 45 cases of non-GC,but polymorphism of p21 gene at exon 2 was found in 26.7% (8/30) of GC and 8.9% (4/45) of non- GC,a significant difference was found between GC and non-GC (P<0.05).There was no significant relation between p21 expression of polymorphism (37.5%,3/8) and non-polymorphism (45.5%,10/22) in GC (P>0.05). CONCLUSION:The loss of p21 protein and abnormal expression of p53 are related to carcinogenesis,differentiation and metastasis of GC.The expression of p21 is related to invasion and clinical staging in GC intimately.The expression of p21 protein depends on p53 protein largely in NGM and DP,but not in GC.No deletion of p21 gene in exon 2 can be found in GC.The polymorphism of p21 gene might be involved in gastric carcinogenesis.There is no significant association between polymorphism of p21 gene and expression of p21 protein.展开更多
Background Mutation and expression change of p21 WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21 WAF1/CIP1 gene in human osteosarcoma,p21 WAF1/...Background Mutation and expression change of p21 WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21 WAF1/CIP1 gene in human osteosarcoma,p21 WAF1/CIP1 gene DNA sequence change and their relationships with the phenotype and clinical prognosis.Methods p21 WAF1/CIP1 gene in 10 normal people and the tumours of 45 osteosarcoma patients were examined using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) with silver staining. The PCR product with an abnormal strand was sequenced directly. The p21 WAF1/CIP1 gene mRNA and P21 protein of 45 cases of osteosarcoma were investigated by using in situ hybridization and immunohistochemistry,respectively. Results The occurrence of P21 protein in osteosarcoma was 17.78% (8/45),and p21 WAF1/CIP1 mRNA expression in osteosarcoma was 42.22% (19/45). The p21 WAF1/CIP1 gene DNA sequencing of amplified production showed that in p21 WAF1/CIP1 gene exon 3 of 36 cases of human osteosarcoma,there were 17 cases (47.22%) with C→T at position 609; 10 normal blood samples’ DNA sequence analysis yielded 8 cases (80.00%) with C→T at the same position. Conclusions Along with the increase of malignancy,the expression of p21 WAF1/CIP1 mRNA and P21 protein in osteosarcoma tends to decrease. It is uncommon for the p21 WAF1/CIP1 gene mutation to occur in human osteosarcoma. As a result, the possible existence of tumour subtypes of p21 WAF1/CIP1 gene mutation should be investigated. Our research leads to the location of p21 WAF1/CIP1 gene polymorphism of Chinese osteosarcoma patients,which can provide a basis for further research.展开更多
文摘Objective: To investigate the p21WAF1 /CIP1gene DNA sequence change and their relationship with the phenotype of human osteosarcoma. Methods: p21WAF1 /CIP1gene DNA of 36 osteosarcoma spec- imens was examined by using polymerase chain reaction-single strand conformation polymorphism (PCR- SSCP) method. The PCR products were sequenced directly. Results: In p21WAF1 /CIP1 gene exon3 of 36 cases of human osteosarcoma, the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 17 cases with the incidence being 44.4%. In 10 normal blood samples, DNA sequence analysis showed the change of C→T in the p21WAF1 /CIP1gene CDNA sequence of position 609th occurred in 8 cases with the incidence being 80%. Conclusion: The novel location of p21WAF1 /CIP1gene polymorphism of osteosarcoma, but not mutation was de?ned, and this location might provide the meaningful reference for the further research of p21WAF1/CIP1 gene.p2lWAF1/CIP1基因DNA序列分析及其与骨肉瘤表型的关系
文摘AIM To study the correlationship between the changes of p53, Waf1p21 and the cell proliferation determined by PCNA at different stages of human esophageal carcinogenesis. METHODS Biopsied and resected esophageal tissues from a high risk population for esophageal cancer in northern China were used in this study. All the specimens were fixed with 85% alcohol and further processed with routine histology. The avidin biotin peroxidase complex (ABC) method was used for the detection of p53, Waf1p21 and PCNA. RESULTS The strong nuclear staining for p53, Waf1p21 and PCNA was observed in the normal esophageal epithelium and the epithelia with different severities of lesions. As the lesions progressed to dysplasia (DYS) and to esophageal squamons cell carcinoma (SCC), the percentage of Waf1p21 immunoreactivity decreased. The number of Waf1p21 immunostaining positive cells increased slightly from normal to basal cell hyperplasia (BCH), but there was no further increase in DYS and in SCC. The total number of positive cells for Waf1p21 stain appeared to be lower than that of p53 in normal and BCH esophageal epithelia and much lower in DYS and SCC. The Waf1p21 positive immunostaining cells were located at the third and forth cell layers in half of the samples examined, which was 2～4 cell layers higher than that of PCNA and p53 in the same histological categories of normal, BCH and DYS. CLNCLUSION The low levels of Waf1p21 at the stage of DYS may be related to a functional loss of p53. Other mechanisms may also be responsible to the lack of Waf1p21 expression in DYS and SCC.
文摘Summary: P21 WAF1/Cip1 , an inhibitor of cyclin dependent kinases, is a critical downstream effector in the P53 specfic pathway of growth control. Increased expression of P21 WAF1/Cip1 has been found to reflect the status of the P53 tumor suppressor pathway. We investigated the expression of P21 WAF1/Cip1 in a relatively small, but well characterized group consisting of 28 hepatocellular carcinomas. The samples were previously studied for P53 gene mutation. P21 WAF1/Cip1 expression were identified by in situ hybridization and immunohistochemistry. Positive ISH for P21 WAF1/Cip1 transcripts was found in 18 of 28 cases (64.3 %). All positive cases by ISH showed detectable P21 WAF1/Cip1 protein reactivity by IHC. No relationship was found between P21 WAF1/Cip1 staining and P53 mutational status. No associations were seen with tumor metastasis, size and tumor grade, except for tumor differentiation status which showed higher frequency of P21 WAF1/Cip1 expression in moderate well differentiated HCCs than poorly differentiated tumors ( P <0.05). It is concluded that expression of P21 WAF1/Cip1 is common in HCCs, but does not correlate with P53 mutational status or pathological parameters investigated except for tumor differentiation. Also, there may be other factors beside P53 that regulate P21 WAF1/Cip1 gene expression in HCCs.
基金the National Natural Science Foundation of China,No.39260033.
文摘INTRODUCTIONIn order to study the relationship between oncogeneexpression and HCC generation,we observed theprecancerous hepatic GGT loci,IGF-Ⅱ,p53 andp21 expression during hepatocarcinogenesis of treeshrew induced by hepatitis B virus (HBV) and/oraflatoxin B1 (AFB1).
基金Supported by the Key Programs during the 8~(th)5-Year Plan Period,the Bureau of Health,Hunan Province, China, No. 9301
文摘AIM:To investigate the relationship between expression of p21^(WAF1) and p53 gene,and to evaluate the deletion and polymorphism of p21^(WAF1) gene in gastric carcinoma (GC). METHODS:Expression of p21 and p53 proteins,and deletion and polymorphism of p21 gene in GC were examined by streptavidin-peroxidase conjugated method (SP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) respectively. RESULTS:The expression of p21 and p53 was found in 100% (20/20) and 0% (0/20) of normal gastric mucosae (NGM),92.5% (37/40) and 15.0% (6/40) of dysplasia (DP) and 39.8% (43/108) and 56.5% (61/108) of GC,respectively. The positive rate of p21 in GC was lower than that in NGM and DP (P<0.05),while the positive rate of p53 in GC was higher than that in NGM end DP (P<0.05).p21 and p53 were significantly expressed in 63.3% (19/30) and 36.7% (11/30),35.0% (14/40) and 77.5% (31/40),26.7% (4/15) and 80.0% (12/15),30.8% (4/13) and 30.8% (4/13),and 20.0% (2/10) and 30.0%,(3/10) of well-differentiated, poorly-differentiated,undifferentiated carcinomas,mucoid carcinomas and signet ring cell carcinomas.The expression of p21 in well-differentiated carcinomas was significantly higher than that in poorly-differentiated,un-differentiated, mucoid carcinomas and signet ring cell carcinomas (P<0.05). Contrarily,The expression of p53 was increased from well- differentiated to poorly-differentiated and un-differentiated carcinomas (P<0.05).The expression of p21 and p53 in paired primary and metastatic GC (35.3% and 70.6%) was different from non-metastatic GC (62.5% and 42.5%) markedly (P<0.05).The expression of p21 in invasive superficial muscle (60.0%) was higher than that in invasive deep muscle or total layer (35.2%) (P<0.05) and was higher in TNM stages Ⅰ (60.0%) and Ⅱ (56.2%) than in stages Ⅲ (27.9%) and Ⅳ (22.2%) (P<0.05),whereas the expression of p53 did not correlate to invasion depth or TNM staging (P>0.05).The exoression patterns of p53+/p21-,and of p53-/p21+ were found in 5.0% and 82.5% of DP.There was a significant correlation between expression of p21 and p53 (P<0.05).But there was no significant correlation between expression of both in GC (P>0.05).There was no deletion in exon 2 of p21 gene in 30 cases of GC and 45 cases of non-GC,but polymorphism of p21 gene at exon 2 was found in 26.7% (8/30) of GC and 8.9% (4/45) of non- GC,a significant difference was found between GC and non-GC (P<0.05).There was no significant relation between p21 expression of polymorphism (37.5%,3/8) and non-polymorphism (45.5%,10/22) in GC (P>0.05). CONCLUSION:The loss of p21 protein and abnormal expression of p53 are related to carcinogenesis,differentiation and metastasis of GC.The expression of p21 is related to invasion and clinical staging in GC intimately.The expression of p21 protein depends on p53 protein largely in NGM and DP,but not in GC.No deletion of p21 gene in exon 2 can be found in GC.The polymorphism of p21 gene might be involved in gastric carcinogenesis.There is no significant association between polymorphism of p21 gene and expression of p21 protein.
基金ThisstudywassupportedbytheChineseNationalInstituteofHealthGrant (No 96 13 7)
文摘Background Mutation and expression change of p21 WAF1/CIP1 may play a role in the growth of osteosarcoma. This study was to investigate the expression of the p21 WAF1/CIP1 gene in human osteosarcoma,p21 WAF1/CIP1 gene DNA sequence change and their relationships with the phenotype and clinical prognosis.Methods p21 WAF1/CIP1 gene in 10 normal people and the tumours of 45 osteosarcoma patients were examined using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) with silver staining. The PCR product with an abnormal strand was sequenced directly. The p21 WAF1/CIP1 gene mRNA and P21 protein of 45 cases of osteosarcoma were investigated by using in situ hybridization and immunohistochemistry,respectively. Results The occurrence of P21 protein in osteosarcoma was 17.78% (8/45),and p21 WAF1/CIP1 mRNA expression in osteosarcoma was 42.22% (19/45). The p21 WAF1/CIP1 gene DNA sequencing of amplified production showed that in p21 WAF1/CIP1 gene exon 3 of 36 cases of human osteosarcoma,there were 17 cases (47.22%) with C→T at position 609; 10 normal blood samples’ DNA sequence analysis yielded 8 cases (80.00%) with C→T at the same position. Conclusions Along with the increase of malignancy,the expression of p21 WAF1/CIP1 mRNA and P21 protein in osteosarcoma tends to decrease. It is uncommon for the p21 WAF1/CIP1 gene mutation to occur in human osteosarcoma. As a result, the possible existence of tumour subtypes of p21 WAF1/CIP1 gene mutation should be investigated. Our research leads to the location of p21 WAF1/CIP1 gene polymorphism of Chinese osteosarcoma patients,which can provide a basis for further research.
