Objective To observe the expression of p53, bcl-2 genes, vascular endothelial cell growth factor（VEGF）, basic fibroblast growth factor（bFGF）, insulin-like growth factor-I （IGF-I）, and the receptors of these factors of retinal vascular endothelial cells (VECs) of 1- to 20-week diabetic rats, and the relationship between the expressions and cell cycle arrest.Methods Retinal sections of diabetic rats induced by alloxan were immunohistochemically stained and observed by light microscopy (LM) and electron microscopy (EM). Dot blotting and Western blotting were used to determine the expression of mRNA, proteins of p53 and bcl-2. Results Under LM, immunohistochemical positive expression of p53 and bcl-2 were found on the vessels of ganglion cell layer and inner nuclear layer of retinae of 8- to 20-week diabetic rats; under EM, these substances were observed depositing in VECs. The retinal VECs also expressed VEGF, bFGF, IGF-I and their receptors. There was no positive expression of other cell types in these retinae, all cell types of retinae in control group, or all cells of retinae of diabetic rats with the course of disease of 1 to 6 weeks. The result of dot blotting revealed that retinal tissue of 20-week diabetic rat expressed p53 and bcl-2 mRNA, and the result of Western blotting revealed that they also expressed p53 and bcl-2 proteins. But retinal tissues of control group did not. Positive expression of bax was not found in the retinae in control group or 1- to 20-week diabetic rats. Conclusion p53, bcl-2 may introduce cell cycle arrest of VECs of retinae in 8- to 20-week diabetic rats. High glucose might stimulate the expression of VEGF, bFGF, IGF-I and their receptors, and the growth factors may keep VECs surviving by self-secretion. (Chin J Ocul Fundus Dis，2003，19：29-33)
【Abstract】Objective To investigate the association between p53, c-erb B-2 oncogene protein and angiogenesis in breast carcinoma and breast with atypical hyperplasia. Methods Immunohistochemical reaction of p53, c-erb B-2 oncogene protein was evaluated in 103 benign and malignant lesion breast paraffin-embedded specimens. The microvessel endothelial area (MEA) was quantitated by computer image analysis system (CIAS), which was immunohistochemically stained by FⅧ-RA. The relationships between p53, c-erb B-2 and MEA were analyzed. Results The MEA of p53 oncogene protein positive expression in mild atypical hyperplasia breast lesion was significantly higher than that in p53 negative (t=2.302 4,P＜0.05). The MEA of p53 oncogene protein positive expression in severe atypical hyperplasia was higher than that in p53 negative (t=2.179 4,P＜0.05). Moreover, no significant association between c-erb B-2 oncogene protein and MEA was observed. Conclusion p53 oncogene mutant,protein expression is significantly related to angiogenesis.
Objective To investigate the relationship between p53 codon 72 polymorphism and susceptibility to keloid. Methods The p53 genotypes were detected by polymerase chain reactionreverse dot blot(PCRRDB) and DNA direct sequencing among 15 healthy controls and 15 patients with keloid. Results The frequency of the Proallele(P=0.035) and Pro/Pro genotype(P=0.030) in patients was significantly higher than that in the controlls. There was no significant difference in the frequency of Pro/Arg and Arg/Arg genotypes between patients and controls. Conclusion The p53 gene codon 72 polymorphism may play a role in susceptibility to keloid.
Objective To explore the effect of exogenous estrogen receptor β1 (ERβ1) gene on the expression of p53 as well as the changes of apoptosis in MDA-MB-231 cell line and to investigate the biological role of ERβ1 in breast cancer. Methods Recombinant eukaryotic expressing vector containing ERβ1 cDNA was transfected into human breast cancer cell MDA-MB-231 by using cationic liposome LipofectamineTM 2000. The expression levels of p53 and ERβ1 in mRNA and protein were evaluated by real-time PCR and Western blot, respectively. Cell growth curve was used to detect the changes of cell proliferation ability. Cell apoptosis was detected by flow cytometry. Results After transfected with vector containing ERβ1 cDNA, proliferation ability of MDA-MB-231 cell decreased and the expression levels of both ERβ1 and p53 in both mRNA and protein increased (Plt;0.01). Rate of cell apoptosis increased in ERβ1 upregulation groups （Plt;0.01）. Conclusion ERβ1 can induce apoptosis and inhibit the growth of MDA-MB-231 cells by upregulating p53 expression.
Objective To clarify the correlation of p53 codon 72 polymorphism in peripheral blood with keloid susceptibility in Chinese population. Methods All the literatures of case-control research on the correlation between p53 codon 72 polymorphism in peripheral blood and keloid in Chinese population were searched in PubMed, EBSCO, CNKI, CBM, and WanFang Data from their establishment to August 2010. Meta-analyses were performed to detect whether there were differences between the keloid group and the control group about the distribution of genotypes of p53 codon 72 in peripheral blood, such as, Pro/Pro vs. Arg/Arg, Pro/Pro vs. Pro/Arg, and alleles Pro vs. Arg. Results Five studies involving 328 keloid patients and 420 patients in the control group were included. The results of meta-analyses showed that the population having the genotype Pro/Pro presented no increased keloid risk compared to that with the genotypes Arg/Arg (OR=2.17, 95%CI 0.86 to 5.47) or Pro/Arg (OR=1.90, 95%CI 0.92 to 3.93), while the allele Pro showed significant association with increased keloid risk compared to the allele Arg (OR=1.86, 95%CI 1.03 to 3.35). Conclusion The allele Pro of p53 codon 72 in peripheral blood of Chinese population is significantly associated with increased keloid risk.
ObjectiveTo explore the relationship between p53 mutation in 5-8 exons and type of epithelial ovarian cancer (EOC) pathogenesis of Han nationality women. MethodsFrom August 2011 to December 2012, 45 patients with primary EOC (Han nationality women from Sichuan Province) diagnosed surgically and pathologically were selected. Using direct DNA sequencing, we analyzed the mutations of p53 in 5-8 exons of all cases, and the EOC patients were divided into two types according dualism and the pathogenesis results. The p53 mutation of the different types in EOC patients were analyzed. ResultsThe frequency and efficiency of p53 mutation in type-ⅡEOC patients were significantly higher than that in typeⅠ(P < 0.01). And the codon 175 might be a mutational hotspot of type-ⅡEOC. The malignant degree and oviduct involved frequency of type-ⅡEOC were obviously higher than that of type-I EOC; p53 mutation frequency in high malignant patients increased significantly. Conclusionsp53 mutation plays an important role in the development of type-ⅡEOC. The codon 175 might be a mutational hotspot of type-ⅡEOC.
ObjectiveTo observe the expression and mechanism of hypoxia-inducible factor-1α (HIF-1α) and p53 protein at the altitude of 5000 meter plateau hypoxia environment in rats, as well as the effect of Astragalus injection. MethodsSixty Sprague Dawley rats were randomly divided into the Astragalus injection intervention group and normal saline control group, 30 rats in each group. Astragalus injection group rats were intraperitoneal injected of Astragalus injection (15 ml/kg) before 30 minutes into the plateau environment simulation cabin, normal saline group rats were intraperitoneal injected with the same volume of saline. 30 minutes after injection, rats in each group were reared in the plateau experiment cabin which simulated altitude of 5000 m (oxygen partial pressure 11.3 kPa) for 2, 6, 8, 12, 24 hours, each time period of 6 rats. When get out, the rats were executed immediately and eyes were harvested. Retinal sections were studied by hematoxylin eosin stain, and immunohistochemical method for HIF-1α and p53 expression. ResultsFor control rats, after 2 hours in the cabin, there was edema in retinal layers. HIF-1α and p53 were expressed mainly in the cytoplasm of retinal layers. When the periods in cabin extended, there was atrophy of retinal nerve fiber layer, swelling and degeneration of ganglion cells. The expression of HIF-1α and p53 was increased. Compared with the control group, the intervention group rat had similar but less severe retinal changes, and the expression of HIF-1α and p53 was significantly decreased (P<0.05). ConclusionAstragalus injection can reduce pathological retinal damage in rats at high altitude environment, and its mechanism may be associated with reduced HIF-1α, p53 expression.
ObjectiveTo study the relationship between mutation of the p53 gene, p21 gene and bacteria Lform in gallbladder carcinoma. MethodsForty cases of gallbladder carcinoma and 40 cases of chronic cholecystitis were studied by using Gram staining and immunohistochemistry (SP method) to detect the positive rate of Lforms antigen, p21 and p53 protein overpression. And the relationship between the expression of p21 and p53 in Lform infection positive group and that in Lform infection negative group was discussed. ResultsThere was no statistical difference between the Lform positive rate in patients with gallbladder carcinoma with Gram staining and immunohistochemistry (Pgt;0.05). The positive expression rate of p21 and p53 in gallbladder carcinoma was 62.5%(25/40) and 65.0%(26/40) respectively. The expression values of p21 and p53 in chronic cholecystitis was 2.5%(1/40) and 5.0%(2/40) respectively, which was significantly different from that of gallbladder carcinoma (P＜0.05). The expression of p21 and p53 was significantly higher in Lform infection positive group than in that with Lform infection negative group (P＜0.05). ConclusionBacteria Lform may be one of the direct factor leading to mutation of p53 and p21 during gallbladder oncogenesis.
Objective To study the effect of 5-fluorouracil (5-FU) induced apoptosis of the rectal carcinoma cell line HR8348 in vitro and the relationship between apoptosis induced by 5-FU and the expression of bcl-2，bcl-xl，bax and p53,and to investigate the possible mechanism of apoptosis of rectal carcinoma cell line HR8348 induced by 5-FU.Methods After treatment with 5-FU for 24 h,the apoptotic index was detected by methyl green and pyronine Y staining and by terminal deoxynucleotidyl transferase (TdT)mediated dUTP nick end labeling (TUNEL).The bcl-2,bcl-xl,bax and p53 gene expression of HR8348 cells were examined by immunohistochemical method.Results After treatment with 5-FU,the apoptotic index of experiment group was significantly increased,there was significant difference as compared with the control.Exposed to 5-FU for 12 h,24 h and 36 h,the expression of bcl-2 of HR8348 cell line remained unchanged,but the expression of bcl-xl slightly diminished,while the expression of bax was remarkly increased,the expression of p53 was not detected in both experiment and control groups.Conclusion This results indicate that 5-FU may induce apoptosis of rectal carcinoma cell line HR8348 and the possible mechanism of apoptosis induction is through upregulation of bax expression and the change of bax to bcl-xl ratio.
【Abstract】Objective To investigate the expression of inducible nitric oxide synthase (iNOS) and p53 protein in hepatocellular carcinoma (HCC) and their relationship with angiogenesis. Methods Immunohistochemical method and image analysis technique were used to detect the expression of iNOS and p53 protein in tumor tissue sections of 59 HCC patients. Microvessel density (MVD) was counted by immunohistochemical staining with anti-CD34 antibody.Results ①The expression rates of iNOS and p53 were 81.4%(48/59), 64.4%(38/59) in HCC patients, respectively. The expression intensities of iNOS and p53 were 5 635±1 287, 3 352±873 in HCC patients, respectively. ②MVD was 32.5±2.73 in the tumor tissue of HCC patients. ③The expression of iNOS was correlated with the expression of p53 and MVD in HCC patients (P＜0.05); The expression of p53 was also correlated with the MVD in HCC patients (P＜0.05). Conclusion iNOS and p53 are highly expressed in HCC and may play a key role in angiogenesis of HCC.