Objective To investigate the possible interaction between the ras and p53 genes overexpression in thyroid carcinoma, and whether there is correlation between the ras and p53 overexpression and clinico-pathological criteria. Methods Thyroid lesions from eighty patients were examined for expression of ras and p53 genes by the LSAB immunohistochemistic method. Of these patients, 54 were diagnosed as malignant lesions and 26 benign nodular thyroid disorders. Results The positive immunostain rate for ras and p53 genes was 90.7%, 23.0% and 55.5%, 30.7% in carcinoma and benign lesions respectively with statistically significance between thyroid carcinomas and benign disorders (Ｐ＜0.05). Both ras and p53 overexpressions coexisted in 30 thyroid carcinomas and follow-up showed that 3 of them died and 5 of them had recurrence within 4 years.Conclusion Activation of ras gene and inactivation of p53 gene are cooperatively associated in thyroid tumorigenesis. The concurrent overexpression of ras and p53 could result in a poor prognosis.
Objective To explore the expression of survivin gene in retinoblastoma (RB) and its relationship with the stages and histodifferentiation degree of RB and the expression of p53、bcl-2 proteins. Methods Expression of survivin, p53 and bcl-2 proteins in 38 RB conventional paraffin samples were detected with survivin, p53 and bcl-2 monoclonal antibodies respectively by immunohistochemical assay. The expression of survivin of normal retina in 6 control samples was observed. Results In 38 cases of RB, positive expression of survivin was found in 20 (52.6%); while none of the 6 normal retinal tissue expressed survivin, which had significant difference between the two group (P＜0.05). The positive expression of survivin did not correlate with sex of patient, disease stages and histological type (P＞0.05). In 38 RB cases, positive expression of p53 was in 25 with the rate of 65.8%, and of bcl-2 in 18 with the rate of 47.4%. The positive-expressed rates were much higher in positive-expressed p53 and bcl-2 group than those in the negative-expressed p53 and bcl-2 group(P＜0.05). Conclusion The increase of the expression of survivin implies that it may take part in the occurrence and development of RB; the interaction among survivin, p53 and bcl-2 may participate in the access and the course of RB. (Chin J Ocul Fundus Dis，2004，20：215-217)
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.
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 establish a better immunofluorescence protocol to detect co-localization of p53 and mitochondria which may benefit studies aiming to detect mitochondrial expression of proteins.MethodsHeLa cells were treated with hypoxia and the expression of p53 was detected by immunoblotting. HeLa cells were fixed with methanol, methanol: acetone (1: 1, v/v) mixture, and 4% paraformaldehyde, respectively; the former two groups were not permeable, while the latter was penetrated with 0.1% Triton-X 100 and stained with p53 and mitochondria at the same time. After HeLa cells were fixed with 4% paraformaldehyde, the concentration of Triton-X 100 was reduced to 0.05%, 0.025%, 0.01%, and 0.005%. After the HeLa cells were fixed with 4% paraformaldehyde, the concentration of Triton-X 100 decreased to 0.01% and 0.005% for the first time, then, after staining with p53, the mitochondria were stained with 0.1% Triton-X 100 for the second time.ResultsThe expression of p53 was up-regulated (P<0.01) after hypoxia, which could be used in the following immunofluorescence experiment. The co-localization of p53 and mitochondria was observed in the nucleus and cytoplasm in both the methanol group and the mixed solution group. The co-localization of p53 was the most obvious in the mixed solution group. After using 0.1% Triton-X 100, the p53 signal was mainly in the nucleus, but no co-localization was observed. After fixation with 4% paraformaldehyde, to some extent, the reduced concentration of 0.05% and 0.025% Triton-X 100 weakened the p53 signal in nucleus and enhanced the co-localization signal. However, the signal in nucleus was still stronger than that in cytoplasm. When it was reduced to 0.01% and 0.005%, p53 signal was detected in cytoplasm but not in nucleus, suggesting that the nuclear membrane was not penetrated under this condition, but it also failed to penetrate the mitochondrial membrane, leading to the failure of mitochondrial labeling. The second permeability completely avoided the p53 signal in nucleus, and successfully labeled mitochondria, and the co-localization of p53 and mitochondria was detected.ConclusionsCo-localization of p53 and mitochondria is detectable in cells fixed by methanol or methanol and acetone mixture which brings out better results. Penetrating twice with Triton X-100 of different concentrations following paraformaldehyde fixation help avoid signals in nuclei and falicitate co-localization detection.
Objective To investigate the proportions of CD4+ T cells, CD8+ T cells, and mutant of p53 gene in the microenvironment of breast infiltrating ductal carcinoma, and to explore its’ correlation with prognosis of breast infiltrating ductal carcinoma. Methods Eighty-five cases of breast infiltrating ductal carcinoma were collected who underwent surgery in the 371st Central Hospital of Peoples’ Liberation Army from 2010 to 2012, and then detected the proportion of CD4+ T cells and CD8+ T cells, ratio of CD4+ T cells to CD8+ T cells, and mutant of p53 gene in the cancer tissues with immunohistochemistry. Comparison between the sentinel lymph node metastasis group and non-sentinel lymph node metastasis group, mutant of p53 gene group and non-mutant of p53 gene group on the proportions of CD4+ T cells, CD8+ T cells, and ratio of CD4+ T cells to CD8+ T cells were performed, as well as the relationship between proportion of CD8+ T cells/mutant of p53 gene and prognosis of breast infiltrating ductal carcinoma. Results ① The relationship between proportion of CD4+ T cells/proportion of CD8+ T cells/ratio of CD4+ T cells to CD8+ T cells and situation of sentinel lymph node metastasis: at cluster, compared with the sentinel lymph node metastasis group, the proportion of CD8+ T cells was lower in the non-sentinel lymph node metastasis group (P<0.05), but there was no significant difference on the proportion of CD4+ T cells and ratio of CD4+ T cells to CD8+ T cells (P>0.05); at stroma, compared with the sentinel lymph node metastasis group, the proportions of CD4+ T cells and CD8+ T cells were lower, but the ratio of CD4+ T cells to CD8+ T cells was higher in the non-sentinel lymph node metastasis group (P<0.05). ② The relationship between proportion of CD4+ T cells/proportion of CD8+ T cells/ratio of CD4+ T cells to CD8+ T cells and mutant of p53 gene: both at the cluster and stroma, compared with the mutant of p53 gene group, the proportions of CD4+ T cells and CD8+ T cells were lower, but the ratio of CD4+ T cells to CD8+ T cells was higher in the non-mutant of p53 gene group (P<0.05). ③ The relationship between proportion of CD8+ T cells/mutant of p53 gene and prognosis of breast infiltrating ductal carcinoma: the prognosis was worse in patients with high degree of infiltration of CD8+ T cells and mutant of p53 gene than those patients with low degree of infiltration of CD8+ T cells and non-mutant of p53 gene (P<0.05). Conclusions The proportions of CD4+ T cells and CD8+ T cells, and ratio of CD4+ T cells to CD8+ T cells are associated with the situation of sentinel lymph node metastasis and mutant of p53 gene, and the degree of infiltration of CD8+ T cells and mutant of p53 gene are associated with the prognosis of breast infiltrating ductal carcinoma.
【Abstract】ObjectiveTo study the expressions of P-gp, nm23 and p53 in primary breast cancer tissues and to evaluate the prediction significance in recurrence of the primary breast cancer. MethodsExpressions of P-gp, nm23 and p53 in 57 benign and malignant breast paraffin-embedded specimens, the difference of the rate and intensity of positive reaction of P-gp,nm23 and p53 were checked by immunohistochemical staining.ResultsThe positive rate of nm23 significantly increased in benign breast tissues compared with the tissues of recurrence of breast cancer. The positive intensity of nm23 was significantly decreased in the tissues of recurrence of breast cancer compared with the tissues of primary breast cancer. The positive rate of p53 was significantly increased in malignant breast tissues compared with benign breast tissues. The positive intensity of p53 was significantly increased in the tissues of recurrence of breast cancer compared with the tissues of primary breast cancer. There was no difference in the positive rate of P-gp in the tissue of recurrence and primary of breast cancer compared with benign breast tumor. But the positive intensity of P-gp was significantly decreased in the tissues of recurrence and primary of breast cancer compared with benign breast tumor. ConclusionThe expression of p53 may indicate the proliferating activity of carcinoma cell. The positive intensity of expression of p53 and nm23 is valuable in prediction of the recurrence of primary breast cancer.
ObjectiveTo investigate the relationship between p53 gene family and thyroid cancer.MethodThe related literatures in the database were reviewed and analyzed.ResultsThe p53 gene family included p53, p63, and p73. The p53 played an important role in the development of thyroid cancer, especially in the development of undifferentiated thyroid cancer. The different subtypes of p63 might have different roles in the thyroid cancer, so it couldn’t be generally said that the p63 was an oncogene or an anti-oncogene, and the function of its specific protein needed to be further studied. The biological role of p73 in the thyroid cells might be contradictory, depending on the interaction of many different factors, and the interaction between various p73 subtypes and members of the p53 molecular network was still unclear.ConclusionThere is still some controversy about role of p53 gene family in development of thyroid cancer.
【Abstract】Objective To investigate whether SUMO-1 enhances the apoptosis induced by wild-type p53 plasmid transfection in HepG2 cells. Methods The HepG2 cells were transfected respectively or simultaneouly with the following expressional plasmids as pcDNA3-wtp53（pwtp53，including human wild-type p53 gene），pCMV-HDM1B（pMDM2，including HDM2 gene， homologous gene as murine double minute gene 2），pcDNA3-His6-SUMO-1（pSUMO-1 ，including small ubiquitin-like modifier1 gene）and plasmid pcDNA3.The proteins expressed in cells were detected by means of Western blotting and the apoptosis rates of cells were measured by flow cytometry. Results The protein bands of p53 and MDM2 could be seen in cells transfected with pwtp53 and pMDM2. Meanwhile，the relative larger molecular weight bands were also seen in cells transfected with pSUMO-1 which represented the p53 and MDM2 protein modification by SUMO-1. Merely the trace of p53 protein was detected in cells not transfected with any plasmid or only transfected with empty plasmid and pSUMO-1. In cells transfected with pwtp53 and pwtp53+pSUMO-1，the apoptosis rates were (16.79±1.62)%and (18.15±1.36)%. When transfected with pwtp53+pMDM2，the rate decreased to (5.17±1.23)%. The apoptosis rate would come up again to (14.06±1.84)% after transfected with pwtp53+pMDM2+pSUMO-1 and the difference of rates were significant compared to the cells transfected with pwtp53+pMDM2 (PH＜0.01）. The apoptosis rates in other cells were less than 2% and had no significant difference. Conclusion SUMO-1 could increase the apoptosis induced by wild-type p53 plasmid transfection in HepG2 cells through combining to p53 protein or its post-translational modification and inhibiting p53 degradation by MDM2.