Relationship between therapeutic efficacy of arterial infusion chemotherapy and expression of P-glycoprotein and p53 protein in advanced hepatocellular carcinoma

AIM： To investigate the relationship between the chemotherapeutic drug efficacy and the expression of P-glycoprotein （PGP） and p53 protein in advanced hepatocellular carcinoma （HCC）. METHODS： The study was conducted on 41 patients with advanced HCC who were treated by repeated arterial infusion chemotherapy. Biopsy specimens from the tumor were collected before the start of treatment in all the patients, and the specimens were stored frozen until immunohistochemical staining, which was performed after the start of treatment, to detect PGP and p53 protein expressions. Twenty of the fortyone patients were treated with an anthracycline drug （epirubicin hydrochloride; anthracycline group）, and the remaining 21 were treated with a non-anthracycline drug （mitoxantrone hydrochloride in 11 patients and carboplatin in 10 patients; non-anthracycline group）. The relationship between the chemotherapeutic efficacy and the results of immunostaining were compared between the two groups. RESULTS： Before the start of the treatment, PGPpositive rate was 90.2% （strongly-positive, 36.6%） and p53 protein-positive rate was 34.1% （strongly-positive, 19.5%）. In the anthracycline group, the response rate was 40.0%. The number of patients showing poor response to the treatment was significantly larger in the patients with strongly positive PGP expression （P= 0.005）, and their prognoses were poor （P= 0.001）. in the nonanthracycline group, the response rate was 42.9%,and there was no significant relationship between the chemotherapeutic drug efficacy and the PGP or p53 protein expression. When only the data from the 11 patients treated with anthraquinone drug, mitoxantrone, were analyzed, however, the number of patients who showed poor response to treatment was significantly higher among the p53-positive patients （P= 0.012）, irrespective of the survival outcome. CONCLUSION： The chemotherapeutic efficacy with an anthracycline drug for advanced HCC can be predicted by immunohistochemical analysis of PGP expression. Similarly, immunostaining to evaluate p53 protein may be useful to predict the response in patients treated with an anthraquinone drug.

Effect of Electroacupuncture on Expression of p53 Protein in Cerebral Cortex of Rats with Global Cerebral Ischemia/Reperfusion Injury

Objective: To observe the effect of electroacupuncture (EA) on expression of p53 protein in cerebral cortex of senile rats with global cerebral ischemia/reperfusion (IR) injury and to explore its mechanism. Methods: The cerebral IR injury rat model was established referring to Pulsinelli 4-vessel occlusion method. Thirty-six SD rats were randomly and evenly divided into the control group, the IR group and the IR plus EA (IR-EA) group. The animals in the control group were subjected to electrocauterization of vertebral arteries in bilateral flank orifice alone with the general carotid arteries unoccluded. To rats in the IR-EA group, immediately and 24h, 48h, 72h after cerebral IR, EA treatment on bilateral acupoint 'Zusanli' (ST36) was applied once a day, lasting for 60 minutes. After the final treatment, all the rats were sacrificed and their brains were taken to examine p53 protein expression by the immunohistochemical method. Results: Cells with positive p53 immunoreactivity in the cerebral cortex of rats in the IR group was significantly higher than that in the control group ( P<0. 05), while that in the IR-EA group was significantly lower than that in the IR group ( P<0. 05). Conclusion: EA could remarkably reduce expression of p53 protein in the cerebral cortex of senile rats with global cerebral IR injury, which might be one of the means for EA to inhibit neuronal ap-optosis after cerebral IR injury.

EXPRESSION OF P53 PROTEIN AND PROLIFERATING CELL NUCLEAR ANTIGEN IN HUMAN GESTATION TROPHOBLASTIC DISEASE

To study the relationship between p53 protein, proliferating cell nuclear antigen (PCNA) expression and benign or malignant gestational trophoblastic disease (MGTD). Methods: The histotomic sections of 48 patients with gestational trophoblastic disease and 24 patients of normal chorionic villi were stained using immunohistochemistry. The monoclonal antibodies were used to determine p53 protein and PCNA. Results: The frequency of p53 and PCNA positive expression were significantly different among the chorionic villi of normal pregnancy, hydratidiform mole (HM) and MGTD. But neither p53 nor PCNA has any relation with the clinical staging or metastasis of MGTD. Conclusion: Both P53 and PCNA are valuable in diagnosis of human gestational trophoblastic disease.

Correlation and Expression of COX-2 and P53 Protein in Basal Cell Carcinoma of Eyelid

The correlation between the expression of COX-2 and p53 protein in basal cell carcinoma （BCC） of eyelid and apoptosis was investigated. Specimens of BCC were collected from 40 cases （aged 28-68 y） at the Department of Pathology, Renmin Hospital of Wuhan University, and Department of Pathology, Zhongnan Hospital of Wuhan University during from 1999 to 2006. Five specimens of paracancerous tissues served as control group. Immunohistochemical staining was performed to detect the expression of COX-2 and p53 in the tissues. The average absorbance （A） and the average positive area rate of COX-2 and p53 protein were measured by image analysis. The positive area rate of COX-2 and p53 protein was analyzed by linear correlation analysis. It was found that COX-2 and p53 proteins were highly expressed in BCC of eyelid, and weakly expressed in paracancerous tissues. Image analysis revealed that the expression of COX-2 and p53 proteins in BCC of eyelid was sig- nificantly higher than that in paracancerous tissues （P〈0.01）. Spearman rank correlation analysis demonstrated a positive correlation between the expression of COX-2 and p53 （r=0.113, P=0.421）. It was concluded that COX-2 can increase the expression of p53 protein, therefore suppressing apoptosis.

Effects of nanometer particles and water decoction of the Chinese medicine mixture of pinellia ternate and scorpion on P53 protein contents and apoptosis in the cerebral cortex and hippocampus of epileptic rats

BACKGROUND： Water decoction of the Chinese traditional medicine mixture of pinellia ternate and scorpion is an effective treatment for epilepsy. OBJECTIVE： To compare nanometer particles and effects of water decoction of Chinese traditional medicine mixture on P53 protein levels and apoptosis in the cerebral cortex and hippocampus of epileptic rats. DESIGN, TIME AND SETTING： This randomized, controlled molecular biology study was performed at the Key Laboratory of Child Neural Rehabilitation of Jiamusi University from October to December 2007. MATERIALS： Forty healthy male Wistar rats were used in this study. Convulsion rat models were established by intraperitoneal infusion of 35 mg/kg pentylenetetrazol, once a day, for 28 successive days. The Chinese traditional medicine mixture, comprising pinellia ternate, scorpion, centipede, bupleurum, peach pit and glycyrrhiza, was purchased from Beijing Tongrentang, China. The mixture was made into nanometer particles and water decoction. The apoptosis determination kit and P53 immunohistochemistry kit were bought from Boster, China. METHODS： Forty Wistar rats were randomly divided into four groups of ten rats per group, control, nanometer particle, water decoction and epileptic model groups. Rats in the nanometer particle and water decoction groups were respectively treated with 300 mg/kg Chinese herb nanometer particle suspension and water decoction by gavage, once a day, for 28 days. Rats in the epileptic model group were fed an equal volume of saline by gavage. Rats in the control group were only administered with the same volume of saline by gavage. MAIN OUTCOME MEASURES： Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling （TUNEL） and immunohistochemistry were used to respectively detect neuronal apoptosis and P53 protein expression in the rat brain cortex and hippocampus at 28 days following administration. RESULTS： The number of apoptotic neurons was lower in the nanometer particle and water decoction groups compared with the epileptic model group. The number of apoptotic cells and P53 protein expression in the rat cerebral cortex and hippocampus was greater in the epileptic model group compared with the control group （P 〈 0.05）, but lower in the nanometer particle and water decoction groups compared with the epileptic model group （P 〈 0.05）, and lower in the nanometer particle group compared with the water decoction group （P 〈 0.05）. CONCLUSION： Chinese traditional medicine mixture decreases neuronal apoptosis and P53 protein expression in epileptic rats. The effect of nanometer particles is significant compared with water decoction.

THE STUDY ON RELATIONSHIP BETWEEN CIGARETTE SMOKING AND THE p53 PROTEIN AND P21 PROTEIN EXPRESSION IN NON-SMALL LUNG CANCER

This paper discusses the relationship between cigarette smoking and the p53 protein and P21 protein expression by the immunohistochemical analysis in 93 cases with lung cancer in which squamous cell carcinoma accounted for 45 cases, adenocarcinoma 48 cases. The results showed that positive proportion of p53 protein expression was 74.20% (28 of 37 squamous cell carcinoma, 21 of 30 adenocarcinomas) in cigarette smoking group with lung cancers, and 38.46% (3 of 8 squamous cell carcinoma, 7 of 18 adenocarcinomas) in nonsmoking group with lung cancers. The difference was statistically significant. Odds ratio was 4.14 and confidence limits for OR was 1.42-12.52. A dose-related presents in the p53 protein expression for the smoking amount and smoking years. The positive proportion of P21 protein expression was 79.31% (21 of 28 squamous cell carcinoma, 25 of 30 adenocarcinomas) in cigarette smoking group with lung cancers, and 82.75% (10 of 11 squamous, 14 of 18 adenocarcinomas) in nonsmoking group with lung cancers, the difference was not statistically significant. But their positive proportion of P21 protein expression were very high in both groups. It was indicated that no relationship between cigarette smoking and the P21 protein expression. We suggest that the p53 gene could be a common target of tobacco-associated carcinogenesis in lung cancer.

Association between HPV16/18 Infection and Expression of P53 protein in Laryngeal Papillomas

To understand the relationship between expression of P53 protein and HPV16/18 infection in laryngeal papillomas, PCR and immunohistochemical tech-niques were used to examine the paraffin-embedded tissue samples of laryngeal pa-pillomas from 35 subjects. HPV 16/18-DNA was found in 24 cases of laryngeal papillomas (68. 8 %). Overexpression of P53 protein was detected in 19 cases (54. 3 % ). Both HPV16/18-DNA and overexpression of P53 protein were demon-strated in 12 cases of laryngeal papillomas (34. 3 % ). Our results suggest that HPV16/l8 infection and P53 gene mutation are associated with pathogenesis of la-ryngeal papillomas. The relation between HPV infection and P53 mutation in tis-sues of laryngeal papillomas remains to be clarified.

ASSOCIATION OF EXPRESSION OF P53 PROTEIN WITH CELL PROLIFERATIVE ACTIVITY AND PROGNOSIS IN COLORECTAL CANCER

The expression of p53 protein in 66 cases ofcolorectal cancer and its relationship to cell porliferativeactivity, lymph node metastasis as well as prognosis wereinvestigated by means of AB-PAP immunohistochcmicaltechnique- The results showed that 62.1/o of colorectalcancer was positive of p53 expression. The cellproliferative activity and the frequency of lymph nodemetastasis in p53- positive tumors were significantlyhigher than those in p53-negative tumors (P<0.05). Thesurvival rate in patients with p53- positive tumors wassignificantly poorer than those with p53- negative tumors(P<0.05). Our results suggest that the abnormalexpression of p53 and cell proliferation associated withmutations are involved in both human carcinogenesis andlymph node metastasis of colorectal cancer. Examinationof p53 expression is of value in understanding the degreeof malignancy, and evaluating prognosis of the disease.

Insulin Resistance in Alzheimer＇s Disease and the Role of p53 Protein

Recent evidence reveals significant metabolic changes in AD （Alzheimer＇s disease）, in which brain glucose utilization and energy production are impaired. The complex picture of metabolic disturbances in AD brains could be explained by increase of p53 protein stabilization and activation in neurons exposed to diverse metabolic stress conditions. Activated p53 helps cells to adapt to various types of metabolic stresses. Chronic stress and long lasting p53 increase lead to inhibition of glucose uptake, downregulation of glycolysis, enhancement of lipid catabolism and fatty acid β-oxidation, also promote function of tricarboxylic acid cycle and force an oxidative phosphorylation. These effects are opposite to those of insulin, thus p53 activation leads to insulin resistance of cells. However, prolonged stress conditions unveil deleterious effects of p53-evoked insulin resistance in neurons; enhancement of transcription of pro-oxidant factors, accumulation of toxic rnetabolites （e.g., cerarnide, products of advanced glycation） and ROS （reactive oxygen species）---modified cellular components, together with activation of proapoptotic genes, could finally move a suicide death program of autophagy/apoptosis in neurons. The important role of p53 in driving insulin resistance in AD brains validates attempts to inhibit p53 activity in neurons since it could promise an improvement of the disease therapy.

P53 protein expression and cell viability in irradiated peripheral blood mononuclear cells as bioindicators of radiosensitivity

Cellular radiosensitivity is directly correlated with the mechanism of DNA repair, in which p53 protein plays a major role. In this context, this study correlated cell death with p53 expression in lymphocytes irradiated in vitro with different doses of gammaradiation. For this, peripheral blood samples were collected from 10 healthy subjects. Each sample was divided in aliquots and, separately, irradiated with doses of 0,5;2 and 4 Gy. After this, peripheral blood mononuclear cells (PBMCs) were isolated and cultivated during 72 hours in 5% CO2 at 37oC without mitogen stimulation. The expression of p53 protein was evaluated by flow cytometry. In parallel, cell viability was determined by trypan blue staining. Statistical analysis was performed us-ing analysis of variance (ANOVA), differences were considered as statistically significant when p < 0.05. The results showed an increase of p53 expression with the absorbed dose, which was proportional to cell death, suggesting that p53 can be used as bioindicator of individual radiosensitivity.

Changes of p53 protein blood level in esophageal cancer patients and normal subjects from a high incidence area in Henan,China

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THE QUANTITATIVE MEASUREMENT OF BCL-2, P53 PROTEIN AND PCNA EXPRESSION IN BREAST CARCINOMA AND THEIR CORRELATION WITH PROGNOSIS

To study quantitative index of bci-2, P53, Nroliferating cell nuclear antigen (PCNA),ER and PR in breast carcinoma and their correiation and their relatiousbip with prognosis, the ex expression of bcl-2, P53 and PCNA were studied by immunohistochemical technique. The measurementof ER and PR used enzyme linked affinuity histochemical methods. The quantitative index was analyzed by image technique. All analyses were hased on 60 breast carcinomas. The results were as follows:the more bcl-2 protein, the lower histological graded the longer survival term and the highersurvival rate (P< 0. 05). The quautitative measurement of bcl-2, P53 and PCNA expression were ofvalue in evaluating the degree of differentiation and prognosis in breast carcinoma. The quantitativeand qualitative measurement or p53 protein expression showed a Ⅰwerful evidence in evaluatingprognosis of bcl-2 were more significant in evaluating poor prognosis of breast carcinoma. A relationship between bcl-2 and ER, PR showed a better value for response to endocrine therapy in breastcarcinoma patients.

Clinical significance of immunohistochemica study of P53 protein in colorectal carcinoma

AIMS To determine the clinical significance of P53 protein ex-pression in colorectal carcinoma.METHODS The expression of P53 protein in 92 colorectal car-cinomas was examined using the monoclonal antibody PAb 1801.Correlation between P53 protein expression and prognosis in col-orectal carcinoma was analyzed using log-rank test.RESULTS The frequency of P53 protein expression was 57.61%,corresponding with Dukes’ staging.Analysis of survivordata demonstrated that the survival rate of colorectal carcinomawith positive P53 protein group was lower than that of negativeP53 protein group.CONCLUSIONS We conclude that the expression of P53 pro-tein is correlated with poor prognosis in colorectal carcinoma.

Human papillomavirus and p53 protein immunoreactivity in condylomata acuminatum and squamous cell carcinoma of penis

Aim: To determine the immunoreactive pattern of human papillomavirus (HPV) antigen and p53 protein in condylo-mata acuminatum (CA) and squamous cell carcinoma (SCC) of penis. Methods: Immunohistochemistry for HPVand p53 were performed in 40 specimens of formalin fixed, paraffin embedded tissues using a polyclonal (rabbit) anti-body against HPV and a monoclonal (mouse) antibody against human p53 protein. Twenty one cases of CA and nine-teen cases of SCC were examined. Results: HPV antigen was detected in all 21 CA and 2 penile SCC. p53 proteinoverexpression was observed in 12 of 19 (63%) SCC in which 6 cases were strong positive. Five of 21 CA (24%)showed low-grade p53 protein overexpression. Conclusion; CA is related to HPV infection and some cases showp53 protein low-grade overexpression. In contrast, p53 protein overexpression is common in penile SCC, which is sel-dom related to HPV infection. (Asian J Androl 2001 Mar; 3: 75-77)

EXPRESSION OF p53 PROTEIN IN CANCERS OF SMALL INTESTINE AND ITS RELATIONSHIP TO CLINICAL COURSE AND PROGNOSIS

In order to study the relationship of p53 gene mutation with the occurrence and prognosis of cancer of small intestine, expression of p53 protein was immunohistochemically examined. The results showed that p53 protein expression was high in 75% of small intestine cancer, and positive in 21.1% of the tissues close to cancer. In 7 cases of small intestinal adenoma, only one was immunoreactive. Sixteen samples of normal tissue of the intestine didn't show expression of p53protein. The study also found that the degree of p53protein expression was significantly correlated with that of tumor cell differentiation, invasion, metastasis and prognosis.

Apoptosis and P53 protein expression after rat spinal cord injury

objective: To determine the DNA damage, neuronal and glial cells apoptosis and expression of P53 protein in the rat spinal cord after compression injury. Methods: Terminal deoxynucleotidyl transferase . mediated dUPT nick end labeling (TUNEL), DNA gel electrophoresis and immunohistochemistry techniques were used to detect DNA fragmentation in the injured rat spinal cord. Results: The apoptosic cells and P53 protein presented at 4 h after spinal cord injury with a maximum presence at 24 h in the injuried T8.9 DNA fragmentation presented typical ladder pattern on agarose gel at 24 h. Conclusion: There are lots of neuronal and glial cells apoptosis with DNA damage after SCl. The P53 protein may play an important role in induc tion of neuronal and glial cells to apoptosis.

The Value of p21^(WAF1), p53 Protein and DNA Content in Large Intestine Cancer

目的 探讨 p2 1WAF1 蛋白和 p5 3蛋白在大肠癌发生发展中的作用及其临床意义。方法 对 4 1例大肠腺癌标本及 10例正常大肠粘膜标本 ,应用免疫组织化学S -P法检测细胞周期调控因子 p2 1WAF1 蛋白和突变型 p5 3蛋白 ,应用流式细胞术检测新鲜标本DNA含量。结果 p2 1WAF1 与 p5 3表达呈负相关 ;在正常大肠粘膜、高分化腺癌、中分化腺癌、低分化腺癌中 p5 3表达率逐渐增高 ,p2 1WAF1 表达率逐渐降低 ,SPF值逐渐升高 ;Dukes分期越晚 ,p2 1WAF1 表达率越低 ,p5 3表达率越高 ,SPF越高。结论 本实验证实了 p2 1WAF1 和 p5 3表达在肿瘤发生发展中起着调控点的作用 ;p2 1WAF1 、p5

Expression and significance of P53 protein and MDM-2 protein in human gliomas

Background P53 is one of the most studied tumor suppressors in the cancer research, and over 50% of human tumors carry P53 mutations. MDM-2 is amplified and/or overexpressed in a variety of human tumors of diverse tissue origin. The aim of this study was to examine the expression of P53 protein and MDM-2 protein in gtiomas, and to investigate the relationship between the expression of the two proteins and the histopathological grades of glioma. The relationship between MDM-2 protein expression and P53 protein expression was also analyzed. Methods The expression of P53 protein and MDM-2 protein was immunohistochemically detected using monoclonal antibodies in 242 paraffin embedded tissues, including 30 normal brain tissues from patients with craniocerebral injury and 212 tissues from patients with primary glioma （grade Ⅰ-Ⅱ group： 5 cases of grade Ⅰ, 119 cases of grade Ⅱ; and grade Ⅲ-Ⅳ group： 53 cases of grade Ⅲ, and 35 cases of grade Ⅳ）. Results The P53 positive rate was significantly higher in the glioma groups than in the control group （P 〈0.0001）. The P53 positive rate was significantly higher in glioma tissues of grade Ⅲ-Ⅳ than in glioma tissues of grade Ⅰ-Ⅱ group （P=-0.001）. The MDM-2 positive rate was significantly higher in glioma groups than in the control group （P 〈0.0001）. There was no significant difference in the MDM-2 positive rate between the two glioma groups （P=0.936）. The expression of P53 protein was not related to expression of MDM-2 protein （P=-0.069） Conclusions Overexpression of P53 protein might be related to the occurrence and progression of glioma. Overexpression of MDM-2 protein may play an important role in glioma tumorigenesis, but may not be involved in glioma progression. The overexpression of MDM-2 protein was an early event in malignant transformation of glioma. MDM-2 may be a key player in glioma in its own right.

QUANTITATION OF P53 PROTEIN EXPRESSION IN GASTROINTESTINAL SMOOTH MUSCLE TUMORSCLINICOPATHOLOGICAL CORRELATION AND PROGNOSTIC SIGNIFICANCE

Quantitative analysis of P53 protein expression was performed on paraffin-embedded tissues from 55 smooth muscle tumors of the gastrointestinal tract, using immunofluo-rescence and flow cytometry. No positive expression was found in normal smooth muscle tissues of the gastrointestinal tract. Over-expression of P53 gene was found in a significantly higher proportion in leiomyosarcomas (90%) and potentially malignant smooth muscle tumors (75%) as compared to leiomyomas (14%) (P< 0.005). The quantitation of P53 expression was found to be progressively enhanced in the sequence from leiomyoma through potentially malignant smooth muscle tumor to leiomyosarcoma (P< 0.005). It was markedly over-expressed when the mitotic counts ranged from one to more than one per 10 high power fields (P< 0.005) or the mild cytologic atypia was found (P< 0.005). The five-year survival rate was significantly higher in patients with low-expression of P53 than in those with over-expression of P53 (P< 0.005). It was suggested that P53 over-expression might be associated with the transformation of leiomyoma into leiomyosarcoma and could be used as an objective parameter in distinguishing the malignant from the benign and predicting the prognosis of patients with smooth muscle rumors of the gastrointestinal tract.

DNA PLOIDY，EXPRESSION OF p53 PROTEIN AND METASTATIC BEHAVIOUR OF GASTRIC CARCINOMA

DNA ploidy of 57 gastric carcinomas with metastases(12 liver,1 adrenal,4 ovary and 48 lymph node) were measured by flow cytometry.DNA anueploidy was significantly related to liver metastases:9 out of 12 gastric carcinomas with liver metastases were anueploid(75%) as compared to 13 out of 45(28.8%) of cases without liver metastases(P<0.01);the one gastric carcinoma with adrenal metastasis was also anueploid.DNA ploidy was not related to ovarian or lymph node metastases.Another interesting finding was that all of 3 gastric carcinomas with liver metastases which showed a diploid DNA pattern,expressed p53 protein, while all of 3 carcinomas with liver metastases but no p53 protein expression were anueploid.The expression of p53 protein was not related to ovarian metastases.The results suggested that an anueploid DNA pattern and the expression of p53 protein are both objective markers valuable in predicting high risk potential of metastases to the liver,and that the combined detection of these markers can be a most useful method in the follow-up of Patients with gastric carcinoma in detecting those at high risk of developing metastases following surgical resection.Also the poorer prognosis of Patients with gastric carcinoma showing an anueploid DNA pattern may be related to the development of distant organ metastases through the blood vascular system.Furthermore,the clone of gastric carcinoma cells which accumulate p53protein or show an anueploid DNA pattern may have a causative role in the development of liver(&.adrenal) metastases.