Effect of HCV NS_3 protein on P53 protein expression in hepatocarcinogenesis

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Function of apoptosis and expression of the proteins Bcl-2,p53 and C-myc in the development of gastric cancer

INTRODUCTIONIn China ,the incidence and mortality of gastric cancer rank the second among all cancers. Recent development of cancer [1-20].The aim of this study was investigat the insight of apoptosis and bcl-2, p53 and C-myc protein expression in the development of gastric cancer .

Effect of Boschniakia rossica on expression of GSTP,p53 and p21^(ras)proteins in early stage of chemical hepatocarcinogenesis and its anti-inflammatory activities in rats

AIM To investigate the effect of Boschniakiarossica（BR）extract on expression of GST-P,p53 and p21rasproteins in early stage of chemicalhepatocarcinogenesis in rats and its anti-inflammatory activities.METHODS The expression of tumor marker-placental form glutathione S-transferase（GST-P）,p53 and p21rasproteins were investigated byimmunohistochemical techniques and ABCmethod.Anti-inflammatory activities of BR werestudied by xylene and croton oil-induced mouseear edema,carrageenin,histamine and hotscald-induced rat pow edema,adjuvant-inducedrat arthritis and cotton pellet-induced mousegranuloma formation methods.RESULTS The 500 mg/kg of BR-H2O extractfractionated from BR-Methanol extract hadinhibitory effect on the formation of DEN-inducedGST-P-positive foci in rat liver（GST-P stainingwas 78% positive in DEN+AAF group vs 20%positive in DEN+AAF+BR group,P【0.05）andthe expression of mutant p53 and p21rasproteinwas lower than that of hepatic preneoplasticlesions（33% and 22% positive respectively inDEN+AAF group vs negative in DEN+AAF+BRgroup）.Both CH2Cl2 and H2O extracts from BRhad anti-inflamatory effect in xylene and crotonoil-induced mouse ear edema（inhibitory rateswere 26%-29% and 35%-59%,respectively）. BR-H2O extract exhibited inhibitory effect incarrageenin,histamine and hot scald-inducedhind paw edema and adjuvant-induced arthritis inrats and cotton pellet-induced granulomaformation in mice.CONCLUSION BR extract exhibited inhibitory effect on formation of preneoplastic hepatic foci in early stage of rat chemical hepato-carcinogenesis. Both CH2CI2 and H2O extracts from BR exerted anti-inflammatory effect in rats and mice.

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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Prognostic values of apoptosis-stimulating P53-binding protein 1 and 2 and their relationships with clinical characteristics of esophageal squamous cell carcinoma patients:a retrospective study

Background: Esophageal squamous cell carcinoma(ESCC) is a leading cause of cancer?related death, and new prognostic biomarkers are urgently needed. Apoptosis?stimulating P53?binding protein 1(ASPP1) and 2(ASPP2) have been reported to play important roles in the development, progression, metastasis, and prognosis of cancers, but their roles in ESCC have not been elucidated. In this study, we examined the expression of ASPP1 and ASPP2 in ESCC to evaluate their prognostic values.Methods: The protein expression of ASPP1, ASPP2, and P53 in 175 specimens of ESCC was detected using immuno?histochemical staining; their expression in cancerous and noncancerous tissues was scored according to the stain?ing intensity and the percentage of stained cells. The associations of ASPP1, ASPP2, and P53 with clinicopathologic parameters, overall survival(OS), and disease?free survival(DFS) were analyzed.Results: The protein expression levels of ASPP2 and P53 were significantly higher in cancerous tissues than in paired noncancerous tissues(P < 0.001), whereas the expression levels of ASPP1 in the two groups were similar. In ESCCs, ASPP1 expression was significantly associated with histological differentiation(P = 0.002) and invasive depth(P = 0.014); ASPP2 expression was associated with age(P = 0.029) and histological differentiation(P < 0.001); and P53 expression was associated with age(P and P53 expression. Survival an= 0.021) and tumor size(P alysis revealed that high AS= 0.040). No correlations were found between ASPP1, ASPP2,PP2 expression was significantly associated with increased 5?year OS(P = 0.001) and DFS rates(P ate of ESCC patients(= 0.010) and that high P53 expression was significantly associated with a reduced 5?year DFS rP atio(HR): 0.541, 9= 0.015). Multivariate Cox analysis indicated that ASPP2 was an inde?pendent predictor of OS [hazard r5% confidence interval(CI) 0.363–0.804] and DFS(HR: 0.599, 95% CI 0.404–0.888) of ESCC patients and that P53 was an independent predictor of DFS(HR: 2.161, 95% CI 1.100–4.245).Conclusions: ASPP1 might be involved in the progression of ESCC, and ASPP2 was a potential prognostic biomarker of ESCC and should be evaluated in future studies.

Role of microRNA-21 in uveal melanoma cell invasion and metastasis by regulating p53 and its downstream protein

AIM： To reveal the insight mechanism of liver metastasis in uveal melanoma, we investigated cell functions of microRNA-21 in three different uveal melanoma cell lines and analyze the relationship of target gene p53 and its downstream targets which been found significant expression in our previous study.METHODS： Quantitative real-time polymerase chain reaction （qRT-PCR） was used to detect microRNA-21 expression in normal uveal tissue and uveal melanoma cell lines. Lenti-virus expression system was used to construct OCM-1, MuM-2B and M619 cell line with stable overexpression and inhibition of microRNA-21. In vitro cell function tests such as cell proliferation, cell apoptosis, cell circle and abilities of migration and invasion were examined by MTT, BrdU assay, flow cytometry, transwell assay and Matrigel invasion assay respectively. The target gene was predicted by bioinformatics and confirmed by using a dual luciferase reporter assay. The expression of p53 and its suspected downstream targets LIM and SH3 protein 1 （LASP1） and Glutathione S Transferase pi （GST-Pi） were determined by qRT-PCR in mRNA level and western blotting analysis in protein level. Finally, the effect of microRNA-21 in a xenograft tumor model was assessed in four-week-old BALB/c nude mice. RESULTS： Compared to normal uveal melanoma, expressions of microRNA-21 were significantly higher in uveal melanoma cell lines. Overexpression of microRNA-21 promoted proliferation, migration, and invasion of OCM-1, M619 and MuM-2B cells, while inhibition of microRNA-21 reveal opposite effects. Wild type p53 was identified as a target gene of microRNA-21-3p, and proved by dual luciferase reporter assay. Up-regulated microRNA-21 inhibited the expression of wild type p53 gene, and the increased expression of LASP1 in mRNA level and protein level, while down-regulated microRNA-21 presented opposite way. However, GST-pi showed the potential pattern as expected, but relative mRNA level showed no statistically significant difference in OCM-1 cells. Furthermore, the mRNA expression of GST-pi was decreased in microRNA-21 overexpressing MuM-2B, and increased in M619 cells with inhibition of microRNA-21. In vivo, inhibition of microRNA-21 reduced tumor growth with statistically significant difference.CONCLUSION： These findings provide novel insight into molecular etiology of microRNA-21 in uveal melanoma cell lines, and suggest that microRNA-21 might be a potential candidate for the diagnosis and prognostic factor of human uveal melanoma in future.

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)

p53 MUTATIONS AND PROTEIN OVEREXPRESSION IN PRIMARY COLORECTAL CANCER AND ITS LIVER METASTASIS

Objective: To compare p53 status in primary and hepatic metastatic colorectal cancer in 34 patients. Methods: p53 gene status (exons 5–9) was examined by PCR, denaturing gradient gel electrophoresis (DGGE) and automated sequencing. P53 protein was detected by immunohistochemistry using monoclonal antibody DO-7. Results: p53 mutations were found in exons 5 through 9 in 21 of 34 patients (61.8%). Among them, 5 patients had mutation in liver metastasis but not in their primary tumors while in the other patients the same mutations were found in both primary and metastatic colorectal cancers. In no patients was p53 mutation exclusively found in the primary colorectal tumors. Moreover, additional mutation was detected in the metastatic lesions in two cases. Of the 37 mutations within the exons examined, 73% was missense mutation and 16% was nonsense mutation. There were 4 microinsertions. p53 protein was overexpressed in both primary and metastatic colorectal cancers with p53 gene mutations. The presence of p53 mutation significantly correlated with p53 protein accumulation (r=0.96,P<0.001). However, in 4 patients with p53 nonsense mutation, immunohistochemical staining was negative. In three patients who showed no p53 mutation of the primary tumor, p53 protein was consistently overexpressed. Conclusion: In colorectal cancers, p53 gene mutation usually appears first in the primary tumor and maintains as such but is more prominent when metastasized to the liver. However, p53 gene mutation may occur only after being metastasized. Although p53 gene mutation and p53 protein overexpression correlate with each other, either parameter examined alone may lead to false positive or negative results.

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.

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.

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.

贲门癌组织P53、P21和CD_(44V6)蛋白的表达及其意义

目的 探讨P53 、P2 1和CD44V6蛋白的表达情况与贲门癌生物学行为的关系。方法 采用免疫组化法对 4 1例贲门癌组织进行P53 、P2 1和CD44V6蛋白的测定。结果 贲门癌组织中P53 、P2 1和CD44V6蛋白的阳性率分别为 75 6 %、5 3 6 %和 4 1 4 % ,其表达与患者性别、年龄、临床分期和病理类型无明显关系。CD44V6蛋白阳性表达与肿瘤浸润深度有关 (P <0 0 5 ) ;P53 蛋白表达与P2 1蛋白表达呈正相关 ,但无统计学意义。P53 、P2 1和CD44V6蛋白阳性者预后比其阴性者差。结论 P53 、P2 1和CD44V6蛋白的测定可作为判断贲门癌患者预后的参考指标 。

p53、bcl-2和CD44v6蛋白表达与乳腺癌转移的相关性研究

目的探讨乳腺癌及癌旁正常乳腺组织中p53、bcl2和CD44v6蛋白的表达及意义。方法采用免疫组化法检测96例乳腺癌及其癌旁正常乳腺组织中p53、bcl2和CD44v6蛋白的表达,并分析其与淋巴结转移和cerbB2表达状态的相关性,从而评价这些指标在预测乳腺癌转移方面的价值。结果正常乳腺组织中p53蛋白为阴性,在乳腺癌中阳性表达率为65.63%;随着组织学分级的增高,阳性率逐渐增高;淋巴结转移组阳性表达率明显高于无淋巴结转移组,并与cerbB2表达状态呈正相关。乳腺癌组织中bcl2阳性表达率明显低于周围正常乳腺组织,随着组织学分级的增高,阳性率逐渐降低,淋巴结转移组中的表达率明显低于淋巴结非转移组,与cerbB2的表达呈负相关。CD44v6在乳腺癌组织中的阳性表达率明显高于正常乳腺组织,随着组织学分期的增加,CD44v6的阳性表达率亦增高,但差异无显著性,淋巴结转移组的阳性率略高于无淋巴结转移组,差异也无显著性,与cerbB2表达无相关性。结论p53和bcl2蛋白可作为预测乳腺癌转移的指标,CD44v6的阳性表达可能与乳腺癌的发生、进展有一定关系,但尚不能把它作为预测乳腺癌转移的稳定的生物学指标。

粘附分子CD44v6和p53基因在乳腺癌组织中的表达及其与DNA倍体的关系

背景与目的:细胞表面粘附分子CD44的变构体CD44v6在肿瘤转移中起重要作用,但对其在乳腺癌中的表达研究结果差异较大。p53基因突变是最常见的与人类肿瘤发生相关的基因异常。本研究探讨CD44v6和p53基因及DNA倍体在乳腺癌中的表达意义与相互关系。方法:采用流式细胞光度术(FCM)定量分析了52例原发性乳腺癌组织、相应癌旁组织和18例乳腺良性肿瘤中P53和CD44v6及DNA倍体的变化状况。结果:乳腺癌组织中P53、CD44v6蛋白表达及S期比例(SPF)和增殖指数(PI)显著高于相应癌旁组织、乳腺良性肿瘤和正常乳腺组织(P<0.05)。P53、CD44v6、SPF、PI在不同年龄、不同病理类型、不同肿瘤大小和临床分期间的表达差异均无显著意义(P>0.05)。有淋巴结转移组患者P53、CD44v6表达和SPF、PI值明显高于无淋巴结转移组患者(P<0.05)。52例乳腺癌中异倍体23例(44.23%),20例发生转移;二倍体29例,17例发生转移,异倍体乳腺癌患者淋巴结转移率明显高于二倍体乳腺癌患者(P<0.05),且异倍体乳腺癌患者P53蛋白表达明显高于二倍体乳腺癌患者(P<0.05),而CD44v6表达在两者间未见显著差异(P>0.05)。相关分析显示,癌组织P53蛋白表达与SPF和PI具显著正相关性(P<0.01),而CD44V6与SPF和PI均无关(P>0.05)。结论:P53、CD44v6蛋白表达和DNA含量在乳腺癌?

大肠癌组织中EGFR、c-erbB-2、CD_(44)v6和p53异常表达与3年生存率分析

目的:同时检测癌基因EGFR、c-erbB-2、CD44v6和p53在大肠癌组织中的异常表达,分析不同表达水平患者组的3年生存率差异。方法:对63例存档大肠癌石蜡组织标本进行重新切片,分别采用EGFR、c-erbB-2、CD44v6和p53单克隆抗体进行免疫组化染色(SABC法)。所有病例均随访3年以上。结果:全部63例大肠癌中除4例(6.3%)无任何1种癌基因蛋白表达外,11例(17.5%)表达1种癌基因蛋白的患者3年生存率88.8%;19例(30.2%)表达2种癌基因蛋白的患者3年生存率70.5%;15例(23.8%)表达3种癌基因蛋白的患者3年生存率57.8%;14例(22.2%)表达全部4种癌基因蛋白的患者3年生存率22.2%,与前3组患者的3年生存率比较均有显著性差异(P<0.01)。结论:同时检测大肠癌组织4种癌基因蛋白异常表达可准确判断患者不良预后。

大肠癌中CD44V6、p53、PCNA的表达及意义

目的 探讨 CD44 V6、p5 3、PCNA在大肠癌中的表达与病理及生理学行为之间的关系。方法 采用免疫组化S- P法检测 CD44 V6、p5 3、PCNA在 5 1例大肠癌中的表达。结果 5 1例大肠癌 CD44 V6阳性表达率 41.18% (2 1/ 5 1)。CD44 V6在低分化大肠癌的检出率为 5 3.85 % (7/ 13)明显高于高分化大肠癌 36 .84% (7/ 19) (P<0 .0 5 ) ,在浆膜外的阳性表达率 5 3.38% (8/ 15 )明显高于肌层 33.33% (2 / 6 ) (P<0 .0 5 ) ,有淋巴结转移者阳性表达率为 6 3.16 % (12 / 19)明显高于无淋巴结转移者 31.2 5 % (10 / 32 ) (P<0 .0 5 )。p5 3在 5 1例大肠癌中阳性检出率为 5 0 .5 9% (2 6 / 5 1) ,其表达与大肠癌的组织类型、分化程度和淋巴结转移未见明显相关。而与浸润深度呈正相关 ,p5 3在浆膜外层的阳性表达率为5 3.33% (8/ 15 )明显高于肌层 33.33% (2 / 6 ) (P<0 .0 5 )。 PC-NA的表达与病理分级、浸润深度和淋巴结转移未见显著相关性 (P>0 .0 5 )。结论 CD44 V6的表达与大肠癌的临床病理生理学行为密切相关 ,而 p5 3与大肠癌浸润深度关系密切。因此 CD44 V6和 p5 3蛋白可为一个准确预测大肠癌预后的生物学指标。而

p53与CD44V6基因表达与肾癌转移相关性研究

目的 探讨 p5 3与 CD44 V6基因的表达与肾癌转移相关性。方法 采用 L SAB法对 35例肾癌 (16例转移 ) p5 3与 CD44 V 6基因的表达进行了免疫组织化学检测 ,并结合 CK- H,CK- L ,Vimentin的表达情况进行综合分析。结果 p5 3在肾细胞癌转移组表达率 (6 0 .0 % )与无转移组中的表达率 (30 .0 % )差异有显著性 (P<0 .0 5 ) ,其表达率随肾细胞癌组织学分级和临床分期的增加呈增高趋势 , 级 (44 .4% )与 ～ 级 (10 0 % ) , 期 (2 0 .0 % )与 、 ～ 期 (5 0 .0 % ,6 2 .5 % )差异有显著性 (P<0 .0 1,P<0 .0 5 ) ,CD44 V6基因的表达与肾癌组织学分级呈正相关。CK- L的表达情况与 p5 3类似。结论 p5 3、CD44 V6基因、CK- L等的表达与肾癌的发生、分化、转移等因素密切相关 ,对肾细胞癌的预后判断有一定的参考价值。

结直肠癌腹腔转移与CK20 mRNA、CD44v6和P53表达及意义

目的 研究结直肠癌腹腔转移与CK2 0mRNA、CD44v6和P5 3的表达及意义。方法 采用逆转录 聚合酶链反应 (RT PCR)技术 ,对 5 0例结直肠癌进行腹腔渗液CK2 0mRNA测定 ,并且对癌组织行免疫组化检测CD44v6和P5 3。结果 CK2 0mRNA阳性表达与癌组织中P5 3和CD44v6之间存在显著性差异。结论 检测腹腔渗液中的CK2 0mRNA并结合癌组织中P5 3和CD44v6的检测 ,对于预测结直肠癌腹腔转移和指导术后治疗具有重要参考价值。

p53 CD44V6及nm23/NDPK在非小细胞肺癌中表达的相关性分析

目的:探讨非小细胞肺癌(NSCLC)中p53、CD44V6及nm23/NDPK表达的相关性及预后意义。方法:采用免疫组织化学SP法对69例NSCLC组织中p53、CD44V6及nm23/NDPK进行检测。结果:p53、CD44V6及nm23/NDPK的表达与NSCLC患者的临床分期(P<0.05,P<0.01)、淋巴结转移(P<0.05,P<0.01)及生存率(P<0.05)密切相关。p53、CD44V6表达与nm23/NDPK在NSCLC中的表达呈明显负相关(P<0.01)。结论:联合检测NSCLC中p53、CD44V6及nm23/NDPK的表达对于判断预后和指导治疗可能具有实用价值。