消肿生肌汤对肛瘘术后患者肉芽组织Bax、P53、Caspase-3水平的影响

目的分析消肿生肌汤对肛瘘术后患者Bcl-2相关X蛋白(Bax)、P53、半胱氨酸蛋白酶-3(Caspase-3)水平的影响。方法选择行肛瘘术后患者80例,随机为对照组与观察组,各40例。观察组采用消肿生肌汤熏洗治疗,对照组采用常规治疗,2组均连续治疗7 d。比较2组治疗效果,治疗7 d后创面愈合情况,治疗前和治疗7 d后肉芽组织凋亡相关因子Bax、P53、Caspase-3水平和患者肛门功能。结果治疗7 d后,观察组临床有效率(95.00%,38/40)显著高于对照组(77.50%,31/40)(P<0.05);观察组创面愈合情况优于对照组(P<0.05);2组肉芽组织Bax、P53、Caspase-3水平均低于治疗前,且观察组低于对照组(P<0.05);2组肛管最长收缩时间长于治疗前,观察组长于对照组(P<0.05);肛管收缩压大于治疗前,观察组大于对照组(P<0.05);肛管静息压小于治疗前,观察组小于对照组(P<0.05)。结论消肿生肌汤熏洗能显著促进肛瘘术后患者创面愈合,降低肉芽组织凋亡相关因子Bax、P53、Caspase-3水平,改善患者肛门功能,疗效较好。

Molecular docking of DS-3032B,a mouse double minute 2 enzyme antagonist with potential for oncology treatment development

BACKGROUND It is known that p53 suppression is an important marker of poor prognosis of cancers,especially in solid tumors of the breast,lung,stomach,and esophagus;liposarcomas,glioblastomas,and leukemias.Because p53 has mouse double minute 2(MDM2)as its primary negative regulator,this molecular docking study seeks to answer the following hypotheses:Is the interaction between DS-3032B and MDM2 stable enough for this drug to be considered as a promising neoplastic inhibitor?AIM To analyze,in silico,the chemical bonds between the antagonist DS-3032B and its binding site in MDM2.METHODS For molecular docking simulations,the file containing structures of MDM2(receptor)and the drug DS-3032B(ligand)were selected.The three-dimensional structure of MDM2 was obtained from Protein Data Bank,and the one for DS-3032B was obtained from PubChem database.The location and dimensions of the Grid box was determined using AutoDock Tools software.In this case,the dimensions of the Grid encompassed the entire receptor.The ligand DS-3032B interacts with the MDM2 receptor in a physiological environment with pH 7.4;thus,to simulate more reliably,its interaction was made with the calculation for the prediction of its protonation state using the MarvinSketch®software.Both ligands,with and without the protonation,were prepared for molecular docking using the AutoDock Tools software.This software detects the torsion points of the drug and calculates the angle of the torsions.Molecular docking simulations were performed using the tools of the AutoDock platform connected to the Vina software.The analyses of the amino acid residues involved in the interactions between the receptor and the ligand as well as the twists of the ligand,atoms involved in the interactions,and type,strength,and length of the interactions were performed using the PyMol software(pymol.org/2)and Discovery Studio from BIOVIA®.RESULTS The global alignment indicated crystal structure 5SWK was more suitable for docking simulations by presenting the p53 binding site.The three-dimensional structure 5SWK for MDM2 was selected from Protein Data Bank and the three-dimensional structure of DS-3032B was selected from PubChem(Compound CID:73297272;Milademetan).After molecular docking simulations,the most stable conformer was selected for both protonated and non-protonated DS-3032B.The interaction between MDM2 and DS-3032B occurs with high affinity;no significant difference was observed in the affinity energies between the MDM2/pronated DS-3032B(-9.9 kcal/mol)and MDM2/non-protonated DS-3032B conformers(-10.0 kcal/mol).Sixteen amino acid residues of MDM2 are involved in chemical bonds with the protonated DS-3032B;these 16 residues of MDM2 belong to the p53 biding site region and provide high affinity to interaction and stability to drugprotein complex.CONCLUSION Molecular docking indicated that DS-3032B antagonist binds to the same region of the p53 binding site in the MDM2 with high affinity and stability,and this suggests therapeutic efficiency.

Tumor suppressor p53 cross-talks with TRIM family proteins

p53 is a key tumor suppressor.As a transcription factor,p53 accumulates in cells in response to various stress signals and selectively transcribes its target genes to regulate a wide variety of cellular stress responses to exert its function in tumor suppression.In addition to tumor suppression,p53 is also involved in many other physiological and pathological processes,e.g.anti-infection,immune response,development,reproduction,neurodegeneration and aging.To maintain its proper function,p53 is under tight and delicate regulation through different mechanisms,particularly the posttranslational modifications.The tripartite motif(TRIM)family proteins are a large group of proteins characterized by the RING,B-Box and coiled-coil(RBCC)domains at the N-terminus.TRIM proteins play important roles in regulation of many fundamental biological processes,including cell proliferation and death,DNA repair,transcription,and immune response.Alterations of TRIM proteins have been linked to many diseases including cancer,infectious diseases,developmental disorders,and neurodegenera-tion.Interestingly,recent studies have revealed that many TRIM proteins are involved in the regulation of p53,and at the same time,many TRIM proteins are also regulated by p53.Here,we review the cross-talk between p53 and TRIM proteins,and its impact upon cellular biological processes as well as cancer and other diseases.

Carrier-free nanoprodrug for p53-mutated tumor therapy via concurrent delivery of zinc-manganese dual ions and ROS

Human cancers typically express a high level of tumor-promoting mutant p53 protein(Mutp53)with a minimal level of tumor-suppressing wild-type p53 protein(WTp53).In this regard,inducing Mutp53 degradation while activating WTp53 is a viable strategy for precise anti-tumor therapy.Herein,a new carrier-free nanoprodrug(i.e.,Mn-ZnO_(2)nanoparticles)was developed for concurrent delivery of dual Zn-Mn ions and reactive oxygen species(ROS)within tumor to regulate the p53 protein for high anti-tumor efficacy.In response to the mild tumor acidic environment,the released Zn^(2+)and H_(2)O_(2)from Mn-ZnO_(2)NPs induced ubiquitination-mediated proteasomal degradation of Mutp53,while the liberative Mn^(2+)and increased ROS level activated the ATM-p53-Bax pathway to elevate WTp53 level.Both in vitro and in vivo results demonstrated that pH-responsive decomposition of Mn-ZnO2 NPs could effectively elevate the intracellular dual Zn-Mn ions and ROS level and subsequently generate the cytotoxic hydroxyl radical(·OH)through the Fenton-like reaction.With the integration of multiple functions(i.e.,carrier-free ion and ROS delivery,tumor accumulation,p53 protein modulation,toxic·OH generation,and pH-activated MRI contrast)in a single nanosystem,Mn-ZnO_(2)NPs demonstrate its superiority as a promising nanotherapeutics for p53-mutated tumor therapy.

Gankyrin: A potential target for drug therapy against hepatocellular carcinoma

Hepatocellular Carcinoma is a primary malignant tumor of the liver and gankyrin is an oncoprotein over-expressed in hepatocellular carcinoma. It has been found that Gankyrin protein reduces the level of p53 protein by increasing its ubiquitylation and degradation, following a MDM-2 mediated pathway. Interaction of gankyrin with MDM2 enhances the ubiquitylation of p53. Independent study of this protein molecule revealed that it is identical to the p28 subunit of the 26S proteasome, having seven similar alpha helical ankyrin repeats. Gankyrin also binds to the Tumor Suppressor Protein (TSP) Retinoblastoma (RB), thereby accelerating its phosphorylation and proteasomal degradation. Blocking the expression of Gankyrin with MDM2 in cases of Hepatocellular Carcinoma (HCC) promoted apoptosis in cancer cells. Hence, Gankyrin can be used as a potential target for drug therapy against Hepatocellular Carcinoma.

Natural products targeting the p53-MDM2 pathway and mutant p53: Recent advances and implications in cancer medicine

The p53 tumor suppressor plays a major role in controlling the initiation and development of cancer by regulating cell cycle arrest,apoptosis,senescence,and DNA repair.The MDM2 oncogene is a major negative regulator of p53 that inhibits the activity of p53 and reduces its protein stability.MDM2,p53,and the p53-MDM2 pathway represent welldocumented targets for preventing and/or treating cancer.Natural products,especially those from medicinal and food plants,are a rich source for the discovery and development of novel therapeutic and preventive agents against human cancers.Many natural product-derived MDM2 inhibitors have shown potent efficacy against various human cancers.In contrast to synthetic small-molecule MDM2 inhibitors,the majority of which have been designed to inhibit MDM2-p53 binding and activate p53,many natural product inhibitors directly decrease MDM2 expression and/or MDM2 stability,exerting their anticancer activity in both p53-dependent and p53-independent manners.More recently,several natural products have been reported to target mutant p53 in cancer.Therefore,identification of natural products targeting MDM2,mutant p53,and the p53-MDM2 pathway can provide a promising strategy for the development of novel cancer chemopreventive and chemotherapeutic agents.In this review,we focus our discussion on the recent advances in the discovery and development of anticancer natural products that target the p53-MDM2 pathway,emphasizing several emerging issues,such as the efficacy,mechanism of action,and specificity of these natural products.

Effects of HBV X gene and arsenic trioxide on the expression of p53 in cultured HepG2 cells

Background Hepatitis B virus （HBV） X protein （HBx） and p53 could mutually down-regulate at transcriptional level and HBx could bind with p53 protein within its transactivation domain and inhibit the function of p53 protein. In recent years, effects of arsenic trioxide （As2O3） on the expression of p53 protein have been widely studied, while little is known about the activity of p53 protein. This study was undertaken to delineate the effect of HBV X gene and AS203 on p53 protein expression （level and activity） in HepG2 cells by small hairpin RNA （shRNA）-mediated RNA interference （RNAi） technique. Methods Cell line HepG2 and cells with stable expression of HBV X gene （HepG2-X） were treated with 2 pmol/L AS203, with corresponding untreated cells serving as controls. Cell lysates and nuclear extracts were extracted. Total level and the relative activity of p53 protein were detected by modified enzyme-linked immunosorbent assay （ELISA）. HBV X gene sequence-specific shRNA expression vector （pXi-1 and pXi-2） and sequence-unrelated control （pXi-3） were transfected into HepG2-X. Single cell clone with stable expression of shRNA was selected and exposed to propagating culture. The effect of As2O3 on p53 protein expression and activity was re-observed. Results Total p53 protein level was up-regulated and its relative activity ratio was enhanced by As2O3 in HepG2 and HepG2-X cells. The total p53 protein level induced by As2O3 was up-regulated by HBV X gene expression, while its relative activity was significantly suppressed. The suppression was removed after HBV X gene expression was repressed by shRNA. Conclusions As2O3 up-regulates p53 protein expression and enhance its activity. HBV X up-regulates As2O3 induced-p53 protein expression while suppresses its activity.

USH2A mutation and specific driver mutation subtypes are associated with clinical efficacy of immune checkpoint inhibitors in lung cancer

This study aimed to identify subtypes of genomic variants associated with the efficacy of immune checkpoint inhibitors(ICIs)by conducting systematic literature search in electronic databases up to May 31,2021.The main outcomes including overall survival(OS),progression-free survival(PFS),objective response rate(ORR),and durable clinical benefit(DCB)were correlated with tumor genomic features.A total of 1546 lung cancer patients with available genomic variation data were included from 14 studies.The Kirsten rat sarcoma viral oncogene homolog G12C(KRAS^(G12C))mutation combined with tumor protein P53(TP53)mutation revealed the promising efficacy of ICI therapy in these patients.Furthermore,patients with epidermal growth factor receptor(EGFR)classical activating mutations(including EGFRL858Rand EGFRΔ19)exhibited worse outcomes to ICIs in OS(adjusted hazard ratio(HR),1.40;95%confidence interval(CI),1.01-1.95;P=0.0411)and PFS(adjusted HR,1.98;95%CI,1.49-2.63;P<0.0001),while classical activating mutations with EGFR^(T790)Mshowed no difference compared to classical activating mutations without EGFR^(T790)Min OS(adjusted HR,0.96;95%CI,0.48-1.94;P=0.9157)or PFS(adjusted HR,0.72;95%CI,0.39-1.35;P=0.3050).Of note,for patients harboring the Usher syndrome type-2A(USH2A)missense mutation,correspondingly better outcomes were observed in OS(adjusted HR,0.52;95%CI,0.32-0.82;P=0.0077),PFS(adjusted HR,0.51;95%CI,0.38-0.69;P<0.0001),DCB(adjusted odds ratio(OR),4.74;95%CI,2.75-8.17;P<0.0001),and ORR(adjusted OR,3.45;95%CI,1.88-6.33;P<0.0001).Our findings indicated that,USH2A missense mutations and the KRAS^(G12C)mutation combined with TP53 mutation were associated with better efficacy and survival outcomes,but EGFR classical mutations irrespective of combination with EGFR^(T790)Mshowed the opposite role in the ICI therapy among lung cancer patients.Our findings might guide the selection of precise targets for effective immunotherapy in the clinic.

PLK1和P53蛋白在卵巢上皮性癌中的表达及其意义

2目的检测POLO样蛋白激酶1（PLK1）和P53蛋白在卵巢上皮性癌中的表达，探讨其与患者预后的关系。方法采用免疫组织化学SP法检测20例正常卵巢组织、19例卵巢良性上皮性肿瘤组织和52例卵巢上皮性癌组织中PLK1和P53蛋白的表达情况，分析其与卵巢癌临床病理指标的关系及二者在卵巢癌组织中表达的相关性，单因素和多因素Logistic回归分析影响卵巢癌患者预后的危险因素，Kaplan-Meier法分析PLK1和P53表达与卵巢癌患者预后的关系。结果 PLK1和P53在卵巢上皮性癌组织中的表达分别为38.5%和67.3%，显著高于良性肿瘤和正常卵巢组织（P＜0.05）。在卵巢上皮性癌中，PLK1和P53异常高表达与临床分期和组织分化相关，临床分期越晚、组织分化越差的癌组织中PLK1和P53蛋白表达越高（P＜0.05）； PLK1蛋白的表达与P53蛋白的表达呈负相关（r＝-0.629，P＝0.000）。单因素Logistic回归分析显示PLK1、P53、临床分期、组织分化和淋巴结转移是影响卵巢癌患者预后的因素，多因素Logistic回归分析显示仅PLK1是影响卵巢癌患者预后的独立因素（OR＝2.288，P＝0.025，95% CI：0.105～50.050）。与其他患者相比，PLK1表达阳性同时P53表达阳性的卵巢癌患者，生存期最短（χ2＝17.246，P＝0.037）。结论 PLK1和P53共同参与了卵巢癌的发生发展，PLK1可作为判断卵巢癌患者预后的标志物。

几种多药耐药相关因子在肺癌中的表达及临床病理意义

目的:探讨原发性肺癌中几种与多药耐药相关因子的表达及与临床病理的相关性。方法:采用免疫组化方法对术前未经化疗、术后病理诊断明确的133例肺癌组织标本和15例作为对照的癌旁肺组织(距肿瘤边缘5cm),进行了P-gp、MRP、LRP、p53及c-erbB-2蛋白表达的检测。结果:(1)133例肺癌组织中,P-gp、MRP、LRP、p53及c-erbB-2的阳性率分别为72.2%(96/133)、40.5%(51/126)、54.1%(72/126)、59.4%(79/126)和45.9%(61/133),除P-gp外均显著高于正常肺组织中的表达水平(P<0.05);且p53表达与P-gp及MRP表达之间(r=0.179及0.185,P=0.039及0.041),P-gp与LRP之间(r=0.264,P=0.002),LRP与MRP之间(r=0.309,P<0.001),以及c-erbB-2表达分别与MRP、LRP及p53表达之间(r分别为:0.350、0.421、0.541和0.354,P值均<0.001)均有相关性。(2)P-gp、MRP、LRP和c-erbB-2蛋白表达在不同病理类型之间具有统计学差异,而LRP蛋白表达在不同病理分化程度之间具有统计学差异(P<0.001),P-gp在鳞癌不同分化程度间表达有统计学差异(P=0.049)。(3)针对病理类型及亚型:①神经内分泌癌与非神经内分泌癌相比,后者的LRP和c-erbB-2蛋白表达水平较前者高(P=0.005和0.041);②未发现细支气管肺泡癌与其他类型腺癌间在MDR相关因子的蛋白表达水平上有统计学差异。结论:这些多药耐药相关因子可能均在肺癌的原发性多药耐药中起不同程度的作用,且在多药耐药形成以及肿瘤进展的过程中它们之间有某种程度的关联。

热休克蛋白B1基因、肿瘤抑制蛋白p53基因多态性与焦炉作业工人染色体损伤的关联及其交互作用分析

目的：探讨热休克蛋白B1基因（HSPB1）启动子区多态性位点rs2868371和肿瘤抑制蛋白P53基因（TP53）基因多态性位点rs1042522与焦炉作业工人染色体损伤的关联及其交互作用。方法于2010年9—10月，选取武汉市某焦化厂1333名男性焦化厂工人作为调查对象，以其中在辅助车间、炉底、炉侧和炉顶等场所工作的工人作为暴露组，共949名；在办公室场所工作的行政、医务人员作为对照组，共384名。收集调查对象的人口学信息并采集外周静脉血5 ml，分别运用ELISA和胞质分裂阻滞试验测定其血浆苯并[a]芘二氢二醇环氧化物-白蛋白（BPDE-白蛋白）加合物的浓度和外周血淋巴细胞微核率，采用TaqMan探针技术和ABI Prism 7900HT型荧光定量PCR仪进行SNP分型检测，并计算频率比（FR）及其95%CI值。结果暴露组中，携带rs2868371 GG基因型者外周血淋巴细胞微核率为（3.52±2.67）‰，均低于携带GC、CC及GC＋CC基因型者[分别为（3.88±2.88）‰、（4.00±2.66）‰和（3.91±2.83）‰]（FR值分别为1.10、1.13、1.11，95%CI值分别为1.02~1.19、1.02~1.25、1.03~1.19），且rs2868371 C有明显的等位基因剂量-效应关系（P趋势=0.006）；携带rs1042522 CC基因型者微核率为（3.95±3.06）‰，均高于携带CG以及CG＋GG基因型者[分别为（3.63±2.61）‰、（3.66±2.61）‰]（FR值分别为0.87、0.90，95%CI值分别为0.83~0.96、0.84~0.97）。HSPB1 rs2868371和TP53 rs1042522对外周血淋巴细胞微核率水平存在交互作用（P交互=0.001）。年龄〉40岁（FR=1.07，95%CI：1.01~1.12）、工龄〉20年（FR=1.08，95%CI：1.02~1.14）、BMI≤24 kg/m2（FR=1.07，95%CI：1.01~1.13）、饮酒（FR=1.09，95%CI：1.02~1.16）以及高BPDE-白蛋白水平（FR=1.07，95%CI：1.01~1.13）者HSPB1 rs2868371 C等位基因与微核率的增加均具有等位基因剂量-效应关系（P趋势分别为0.023、0.013、0.029和0.020）；年龄〉40岁（FR=0.94，95%CI：0.89~0.99）、BMI≤24 kg/m2（FR=0.94，95%CI：0.88~0.99）以及饮酒（FR=0.94，95%CI：0.88~1.00）者TP53 rs1042522 G等位基因与微核率的降低均具有等位基因剂量-效应关系（P趋势分别为0.031、0.023和0.038）。HSPB1 rs2868371与年龄和工龄在影响外周血淋巴细胞微核率上存在交互作用（P交互值分别为0.030和0.013）。结论 HSPB1 rs2868371 C等位基因与焦炉工染色体损伤水平增加有关联，TP53 rs1042522 G等位基因与焦炉工染色体损伤水平降低有关联，且上述两个SNP位点在影响焦炉工的染色体损伤水平上具有交互作用。

Overexpression of Bcl-2 partly inhibits apoptosis of human cervical cancer SiHa cells induced by arsenic trioxide

OBJECTIVE: To study the biological effect of arsenic trioxide (As2O3) on human cervical cancer SiHa cells and SiHa cells overexpressing bcl-2 gene. METHODS: SiHa cells with overexpression of Bcl-2 (SiHa-Bcl2 cells) were established by transfecting SiHa cells with Bcl-2 expression vector. The sensitivities of SiHa and SiHa-Bcl2 cells to As2O3 were determined using MTT (Thiazolyl blue) reduction and colony forming ability assay, morphological analysis, flow cytometric analysis, DNA agarose gel electrophoresis, in situ cell death detection (TUNEL), Northern blot, RT-PCR and Western blot. RESULTS: As2O3 inhibited the growth of SiHa cells and induced G2/M arrest and apoptosis of the cells. RT-PCR and Western blot analysis revealed that As2O3 induced SiHa cell apoptosis possibly via inhibiting the expression of HPV16 E7 and decreasing the expression of c-myc. However, we found that SiHa-Bcl2 cells partly resisted As2O3 induced apoptosis, which might be related to the prevention of the down-regulation of HPV16 E7 and c-myc gene expression. Nevertheless, As2O3 at a high concentration could still induce apoptosis of SiHa-Bcl2 cells mainly via decreasing Bcl-2 expression and slightly inhibiting viral gene expression. CONCLUSION: As2O3 is an inducer of the apoptosis of human cervical carcinoma cells and the cells overexpressing Bcl-2 can partly resist As2O3 induced apoptosis, but the exact mechanism is unclear.

Expression of Syk in non-small cell lung cancer and its relationship with clinicopathological parameters

This study aims to research the expression of spleen tyrosine kinase(Syk)in non-small cell lung cancer(NSCLC)and the relationship between Syk and clinico-pathologic factors and p53.Immunohistochemistry was applied to detect the expression of Syk and p53 protein in 39 cases of NSCLC(23 cases of lung squamous cell can-cer,16 cases of lung adenocarcinoma)and tumor-sur-rounding normal lung tissues.The positive rate of Syk was 46.15%(18/39)and 100%(39/39)in NSCLC and tumor-surrounding normal lung tissues,respectively.The expres-sion level of Syk in NSCLC was significantly lower than that in tumor-surrounding normal lung tissues(P=0.000).The Syk expression was positively correlated with the p53 expression in NSCLC specimens(P=0.025).There was no significant association between Syk expression and lymph node metastasis,differentiation degree,tumor size and tumor node metastasis(TNM).The present study demonstrated that Syk was aberrantly expressed in the NSCLC and might have a significant impact on tumor growth and progression.

Extracellular and intracellular functions of coiled-coil domain containing 3

Coiled-coil domain containing 3(CCDC3,also called Favine)is a highly conserved protein initially identified as a protein secreted from adipocytes and endothelial cells in the vascular system with endocrine-like functions.Recently,CCDC3 was also found to function as a nuclear tumor suppressor in breast cancers.Although it is still understudied,CCDC3,since its discovery,has been shown to play multiple roles in lipid metabolism,fatty liver,abdominal obesity,anti-inflammation,atherosclerosis,and cancer.This essay is thus composed to offer an overview of these extracellular endocrine-like and intracellular(nuclear)functions of CCDC3.We also discuss the possible underlying cellular and molecular mechanisms of CCDC3,the implications for clinical translation,and the remaining puzzles about this special molecule.

大肠肿瘤细胞凋亡及相关基因表达的研究

为了探讨Bcl-2和P53蛋白在大肠肿瘤中的表达及与细胞凋亡的关系,用免疫组化方法观察了45例大肠腺瘤和61例大肠癌中Bcl-2和P53蛋白的表达.正常大肠粘膜中Bcl-2和P53均未见表达,而大肠腺癌及大肠癌阳性率均较正常明显增加(P<0.01).大肠腺瘤P53表达随腺瘤大小增加而增加,其中≥20mm组阳性率(77.78%)显著高于<10mm组(35.00%,P<0.05).P53蛋白阳性率也随不典型增生程度增加而增高.P53表达与大肠癌分化程度及Dukes分期有关.大肠癌细胞凋亡指数与Bcl-2阳性表达呈负相关.大肠腺癌中Bcl-2和P53蛋白的表达也呈负相关.Bcl-2蛋白对大肠癌前病变-脉瘤的增殖有一定表达意义,P53在大肠腺瘤癌变和大肠癌进展中起重要作用.它们也是参与细胞凋亡的良好指标.