曲古抑菌素A通过p53转录激活途径诱导尤文肉瘤细胞凋亡

目的：探讨组蛋白去乙酰化酶抑制剂曲古抑菌素A（trichostatin A,TSA）诱导尤文肉瘤细胞株WE-68和VH-64凋亡及作用机制.方法：四甲基偶氮唑蓝法（MTT法）测定细胞增殖抑制率.流式细胞计数法测量TSA给药后细胞周期中sub-G1含量的变化.免疫印迹法（Western-blot）检测细胞中活化型多聚ADP核糖多聚酶（cleaved-PARP）,p53-lys382残基乙酰化和p53蛋白总量的表达.实时定量PCR和siRNA转染技术测定p53多个下游基因的mRNA水平改变和p53表达下调对TSA诱导凋亡的影响.结果：TSA抑制了尤文肉瘤细胞的增殖,诱导细胞周期中sub-G1含量和凋亡终产物cleaved-PARP蛋白表达的增加.TSA给药后,p53-lys382残基乙酰化表达量呈浓度依存性增加,同时上调p53下游因子p21,mdm2,Bax和PUMA的mRNA水平.另一方面,p53蛋白表达的下调明显削弱了TSA介导的p21表达的上调和cleaved-PARP的产生.结论：组蛋白去乙酰化酶抑制剂TSA能够通过激活p53高乙酰化表达来恢复p53转录功能,从而诱导尤文肉瘤细胞株产生凋亡.     

TSA诱导胰腺癌BxPC-3细胞周期阻滞与凋亡的研究

目的观察曲古抑菌素A(tricho-statin A,TSA)对胰腺癌BxPC-3细胞增殖及凋亡的诱导作用,探讨TSA抗胰腺癌的作用机制。方法BxPC-3细胞应用TSA处理后采用流式细胞技术分析其细胞周期分布和细胞凋亡率,免疫组化方法检测细胞乙酰化组蛋白H4表达,Western blot分析p21 WAF1/CIP1蛋白表达。结果TSA对胰腺癌BxPC-3细胞具有生长抑制作用,且呈时间和剂量依赖性。TSA可将BxPC-3细胞阻滞于G0/G1期,TSA组G0/G1期细胞达(61.8±2.5)%,较空白对照组(42.5±2.2)%和乙醇对照组(47.3±3.4)%明显增多(P=0.004);三组细胞凋亡率分别为25.5%、5.5%和5.1%(P=0.000)。TSA组细胞p21 WAF1/CIP1蛋白表达及其相关染色质乙酰化组蛋白H4表达上调。结论TSA对胰腺癌BxPC-3细胞增殖和细胞周期具有影响作用并可诱导细胞凋亡,p21 WAF1/CIP1蛋白及其相关染色质乙酰化组蛋白H4表达上调可能是其抗胰腺癌作用机制之一。     

NS-398调节HepG2细胞组蛋白乙酰化和p21WAF1/CIP1表达

背景与目的:研究非甾体药物NS-398对人肝癌细胞株HepG2细胞组蛋白H3乙酰化水平的调节作用及对细胞周期素依赖性激酶抑制p21WAF1/CIP1表达的影响.材料与方法:用不同浓度(100、200、300、400μmol/L)的NS-398处理HepG2细胞,以四甲基偶氮唑蓝(MTT)法测定肿瘤细胞增殖抑制率,流式细胞仪(FCM)检测细胞周期的改变及凋亡百分率的变化,应用NS-398分别作用HepG2细胞4、8、12、24、48 h,非药物作用组作为对照,提取细胞的总RNA和总蛋白,采用RT-PCR技术检测p21WAF1/CIP1 mRNA表达情况,并用免疫印迹技术(Western blot)观察组蛋白H3的乙酰化水平变化及p21WAF1/CIP1蛋白的表达水平.结果:NS-398抑制HepG2细胞增殖,且呈剂量依赖性,并诱导其凋亡.且呈浓度依赖性改变细胞周期的分布,一方面增高G0/G1期细胞比例,另一方面降低S期和G2/M期细胞比例,与对照组相比差异具有统计学意义(P＜0.05).NS-398对组蛋白H3乙酰化作用随时间改变而变化,可引起组蛋白H3的乙酰化.NS-398对p21WAF1/CIP1 mRNA和p21WAF1/CIP1蛋白表达的影响呈时间依赖性.结论:NS-398明显上调HepG2细胞组蛋白H3的乙酰化水平,促进细胞周期依赖性激酶抑制剂p21WAF1/CIP1的表达.     

曲古霉素A对肺腺癌细胞株NCI-H1299增殖的抑制作用及其机制

背景与目的：曲古霉素A（trichostatin A,TSA）是具有组蛋白去乙酰化酶（histone deacetylases,HDAC@强效非竞争性抑制剂,对血液系统肿瘤和实质性肿瘤均有较强的生长抑制作用。本文观察HDACs抑制剂TSA对体外培养的肺腺癌NCI-H1299细胞株的增殖、凋亡和周期以及相关基因表达的影响,并探讨其可能的作用机制。方法：MTT法检测不同浓度（0.1、0.2、0.4、2.0μmol/L@的TSA对人肺腺癌NCI-H1299细胞株体外增殖的影响,流式细胞术检测药物处理后细胞周期及凋亡率的变化;Western blot法检测细胞内组蛋白H4乙酰化水平的变化;Real-time PCR检测NCI-H1299细胞内p21、CyclinB1、Bcl-2和Bax的基因表达。结果：TSA能明显抑制NCI-H1299细胞的体外生长,其抑制作用呈明显的剂量和时间依赖性。TSA诱导后,流式细胞术检测结果显示细胞阻滞于G2/M期,细胞凋亡增加。TSA可明显提高NCI-H1299细胞内组蛋白H4的乙酰化水平,诱导p21和Bax的mRNA表达增加,同时抑制Bcl-2和CyclinB1表达。结论：TSA可通过诱导细胞凋亡及阻滞细胞周期而发挥体外抗肺腺癌细胞生长的作用,其机制可能与组蛋白乙酰化水平的提高以及调控相关基因p21、Bax、Bcl-2和CyclinB1的表达变化有关。     

下调Skp2表达诱导乳腺癌MCF-7细胞凋亡

目的 探讨在乳腺癌细胞系MCF-7中下调S期激酶相关蛋白(Skp2)表达诱导细胞凋亡及其机制。方法 应用RNAi方法在体外下调乳腺癌细胞系MCF-7中Skp2的表达水平,48小时后表阿霉素处理细胞,TUNEL和Hoechst 33258染色检测凋亡,Western Blot检测细胞周期调控相关因子及凋亡相关蛋白表达情况,研究其机制。结果 下调MCF-7中Skp2表达水平后,乳腺癌细胞凋亡增加。下调Skp2使p27、p21和CyclinE蛋白表达水平升高。表阿霉素处理MCF-7细胞后,Skp2蛋白水平下调。Skp2 siRNA与表阿霉素有协同诱导凋亡的作用,p53蛋白水平升高。结论 p27、p21和CyclinE在通过下调Skp2诱导的凋亡中发挥作用。Skp2 siRNA和表阿霉素协同诱导细胞凋亡,与p53依赖的凋亡途径有关。Skp2可能是乳腺癌治疗的靶点。     

去甲二氢愈创木酸抑制宫颈癌SiHa细胞的生长与上调p21基因组蛋白乙酰化相关性的研究

目的:研究去甲二氢愈创木酸(nordihydroguaiaretic acid, NDGA)对宫颈癌SiHa细胞增殖的影响及作用机制.方法:MTT法检测NDGA对SiHa细胞生长的影响,FCM法检测NDGA对SiHa细胞周期及凋亡的影响,RT-PCR检测NDGA对p21基因转录的影响,Western印迹法检测NDGA对组蛋白H3总乙酰化水平的影响,染色质免疫沉淀(chromatin immunoprecipitation,ChIP)-PCR法检测NDGA对p21基因近启动子区域组蛋白H3乙酰化的影响.结果:NDGA能明显抑制SiHa细胞的生长,且呈时间和剂量依赖性;可使SiHa细胞的细胞周期阻滞于G1期,而凋亡率却下降;能明显提高p21基因的转录水平和组蛋白H3的总乙酰化水平,ChIP-PCR检测结果表明NDGA可明显促进p21基因近启动子区域组蛋白H3的乙酰化水平.结论:NDGA促进p21基因近启动子区域及细胞内总体组蛋白H3的乙酰化可能是其抑制宫颈癌SiHa细胞生长的潜在机制.     

曲古抑菌素A 对人结肠癌细胞株Colo205 组蛋白乙酰化及ING1b mRNA表达的影响*

目的:探讨曲古抑菌素A（TSA ）对人结肠癌细胞株Colo 205 组蛋白乙酰化及ING 1b mRNA 表达的影响。方法:培养人结肠癌细胞株Colo 205,对照组（A 组）不加TSA 干预,实验组分3 组（B、C、D 组）,分别应用组蛋白去乙酰化酶（HDACs）抑制剂TSA50、100、200 μ g/L 的浓度作用于人结肠癌细胞株Colo 205,24h 后用染色质免疫沉淀（ChIP）方法检测4 组Colo 205 细胞乙酰化组蛋白H3 结合的DNA情况,以了解抑癌基因ING 1b 相关组蛋白H3 乙酰化的变化,并用逆转录聚合酶链反应（RT-PCR）方法检测ING 1b mRNA 的表达,均用实时定量PCR 方法分析。结果:A 组人结肠癌细胞株Colo 205 组蛋白H3 乙酰化水平及ING 1b mRNA Ct值为23.25± 0.08和23.32± 0.05,经TSA 干预后,C、D 组组蛋白H3 乙酰化水平较A 组增加（P<0.05）,2-Δ Δ Ct值分别为4.21和4.38,ING 1b mRNA 表达亦比A 组高（P<0.05）,2-Δ Δ Ct值分别为4.52和4.62,组蛋白H3 乙酰化水平及ING 1b mRNA 表达C、D 组间无显著性差异（P>0.05）,组蛋白H3 乙酰化水平及ING 1b mRNA 表达B 组与A 组比较无显著性差异（P>0.05）,2-Δ Δ Ct值分别为1.12和1.33。同时观察到C、D 组Colo 205 细胞较A、B 组细胞生长明显受抑制。结论:人结肠癌细胞株Colo 205 组蛋白去乙酰化可能是导致基因ING 1b 表达沉默的主要原因之一,100 μ g/L 的TSA 能较好地提高组蛋白乙酰化水平,并有效地激活去乙酰化所致ING 1b 基因转录,诱导该基因表达,从而抑制肿瘤细胞生长。      

曲古抑菌素A和紫杉醇对子宫内膜癌细胞增殖和凋亡的影响

目的: 研究曲古抑菌素A（trichostatin A,TSA）和紫杉醇（paclitaxel, PTX）对人子宫内膜癌细胞株KLE增殖和凋亡的影响。方法：TSA和PTX、卡铂（carboplatin，Carbo） 、多柔比星（doxorubicin，Dox）单独或联合作用于KLE细胞，锥虫蓝法观察药物对肿瘤细胞生长的影响；Annexin V、Hoechst染色和线粒体膜电位检测细胞凋亡；Western blotting检测肿瘤细胞凋亡信号通路中多聚ADP核糖聚合酶（PARP）、半胱天冬氨酸蛋白酶9（caspase9）和乙酰化微管蛋白的表达。结果：PTX、Carbo、Dox和TSA对KLE细胞增殖均有抑制作用，TSA和PTX联用后抑制作用最强。Annexin V染色、Hoechst染色、线粒体膜电位法和PARP、Caspase9的检测显示，单用PTX或TSA均可诱导细胞凋亡, 联合应用产生最强的协同作用。Western blotting和免疫组化分析显示，PTX和TSA均可诱导微管蛋白乙酰化，联合用药后微管蛋白乙酰化明显增加。结论: TSA和PTX联合使用有明显的协同作用，能显著抑制子宫内膜癌KLE细胞生长和诱导细胞凋亡，其机制与激活线粒体凋亡信号通路和增强微管蛋白乙酰化有关。     

非甾体消炎药诱导胃癌细胞凋亡及其机制的研究

目的：明确非甾体消炎药(NSAIDs)能否诱导胃癌细胞凋亡；明确不同的p53基因表型对NSAIDs诱导的细胞凋亡是否有影响；明确NSAIDs对细胞凋亡相关基因Bcl-2及Bax表达的调控。方法：通过MTT比色法检测NSAIDs对细胞生长活力的影响；应用丫啶橙(AO)染色、Annexin-V／PI双染色、共聚焦显微镜、流式细胞术检测细胞凋亡；应用RT-PCR、Western-blot方法检测bcl-2、bax基因及蛋白水平的改变。结果：NSAIDs药物吲哚美辛(Indo)和阿司匹林(Asp)对胃癌细胞株AGS(p53 ／ )、MKN28(p53-／-)均有显著的生长抑制作用，且呈时间／浓度依赖性增强；在相同作用条件下，AGS细胞的凋亡率明显高于MKN28细胞，处理组MKN28细胞凋亡数量虽有所增多，但与正常对照组相比不具有统计学意义；随着药物作用时间的延长，Bcl-2基因mRNA表达逐渐减弱，Bax基因及蛋白表达逐渐增强，在药物作用6～24小时改变最为明显。结论：一定浓度的NSAIDs作用一定时间后，可诱导胃癌细胞凋亡，这为NSAIDs的抗肿瘤应用增加了理论依据；NSAIDs不能诱导p53基因突变的MKN28胃癌细胞株发生显著的凋亡，p53基因突变可能阻断了NSAIDs的凋亡诱导效应；NSAIDs可能通过调控Bcl-2、Bax的基因及蛋白水平而诱导肿瘤细胞凋亡。     

人乳腺癌中p21WAF1组蛋白H3、H4乙酰化水平的表达变化

[目的]探讨人乳腺癌中p21WAF1组蛋白H3、H4乙酰化水平的变化及其意义。[方法]应用HE染色鉴定乳腺癌的病理形态变化,RT—PCR检测p21WAF1mRNA的表达,染色质免疫沉淀法检测p21WAF1组蛋白H3、H4乙酰化的状态。[结果]HE染色可见,与癌旁组织及正常乳腺组织相比,乳腺癌组织结构及细胞形态有明显的异型性。RT—PCR检测结果显示,乳腺癌组织中p21WAF1mRNA的表达水平明显低于癌旁组织及正常乳腺组织（P〈0．05）;p21WAF1mRNA在组蛋白H3、H4乙酰化水平降低乳腺癌中的表达明显低于组蛋白H3、H4乙酰化水平非降低乳腺癌中的表达（P〈0．05）;p21WAF1mRNA表达降低与人乳腺癌的临床分期、分化程度和淋巴结转移有关。染色质免疫沉淀法显示,乳腺癌组织中p21WAF1组蛋白H3、H4的乙酰化水平明显低于癌旁组织及正常乳腺组织;p21WAF1乙酰化水平降低与人乳腺癌的分化程度和淋巴结转移有关。[结论]p21WAF1组蛋白H3、H4乙酰化的表达变化与乳腺癌的发生发展密切相关。     

Association of JNK1 with p21waf1 and p53: modulation of JNK1 activity

Exposure of mammalian cells to genotoxic stress results in activation of the c-jun amino-terminal kinase (JNK)-stress-activated protein kinase (SAPK) pathway and induction of DNA repair enzymes and cell cycle-regulatory proteins such as p53 and p21waf1. The p53 tumor suppressor protein transmits signals that activate p21waf1 gene expression. The p21waf1 protein then restricts cell-cycle progression, thereby allowing time for DNA repair to occur. In this study, we investigated the effects of modulation of the level of wild-type and mutant p53 protein on basal JNK1 activity in the A1-5 rat fibroblast cell line. This cell line contains a p53 gene coding for a temperature-sensitive p53 protein, which allows us to regulate the relative level of wild-type and mutant p53 protein produced in cells. Using the immune complex kinase assay to measure JNK1 activity, we demonstrated that cells expressing the wild-type-conformation p53 protein (when grown at 32.5 degrees C) exhibited a very low level of JNK1 activity. When cells were grown at 37 degrees C or 39 degrees C to express predominantly mutant p53 protein, basal level of JNK1 activity was significantly higher than at 32.5 degrees C. We also demonstrated protein-protein interactions between the p53, p21waf1, and JNK1 proteins in this cell line. Both wild-type p53 protein (expressed at 32.5 degrees C) and mutant p53(val135) protein (expressed at 37 degrees C and 39 degrees C) were present in immunocomplexes of JNK1 protein. Under conditions where wild-type p53 protein was present to induce p21waf1 expression (at 32.5 degrees C), a higher level of p21waf1 protein was also detected in the JNK1 immunocomplexes than in those at 37 degrees C and 39 degrees C. We next investigated the effect that co-association of p53 protein and p21waf1 protein would have on JNK1 activity. We measured basal levels of JNK1 activity in cells expressing wild-type p53 and p21waf1, or in p21waf1-null cells, and demonstrated that cells expressing both p53 and p21waf1 proteins exhibited an approximately threefold lower basal level of JNK1 activity when compared with p21waf1-null cells. To confirm that p21waf1 protein expression in cells resulted in reduced JNK1 activity, we transfected p21waf1-/- cells with a p21waf1 expression vector. We observed that JNK1 activity was inhibited after exogenous p21waf1 protein was expressed in these cells. Our results provide evidence for modulation of the JNK1 pathway by p53 and p21waf1 proteins and support the hypothesis that modulation of JNK1 activity occurred through protein-protein interactions between JNK1, p53, and p21waf1 proteins.     

丁酸钠诱导HT 29结肠癌细胞凋亡及其对 p 53靶基因的调控

目的：〖HT5"SS〗分析丁酸钠对HT29结肠癌细胞p53三个主要靶基因（p21waf1,bax和gadd45）的调控,并探讨其作用机制。〖HT5W〗方法：〖HT5"SS〗HT29细胞常规培养在含有和不含有丁酸钠的培养液中。分别用MTT和流式细胞仪(flow cytometry,FCM) 检测细胞增殖和细胞周期分布,通过形态学观察、亚G1峰的检测和AnnexinVFITC 双标记观察细胞凋亡情况;RTPCR和Western blot分别检测丁酸钠对p21waf1,bax和gadd45三种基因mRNA和蛋白表达水平的影响。〖HT5W〗结果：〖HT5”SS〗丁酸钠以剂量和时间依赖的方式抑制HT29细胞增殖和诱导凋亡,并阻滞细胞于G1期。RTPCR 和Western blot结果显示丁酸钠可以促进p21waf1和bax基因mRNA和蛋白的表达,而对gadd45基因的表达无明显影响。〖HT5W〗结论：〖HT5”SS〗2.5 mmol/L以上浓度的丁酸钠可以抑制HT29细胞增殖并诱导凋亡,该作用可能通过上调p21waf1和bax基因表达而实现。     

Anti-sense oligonucleotide of p21(waf1/cip1) prevents interleukin 4-mediated elevation of p27(kip1) in low grade astrocytoma cells

Elevation of the cyclin-dependent kinase (cdk) inhibitor, p27(kip1) is necessary for Interleukin (IL)-4-mediated growth arrest of human low grade astrocytoma (RTLGA) cells and occurs at 24 h of treatment. Pathways involved in IL4 alteration of p27(kip1) are unknown, however. Here we investigated whether other cdk inhibitors contributed to the actions of IL-4 on RTLGA cells. By 12 h of IL-4 treatment, both cdk4 and cdk2 kinase activities against the retinoblastoma protein (pRb) were reduced and nuclear entry of pRb was prohibited. Twelve-hour cdk complexes contained elevated p21(waf1/cip1) but not p27(kip1), p15(ink4B) or p16(ink4A). IL-4 increased p21(waf1/cip1) but not p27(kip1) mRNA levels, and stimulated luciferase activity of a p21(waf1/cip1) promoter-luciferase reporter. In p53-mutant WITG3 cells, IL-4 did not alter p21(waf1/cip1) mRNA and promoter-luciferase activity or p27(kipl) protein, suggesting a need for functional p53. STAT6 phosphorylation by IL-4, however, occurred in both p53-mutant WITG3 and p53-functional RTLGA cells. Pre-treatment of RTLGA with anti-sense but not missense p21(waf1/cip1) oligonucleotide prior to IL-4: (a) restored cdk activities; (b) reduced cdk4-associated p21(waf1/cip1) levels; (c) prevented p27(kipl) elevation; and (d) reversed growth arrest. These results are the first to suggest that p21(waf1/cip1) is essential for IL-4-mediated elevation of p27(kip) and growth arrest of astrocytoma cells.     

Okadaic acid mediates p53 hyperphosphorylation and growth arrest in cells with wild-type p53 but increases aberrant mitoses in cells with non-functional p53.

The protein phosphatase inhibitor and tumor promoting agent okadaic acid (OA), has been shown previously to induce hyperphosphorylation of p53 protein, which in turn correlated with increased transactivation or apoptotic function. However, how the tumor promotion effects of OA relate to p53 tumor supressor function (or dysfunction) remain unclear. Rat embryonic fibroblasts harboring a temperature-sensitive mouse p53 transgene were treated with 50 nM doses of OA. At the wild-type permissive temperature this treatment resulted in: (i) the hyperphosphorylation of sites within tryptic peptides of the transactivation domain of p53; (ii) an increase in p53 affinity for a p21(waf1) promotor oligonucleotide; (iii) an increase in cellular steady state levels of p21(waf1) message; (iv) a G2/M cell cycle blockage in addition to the G1/S arrest previously associated with p53; and (v) no increased incidence of apoptosis. On the other hand, OA treatment at the mutated p53 permissive temperature resulted in a relatively high incidence of aberrant mitosis with no upregulation of p21(waf1) message. These results suggest that while wild-type p53 blocks the proliferative effects of OA through p21(waf1)-mediated growth arrest, cells with non-functional p53 cannot arrest and suffer relatively high levels of OA-mediated aberrant mitoses.     

A p21(waf1)-independent pathway for inhibitory phosphorylation of cyclin-dependent kinase p34(cdc2) and concomitant G(2)/M arrest by the chemopreventive flavonoid apigenin.

Apigenin, a nonmutagenic flavonoid, has been shown to inhibit ultraviolet light-induced skin tumorigenesis when topically applied to mouse skin. Our previous studies have shown that apigenin treatment of cultured mouse keratinocytes induces G(2)/M arrest accompanied by an increase in p53 protein stability and expression of p21(waf1). In this study, we determined whether the G(2)/M arrest induced by apigenin was dependent upon the presence of the cyclin dependent kinase inhibitor p21(waf1). We exposed WWT.8 (p21(waf1) wild-type) and WKO.16 (p21(waf1) null) mouse keratinocytes to various doses of apigenin for 24 h and observed G(2)/M arrest in both cell lines, thereby establishing that the apigenin-induced G(2)/M arrest was p21(waf1) independent. A 4-h treatment with apigenin induced increases in p53 protein level by sixfold and tenfold in the WWT.8 p21(waf1) wild-type cells and WKO.16 p21(waf1) null cells, respectively. After 24 h in WWT.8 cells, p21(waf1) protein also was induced in a dose-dependent manner, but it was not expressed in WKO.16 keratinocytes. We then measured the effect of apigenin treatment on the mammalian homologue of the yeast cdc2 gene (p34(cdc2)) cyclin-dependent kinase and cyclin B1 (cycB1), because these proteins complex to regulate G(2)/M progression. Apigenin treatment decreased the protein level of p34(cdc2), and p34(cdc2) kinase activity was inhibited in both p21(waf1)(+/+) and p21(waf1)(-/-) cell lines by approximately 40%. The inhibition of p34(cdc2) kinase activity by apigenin treatment correlated with increasing levels of p34(cdc2) phosphorylation at Tyr15, a site in the p34(cdc2) kinase that undergoes inhibitory phosphorylation by Wee1 kinase. Apigenin treatment also had no effect on the protein level or activity of the competing phosphatase, cdc25c, which dephosphorylates p34(cdc2) kinase at Tyr15. Apigenin had little effect on the accumulation of cycB1 protein. These results supported the conclusion that G(2)/M arrest induced by apigenin was accompanied by inhibition of the p34(cdc2) cyclin-dependent kinase protein level and activity in a p21(waf1)-independent manner.     

淋巴结阴性乳腺癌p21~(waf1)、c-erbB-2和p53蛋白表达及其临床意义

目的：探讨腋淋巴结阴性乳腺癌组织中ｐ２１ｗａｆ１、ｃ－ｅｒｂＢ－２和ｐ５３基因蛋白表达及其与临床病理参数和预后的关系。方法：应用免疫组化ＬＳＡＢ方法检测１２１例腋淋巴结阴性乳腺癌石蜡切片中ｐ２１ｗａｆ１、ｃ－ｅｒｂＢ－２和ｐ５３蛋白的表达情况;同时应用Ｋａｐｌａｎ－Ｍｅｉｅｒ法及多变量Ｃｏｘ比例风险模型,分析３种蛋白表达与预后的关系。结果：（１）ｐ２１ｗａｆ１蛋白表达率为 ４８． ８％,与病理组织学分级、ＥＲ状态有关; ｐ５３蛋白表达率为 ３６． ４％, ｃ－ｅｒｂＢ－２蛋白表达率为 ２６． ４％,与组织学分级有关;（２）ｐ２１ｗａｆ１阳性表达与 ｐ５３表达呈负相关（ Ｐ＜ ０． ０５）;（３）ｐ２１ｗａｆ１阳性组患者无瘤生存率高于阴性组（Ｐ＜０．０５）;ｃ－ｅｒｂＢ－２阳性组患者无瘤生存率明显低于阴性组（Ｐ＜０ ０１）;Ｃｏｘ模型分析显示仅有ｃ－ｅｒｂＢ－２表达对预后有显著影响。结论：乳腺癌组织ｐ２１ｗａｆ１、ｃ－ｅｒｂＢ－２表达与病理组织学分级有相关性;ｐ２１ｗａｆ１表达依赖于ｐ５３途径刺激; ｐ２１ｗａｆ１、、ｃ－ｅｒｂＢ－２表达与腋淋巴结阴性乳腺癌预后有关,且ｃ－ｅｒｂＢ－２表达是一个独立的预后指标。     

The prognostic significance of p53, p27 kip1, p21 waf1, HER-2/neu, and Ki67 proteins expression in gastric cancer: a clinicopathological and immunohistochemical study of 121 Arab patients

BACKGROUND: The variability of prognosis within a pathological stage of gastric cancer (GC) at presentation, underscores the need for specific biological markers to identify subgroups of patients with aggressive course for intensive treatment. To our knowledge, this is the first study from an Arab population reporting on the relationship of p53, p27 kip1, p21 waf1, HER-2/neu, and Ki67 expression, and clinicopathological features and their prognostic significance. METHODS: Formalin-fixed paraffin-embedded tumors were studied by immunohistochemistry, using monoclonal antibodies to p53, p27 kip1, p21 waf1, HER-2/neu, and Ki67. The results were correlated with clinicopathological features and survival. RESULTS: M:F = 80:41; median age = 60 years; stage III and IV = 71%; and median follow-up = 34.4 months. Positive expression rates of p53, p27 kip1, p21 waf1, Ki67, and HER-2/neu were 54%, 40%, 8.3%, 70%, and 12% respectively. p53 expression correlated with age <60 years (P = 0.03), tumor size >5 cm (P = 0.01), p27 kip1 and Ki67 expression (P = 0.0001), and HER-2/neu (P = 0.04). p21 waf1 correlated inversely with T-stage (P = 0.008) and Her-2/neu expression correlated with histological grade (P = 0.04) and T-stage (P = 0.008). Univariate analysis showed that p53 overexpression (P = 0.01), fungating and infiltrative macroscopic appearance (P = 0.02), size >5 cm (P = 0.0001), lymph node metastasis (P = 0.0001), p T3 and T4 disease (P = 0.01), and overall stage III and IV (P = 0.0001) disease were adverse prognostic factors. Patients with tumor profiles p53 (-)/p27 (+) had better survival than those with p53 (+)/p27 kip1 (-)(P = 0.02). On multivariate analysis by Cox regression model, the expression of p53 (P = 0.03) and lymph node involvement (P = 0.01) were significant adverse prognostic factors for overall survival. CONCLUSIONS: The expression of p53 in Arab patients with GC correlates with aggressive tumor characteristics and is an independent prognostic factor. The combined analysis of p53 and p27 kip1 is of added prognostic value.     

FHIT、p21waf1/cip1基因在膀胱移行细胞癌中的表达与意义

目的:通过测定脆性组氨酸三联体基因(FHIT)及p21waf1/cip1基因在膀胱移行细胞癌组织、正常膀胱组织中的表达,探讨FHIT基因以及p21waf1/cip1基因与膀胱癌的关系及其临床意义。方法:采用免疫组织化学SP法对43例膀胱移行细胞癌(BTCC)组织及14例正常膀胱组织中的FHIT基因及p21waf1/cip1基因的蛋白表达进行检测。结果:FHIT蛋白表达与肿瘤的分期、分级无相关性(P>0.05)而p21waf1/cip1蛋白的表达与之有相关性(P<0.05);FHIT蛋白的表达在GI肿瘤、浅表性肿瘤中明显低于在正常膀胱组织中的表达(P<0.05)而p21waf1/cip1蛋白的表达在上述组织比较中无差别(P>0.05);FHIT蛋白的表达在初发肿瘤中与复发肿瘤中无明显差别(P>0.05)而p21waf1/cip1蛋白的表达在上述两种组织比较有明显差别(P<0.05)。FHIT蛋白与p21waf1/cip1蛋白的表达没有相关性(P>0.05)。结论:FHIT基因可能成为早期诊断膀胱移行细胞癌的指标。p21waf1/cip1基因可能成为估计膀胱移行细胞癌的恶性程度及肿瘤侵袭性、预后的指标。FHIT基因在膀胱移行细胞癌中的作用机制可能与p21waf1/cip1基因没有关系。