Up-regulation of p27Kip1 Correlates Inversely with Anchorage-independent Growth of Human Cancer Cell Lines

We examined the correlation between anchorage-independent growth and cell cycle-related molecules using 39 human cancer cell lines. They consisted of lung-, colon-, stomach-, breast-, ovarian-, brain-, renal- and melanoma-derived cell lines. Their anchorage-independent growth ability varied, but was not clearly related to the tissue of origin. There was a tendency for the levels of cyclin D1, cyclin E, cyclin A, p27, and p21 to show a tissue-dependent expression pattern. Statistical analysis revealed an inverse correlation of the p27 level with anchorage-independent growth ( r =−0.456, P <0.01). Thus, the regulation of p27 is suggested to be linked to the anchorage independence of human cancer cells.     

Expression of Cyclin-dependent Kinase Inhibitor p27/Kipl and AP-1 Coactivator p38/Jabl Correlates with Differentiation of Embryonal Rhabdomyosarcoma

Cyclin-dependent kinase (CDK) inhibitor p27/Kip1 (p27) is a diagnostic and prognostic marker of various malignancies. Low expression of p27 reflects poor differentiation and poor prognosis, and an inverse correlation between the expression of p27 and degree of tumor malignancy has been reported. Because p27 mutation is extremely rare in human tumors, it is important to study the expression of p27 and its inactivator, p38/Jab1 (JAB1). Here we analyzed the expression of p27 and JAB1 by immunohistochemistry in embryonal rhabdomyosarcoma (E-RMS). We first confirmed the expression of p27 and JAB1 in normal human tonsillar epithelium, and observed a coordinated expression pattern depending on cell differentiation. Subsequently, specimens of eight poorlyand three well-differentiated E-RMS were examined for the expression of p27 and JAB1. The analyses revealed that four out of eight poorly-differentiated E-RMS were negative for p27, with positivity for nuclear JAB (NJAB) (- /± for p27/NJAB) in three and negativity for any JAB-1 expression (- / -) in one. The remaining four poorly-differentiated E-RMS expressed p27 in the nuclei, together with predominant NJAB (±/±). In three well-differentiated E-RMS, only one expressed nuclear p27 and all of these three expressed no NJAB (±/ - for p27/NJAB), but expressed predominant cytoplasmic JAB1 (CJAB). These findings suggest that JAB1 may play an important role in determining the differentiation stage of rhabdomyosarcoma cells by modulating the activity of CDK inhibitor p27.     

宫颈癌中p27Kip1基因的表达与体外药物敏感性的关系

背景与目的：p27^Kip1基因是调控细胞周期并抑制细胞分裂的重要基因。它与诱导肿瘤细胞的凋亡和化疗药物的耐药也有关系。本文旨在探讨宫颈癌组织中p27^Kip1基因的表达与体外药敏的关系。方法：采用MTT法检测了部分化疗药物在46例宫颈癌组织中的抑制率，并采用免疫组织化学方法检测了癌组织中p27^Kip1基因的表达。结果：宫颈癌组织中p27^Kip1表达的阳性率为15.2％（7／46）。顺铂（DDP）、表柔比星（E—ADM）、替尼泊苷（VM26）、紫杉醇（Taxol）、吉西他滨（Gemzar）、氟尿嘧啶（5-FU）和BLM的平均抑制率分别为47.6％、38．0％、42．7％、27．4％、23.0％、26.3％和24．8％。DDP、VM26和Taxol在p27^Kip1表达阴性的宫颈癌标本中的抑制率显著低于阳性标本（P〈0.05），而在其他药物中差异无显著性。结论：p27^Kip1的表达可作为判断肿瘤对化疗药物的敏感性的指标，指导临床用药。     

胃癌中Cks1 、p27 Kip1和Skp2 蛋白的表达

 目的 探讨Cks1在胃癌发生及其在Skp2调节p27 ^Kip1降解过程中的作用。方法 应用流式细胞术测定正常胃粘膜和胃癌组织中Cks1、p27^Kip1、Skp2蛋白表达。结果胃癌组织中Cks1、Skp2表达明显高于正常胃粘膜（x²=22．69，P〈0．05；x²=13．42，P〈0．05），而p27 ^Kip1表达则低于正常胃粘膜（x²=14．83，P〈0．05）。胃癌中Cks1、Skp2表达与p27 ^Kip1表达呈负相关（r=-0．649，P〈0．05；r=-0．732，P〈0．05）；而Cks1蛋白表达与Skp2蛋白缺乏相关性（P〉0．05）。胃癌中Cks1的表达与肿瘤分化程度相关（x²=5．05，P〈0．05），而与肿瘤浸润深度、淋巴结转移及临床分期不相关（P〉0．05）。结论Cks1可能参与了胃癌的发生；胃癌中Cks1可能参与了Skp2调节p27 ^Kip1泛素化降解过程。     

口腔鳞状细胞癌细胞周期因子p27 cyclin D1及CDK4的表达与意义

目的：从细胞周期调控的角度来探讨口腔鳞状细胞癌（OSCC）的发生、发展及预后和p27、cyclin D1和CDK4的关系。方法：采用免疫组织化学方法，检测了50例口腔鳞状细胞癌及10例正常口腔粘膜中p27、cyclinD1和CDK4蛋白表达的水平，然后用Spearman 相关分析检查它们与临床病理学指标的关系以及它们三者之间的相关关系，用Kaplan-Meier方法绘制生存曲线并进行生存分析，Cox比例风险模型作预后分析。结果：1)p27在所有正常口腔粘膜上皮均呈现高表达，而在口腔鳞状细胞癌组织表达降低，p27的低表达与临床分期，淋巴结转移有显著性相关关系，cyclin D1和CDK4在正常粘膜上皮呈现低水平表达，在口腔钙状细胞癌组织中过表达。2)cyclinD1和CDK4在正常粘膜上皮呈现低水平表达，在口腔鳞状细胞癌组织中过表达.2)cyclin D1的表达与CDK4呈正相关（r=0.442,P=0.001）;p27与CDK4的表达呈负相关（r=-0.384,P=0.006）.3)Kaplan-Meier生存分析显示p27高表达组（ ）的各期的生存均高于低表达组（-）,cyclin D1染色（ ）组的生存率低于（-）组。4）Cox比例风险模型分析结果显示p27蛋白表达水平，cyclinD1蛋白表达水平，淋巴结转移和临床分期分别是口腔鳞状细胞癌的独立预后指标。结论：口腔鳞状细胞癌组织中，p27,cyclin D1和CDK4蛋白的异常表达说明它们均参与了口腔鳞状细胞癌的发生发展，且在这一过程中三者之间存在相互协同与制约关系。在临床应用中有可能将p27和cyclin D1蛋白的表达程度作为判断口腔鳞状癌患者预后的指标。     

褪黑素对小鼠肝癌细胞增殖及p27Kip1、cyclin D1基因表达的影响

目的探讨褪黑素(melatonin MT)抑制H22小鼠肝癌细胞增殖的可能机制.方法通过体外细胞培养,分别采用不同剂量MT作用不同的时间,MTT显色技术检测细胞的增殖;RT-PCR检测MT作用4 d后,H22细胞中p27Kip1、cyclin D1mRNA的表达;免疫组化检测H22细胞中p27Kip1、cyclin D1蛋白的表达.结果MT具有抑制肝癌细胞的生长和增殖的作用,且具有时间依赖关系.MT诱导细胞凋亡的过程中,与对照组相比,MT药理剂量组H22细胞中的p27Kip1 mRNA及蛋白表达升高(P＜0.01);cyclin D1 mRNA及蛋白表达下降(P＜0.01).MT生理剂量组p27Kip1mRNA表达升高(P＜0.01),而p27Kip1蛋白表达水平与对照组相比差异无显著性(P＞0.05),cyclinD1 mRNA及蛋白表达均下降,差异元显著性(P＞0.05).结论MT抑制肝癌细胞的增殖可能与上调细胞周期抑制因子p27Kip1的表达,降低周期蛋白cyclin D1表达,进而延迟细胞周期的进程有关.     

Troglitazone Induces G1 Arrest by p27Kip1 Induction That Is Mediated by Inhibition of Proteasome in Human Gastric Cancer Cells

We examined in the present study whether human gastric cancer cells express peroxisome proliferator-activated receptor γ (PPARγ), the effect of PPARy activation by troglitazone, a selective ligand, on cellular growth, and the mechanism of the growth arrest by troglitazone in gastric cancer cells. RT-PCR, northern blot and western blot analysis demonstrated that all four tested human gastric cancer cell lines, MKN–28, MKN–45, MKN–74 and KATO-III, expressed PPARγ mRNA and protein. WST–1 assay and flow cytometric analysis revealed that troglitazone inhibited the growth and induced G1 arrest in all four gastric cancer cell lines. To examine the role of p27^Kip1, a cyclin-dependent kinase inhibitor, in the G1 arrest by troglitazone, we determined p27^Kip1 protein expression by western blot analysis in gastric cancer cells that had been treated with troglitazone. Troglitazone increased p27^Kip1 in all four gastric cancer cell lines. Since it has been reported that the ubiquitin-proteasome system plays a vital role in the degradation of p27^Kip1 protein, we evaluated the hypothesis that inhibition of proteasome mediates the troglitazone-induced p27^Kip1 accumulation. Lactacystin, a proteasome inhibitor, inhibited cell growth and increased p27^Kilp1 expression in MKN–74 cells. It was further demonstrated that troglitazone inhibited proteasome activity in a dose-dependent manner in MKN–74 cells. All these results suggest that troglitazone inhibited proteasome activity, followed by induction of p27^Kipl, which arrests cells at the Gl phase of the cell cycle in gastric cancer cells. The troglitazone-mediated inhibition of the proteasome suggests a novel mechanism for the anti-proliferative effect of this agent in cancer cells.     

乳腺癌组织p27Kip1的表达及其与细胞增殖的关系

目的:研究乳腺癌组织中细胞周期抑制剂p27Kip1的表达及其意义,并探讨其与细胞增殖的关系.方法:应用免疫组化SP法检测80例乳腺癌和20例癌旁正常组织中p27Kip1和增殖细胞核抗原(pro-liferative cell nuclear antigen,PCNA)的表达.结果:乳腺癌组织中p27Kip1高表达率为53.75%(43/80),明显低于癌旁正常组织(P<0.01).p27Kip1表达与组织学分级、TNM分期及淋巴结转移均相关,P<0.05.乳腺癌组织中p27Kip1的表达与PCNA LI呈负相关,r=-0.372,P<0.05.p27Kip1高表达的乳腺癌术后5年无病生存率(DFS)明显高于p27Kip1低表达者(P<0.05).结论:p27Kip1表达缺失可促进肿瘤细胞增殖,是判断乳腺癌发生、发展及预后的有效生物学指标.     

胃癌中p27Kip1、p53表达与其浸润、转移和预后的关系

目的研究胃癌组织中p27Kip1和p53的表达与胃癌浸润、转移和预后的关系。方法用免疫组化(二步法)检测100例胃癌组织中的p27Kip1蛋白和p53蛋白的表达。结果100例胃癌组织中p27Kip1和p53蛋白阳性表达率分别为44%和49%。在胃癌深部浸润组、淋巴结转移组和5年内死亡组中p27Kip1呈显著低表达(P<0.05);在胃癌淋巴结转移组和5年内死亡组中p53呈显著高表达(P<0.05)。经单变量分析结果显示p27Kip1高表达组5年生存率为70.59%,显著高于低表达组54.55%,和阴性组26%;p53高表达组5年生存率为19.23%,显著低于低表达组43.75%和阴性组53.19%。多变量Cox回归模型分析显示p27Kip1、p53均是独立的预后指标,但p27Kip1表达对患者预后的相对危险度(RR=3.06)显著大于p53表达的相对危险度(RR=2.33,P<0.01)。结论p27Kip1低表达与p53高表达的癌细胞常更能浸润与转移,使患者生存率明显降低。p27Kip1和p53是胃癌预后的独立指标,其中p27Kip1表达评估胃癌预后的作用优于p53。     

P27蛋白在人脑胶质瘤组织中的表达及其临床意义

Objective： To study p27/Kipl expression in gliomas and its application value. Methods： Imo munohistochemical technique was used to detect the exprssion of p27/Kipl gene in 48 different malignant grade human brain glioma tissues categorized according to WHO classification and 12 normal human brain tissues,which were analyzed quantitatively by using the image system and compared retrospectively with the patients＇ clinical characteristics. Results： In this series, the immunohistochemical reaction for p27/Kipl was confined to the nuclei. The abnormal positive expression rate of p27/Kipl in gliomas was found to be higher than that in normal tissues （P〈0.05）. The positive nuclei expression of p27/Kipl decreased in number and staining intensity with the increasing degree of histological malignancy （P〈0.05）. Lower expression of p27/Kipl was associated with poor prognosis （P〈0.05）. P27/Kipl expression could be regarded as an independent prognostic factor. Conclusion： The abnormal expression of p27/Kipl may be closely related to the occurrence and development of gliomas, and also can be used to evaluate the prognosis independently.     

Cell cycle regulators p27 and pRb in lymphomas - correlation with histology and proliferative activity

The cell cycle is a complex event in which multiple regulator-proteins participate. The G(1)/S checkpoint of the cell cycle is controlled by pRb protein, which functions in its hypophosphorylated form as a negative regulator of growth. p27 (Kip1), a member of CIP/KIP family of cyclin inhibitory proteins, participates in inhibition of forming complexes that allow pRb to phosphorylate and lead the cell into mitosis. The expression of these important cell cycle regulator proteins was studied in a total of 96 non-Hodgkin's lymphoma (NHL) samples, which were classified according to the REAL classification. The expression of p27, pRb and the cell proliferation marker Ki-67 (MIB-1) was evaluated in lymphomas using immunohistochemistry. This study showed that there were coordinate changes in the expression of p27 and pRb in NHL. When compared to low-grade lymphomas, high-grade lymphomas showed significantly reduced expression of p27 and inversely pRb expression was increased (P< 0.001). Increase in expression of Ki-67 was parallel with pRb expression, and was mainly seen in cells that lacked p27 expression (P< 0.0001). This study suggests that changes in the control of the cell cycle closely relate to the pathobiology of NHL.     

Expression of p21waf-1/cip-1 Is Significantly Induced in the Livers of LEC Rats with Chronic Liver Injury

It is reported that hepatocytes isolated from LEC rats with chronic liver injury show reduced growth activity in primary culture. To elucidate the molecular basis of this phenomenon, we examined expression of p21^waf-1/cip-1 and p27, cyclin-dependent kinase inhibitors, by northern blot analysis. The expression of p21^waf-1/cip-1 in the LEC rat liver was 3-fold higher than that of age-matched SD rat liver, while there was no significant difference in p27 expression level. Western blot analysis also revealed a significant increase in p2l^waf-1/cip-1 in the nuclear matrix fraction of the LEC rat liver. Immunohisto-chemically, p21^waf-1/cip-1 was detected in the nuclei of normal LEC rat hepatocytes, but not in those of hepatocellular carcinoma cells, suggesting selective growth of neoplastic hepatocytes.     

p27~(Kip1)在宫颈癌中的表达及其临床意义

[目的]研究子宫颈癌手术标本中p27Kip1 的表达在肿瘤预后判断中的临床意义。[方法]应用免疫组化SP法 ,检测宫颈癌手术标本中p27Kip1 的表达 ;[结果]p27Kip1 的表达与癌组织病理分期呈负相关 ,与肿瘤的预后呈正相关。[结论]子宫颈癌组织中 ,p27Kip1表达缺失的肿瘤组织恶性程度更高 ,并且直接影响肿瘤患者的术后生存期 ,是影响肿瘤预后的有效参考指标。检测癌组织中p27Kip1 的表达情况 ,有助于估计患者的预后 ,指导患者术后的综合治疗。     

乳腺癌中细胞周期依赖性蛋白激酶抑制蛋白p27Kip1表达水平

目的探讨p27Kip1蛋白在乳腺癌中的表达水平与乳腺癌生物学行为的关系和对预后的影响.方法用免疫组化法检测了132例乳腺癌和16例正常乳腺组织中p27Kipl蛋白的表达水平,对其中有完整随访资料的90例乳腺癌进行了临床病理学特征和生存时间关系的分析.结果p27Kip1蛋白在正常乳腺组织中高表达率为100%(16/16);在乳腺癌中高表达率为40.15%(53/132),低表达率为59.85%(79/132),差异有显著性(P＜0.005).不同类型的乳腺癌中p27Kip1蛋白的表达水平差异无显著性(P＞0.1).乳腺癌组织p27Kipl蛋白表达水平与病理组织学分级、肿瘤内坏死、核分裂数、腋淋巴结转移情况、局部复发和远处转移显著相关(P＜0.005),与肿块大小、年龄、ER水平、月经状况无显著关系.生存关系分析显示p27Kip1蛋白低表达的乳腺癌患者生存率明显低于p27Kipl蛋白高表达者(P＜0.001).Cox模型则显示p27Kip1蛋白是乳腺癌的一个独立的预后标志物.结论①p27Kip1蛋白的表达水平与乳腺癌的组织学类型无关;②p27Kip1蛋白是检测乳腺癌预后有价值的独立指标,有助于临床选择高危复发病例;③p27Kip 1蛋白表达水平可做为术后进一步治疗的参考指标.     

p27kip1与细胞周期素D1在子宫内膜癌中的表达

[目的]研究p27kip1和细胞周期素D1蛋白在子宫内膜癌中的表达及其意义。[方法]采用免疫组化LSAB法检测42例子宫内膜癌中p27kip1、细胞周期素D1的表达。[结果]42例子宫内膜癌中20例p27kip1表达阳性,占47.6%,p27kip1与子宫内膜癌的细胞分级、临床分期、肌层浸润深度有关(P<0.05);17例子宫内膜癌细胞周期素D1表达阳性,占40.4%,细胞周期素D1与子宫内膜癌的细胞分级、临床分期、淋巴结转移有关(P<0.05);p27kip1、细胞周期素D1协同表达15例,均为晚期或低分化癌。[结论]p27kip1、细胞周期素D1作为细胞周期调节因子参与子宫内膜癌的发生、发展,其协同作用促进子宫内膜癌的发展,且预后不良。     

Skp2、p27Kip1在胆囊癌组织中的表达及其临床意义

目的探讨Skp2、p27Kip1在胆囊癌及癌旁组织中的表达情况及与其临床病理特征的关系。方法采用免疫组织化学PV6000二步法,检测Skp2、p27Kip1在78例胆囊癌及癌旁组织中的表达情况,并应用SPSS13.0统计软件分析2种生物指标与胆囊癌临床参数的关系。结果 Skp2与p27Kip1在胆囊癌中的阳性表达率分别为59.0%和39.7%,在癌旁组织中为25.0%和70.0%,胆囊癌中Skp2、p27Kip1阳性表达率均高于胆囊癌旁组织(P均<0.05)。Skp2表达与Nevin分期、分化程度及坏死情况相关,Nevin分期高、分化程度低及伴有坏死的患者Skp2表达高(P均<0.05)。p27Kip1阳性表达与患者Nevin分期、神经侵犯及坏死情况呈负相关性,Nevin分期早、无神经侵犯或不伴坏死患者p27Kip1表达水平高(P均<0.05)。相关性检验显示Skp2与p27Kip1两者间无相关性(P>0.05)。结论 Skp2、p27Kip1均与胆囊癌的发生发展有一定关系,可能成为胆囊癌新的临床诊断和监测预后的指标.     

胃癌中p27~(Kip1)、p53表达与其浸润、转移和预后的关系(英文)

目的研究胃癌组织中p27~(Kipl)和 p53的表达与胃癌浸润、转移和预后的关系。方法用免疫组化(二步法)检测100例胃癌组织中的 p27~(Kipl)蛋白和 p53蛋白的表达。结果 100例胃癌组织中 p27~(Kipl)和 p53蛋白阳性表达率分别为44%和49%。在胃癌深部浸润组、淋巴结转移组和5年内死亡组中p27~(kipl)呈显著低表达(P<0.05);在胃癌淋巴结转移组和5年内死亡组中 p53呈显著高表达(P<0.05)。经单变量分析结果显示p27~(Kipl)高表达组5年生存率为70.59%,显著高于低表达组54.55%和阴性组26%;p53高表达组5年生存率为19.23%,显著低于低表达组43.75%和阴性组53.19%。多变量 Cox 回归模型分析显示p27~(Kipl)、p53均是独立的预后指标,但 p27~(Kipt)表达对患者预后的相对危险度(RR=3.06)显著大于 p53表达的相对危险度(RR=2.33,P<0.01)。结论 p27~(Kipl)低表达与 p53高表达的癌细胞常更能浸润与转移,使患者生存率明显降低。p27~(kipl)和 p53是胃癌预后的独立指标,其中 p27~(Kipl)表达评估胃癌预后的作用优于 p53。     

胶质瘤p27Kip1、bcl-2和PCNA蛋白的表达

目的　探讨胶质瘤p2 7Kip1,bcl 2和PCNA蛋白表达与肿瘤恶性程度、细胞增殖活性、凋亡程度的关系。方法　采用免疫组化染色S P法检测 66例不同级别的胶质瘤 p2 7Kip1,bcl 2和PCNA蛋白的表达。结果　在 66例胶质瘤中 ,p2 7Kip1表达 18例(2 7% ) ,bcl 2表达 2 0例 (3 0 % ) ,PCNA表达 5 1例 (77% )。p2 7Kip1蛋白表达率随着胶质瘤级别升高而减少 ;bcl 2蛋白表达率随肿瘤级别升高而相应增加 ;PCNA表达随胶质瘤级别升高阳性反应强度增加 ;但Ⅰ、Ⅱ与Ⅲ级和Ⅳ级组间无显著性差异。结论　p2 7Kip1蛋白表达的缺失可能与胶质瘤的发生有关 ;bcl 2基因可能间接抑制细胞凋亡而与胶质瘤的分型、细胞的增殖活性以及潜在的临床行为无直接关系 ;PCNA的表达与星形胶质细胞瘤的恶性行为有关     

p53 Gene Mutation in the Bone-Marrow of a Patient with Diffuse Mixed Cell Type Lymphoma at Diagnosis Predicting Eventual Progression to Large Cell Lymphoma

Mutations of the p53 tumor suppressor gene have been used as molecular genetic markers of disease and serve as a prognostic indicator in various malignancies including non-Hodgkin's lymphoma (NHL). Alterations in the p53 gene were investigated in a bone marrow sample from a NHL patient admitted for autologous bone marrow transplantation. Diffuse mixed small and large cell NHL, was initially diagnosed which eventually progressed to large cell lymphoma at relapse following poly-chemotherapy. A sequential technique of polymerase chain reaction-mediated single-strand conformational polymorphism (PCR-SSCP) of the p53 gene revealed a shift in one band of exon 6 in the bone marrow, collected at the time of initial diagnosis. No mutations were detected in exons 5, 7, 8 and 9. Direct sequencing of exon 6 detected a single base change from G to C resulting in an amino acid substitution from glycine to histidine. Results of this study and data reviewed from other publications suggest that the missense p53 mutation seen in this patient at the time of diagnosis may perhaps have been used to predict the eventual outcome of the disease. This could, therefore, serve as an important genetic disease marker particularly in bone marrow or peripheral blood samples initially collected and cryopreserved for future possible autologous transplantation.