TAM诱导MA782细胞凋亡与Cyclin D1、CDK4、TGF-β1的关系

目的 :探讨Tamoxifen(TAM )诱导ER(- )小鼠乳腺癌细胞MA782细胞凋亡及其分子作用机制。方法 :TAM体外作用于MA782细胞 ,用MTT法检测细胞增殖活性、流式细胞仪检测细胞凋亡和细胞周期分布 ,免疫组化检测cyclinD1、CDK4、TGF β1蛋白表达 ,并用病理图像分析软件进行半定量分析。 结果 :TAM在体外能明显抑制MA782细胞生长 ,诱导细胞凋亡 ,下调cyclinD1、CDK4表达 ,上调TGF β1表达。免疫组化半定量分析显示 6μmol/L作用 72小时后 ,cyclinD1、CDK4蛋白平均灰度值分别从 132 .5± 0 .0 2、10 7.2± 0 .0 1下降至 12 6 .18± 0 .0 3、76 .2 1± 0 .0 3,10 μmol/L作用 72小时后分别为 73.5 6± 0 .0 2、72 .0 3± 0 .0 1,与对照组相比差异极其显著性 (P <0 0 1)。MA782细胞TGF β1平均灰度值为 5 9.72± 0 .0 2 ,2 μmol/L作用 4 8小时后无明显变化 ,72小时后上升到97 1± 0 .0 3,6、10 μmol/L作用 4 8小时后平均灰度值分别为 83.2± 0 .0 4、96 .83± 0 .0 2 ,72小时后分别上升到12 1 75± 0 .0 3、139.0 1± 0 .0 5 ,与对照组相比差异极其显著 (P <0 .0 1)。结论 :TAM诱导MA782细胞凋亡其分子作用机制可能与下调cyclinD1、CDK4蛋白表达 ,上调TGF β1蛋白表达有关。     

颌骨骨肉瘤中cyclinD1和CDK4的表达及其意义

分析细胞周期素Ｄ１（ｃｙｃｌｉｎＤ１）和细胞周期素依赖激酶４（ＣＤＫ４）在颌骨骨肉瘤中的表达及意义。方法采用免疫组化ＡＢＣ法检测ｃｙｃｌｉｎＤ１和ＣＤＫ４在２０例颌骨骨肉瘤和８例骨软骨瘤中的表达。结果骨肉瘤中ｃｙｃｌｉｎＤ１和ＣＤＫ４的阳性率表达率分别为６５．００％（１３／２０）和６０．００％（１２／２０），两者的阳性表达存在正相关（γＳ＝０．４８，Ｐ〈０．０５）；而它们在骨软骨瘤中的阳性率均为１     

细胞周期蛋白cyclin D1、CDK4在食管癌中的表达及其意义

目的：获得食管癌发生过程中调节Ｇ１细胞周期各种因子的作用。方法：采用抗ｃｙｃｌｉｎＤ１和ＣＤＫ４的单克隆抗体对１０例下沉食管和５０例食管鳞状上皮癌标本进行免疫组织化学染色。结果：ｃｙｃｌｉｎＤ１和ＣＤＫ４在正常食管上皮呈现较低水平的表达,在食管鳞状上皮癌中则过表达。２７／５０食管鳞状上皮癌ｙｃｙｃｌｉｎＤ１染色阳性,其中８例强阳性,１２例只表达ＣＤＫ４,１１例只表达ｃｙｃｌｉｎＤ１,１４例既表达ｃ     

细胞周期蛋白Cyclin D1 和CDK 4在新疆维吾尔族妇女宫颈癌中的表达及意义

目的:探讨Cyclin D1和CDK 4在新疆维吾尔族妇女宫颈癌发生、发展中的作用及相互关系.方法:采用免疫组化方法,检测100例宫颈癌组织和50例正常宫颈组织中Cyclin D1和CDK 4蛋白的表达情况.结果:宫颈癌组织中Cyclin D1 蛋白阳性表达率为75%,高于正常对照组的30%(P<0.01);CDK 4蛋白阳性表达率(87%),高于正常对照组(44%)(P<0.01).结论:Cyclin D1和CDK 4在新疆维吾尔族妇女宫颈癌中高表达,且与新疆维吾尔族妇女宫颈癌的发生、发展有关.     

大肠癌CDK4、Ki-67表达与Cyclin D1、p16蛋白表达的相关性及临床意义

目的　探讨CDK4、Ki 6 7蛋白表达与临床病理因素、预后的关系及其与CyclinD1、p16蛋白表达的关系。方法　采用sp法对 89例大肠癌标本进行CDK4、Ki 6 7、CyclinD1、p16蛋白免疫组化检测。结果　CDK4核表达在组织学分级Ⅲ级与Ⅰ、Ⅱ级之间的差异有显著性 (P <0 .0 5 ) ,与术后三年生存率未见明显相关 ,与CyclinD1、p16蛋白表达明显相关 (P <0 .0 5 )。Ki 6 7强阳性表达在组织学分级Ⅲ级与Ⅰ、Ⅱ级之间及临床分期D期与B、C期之间的差异有显著性 (P <0 .0 5 ) ,与术后三年生存率及CyclinD1表达明显相关 (P <0 .0 5 ) ,与p16蛋白表达无明显相关。CDK4核表达组Ki 6 7强阳性表达明显增高。结论　大肠癌CDK4核表达具有癌基因蛋白的特性 ,对组织分化及细胞增殖有一定的影响 ,Ki 6 7强阳性表达对评估预后、组织分化、临床进展有一定意义。CyclinD1、CDK4与p16共同参与大肠癌的发生与发展。     

Expression Analysis of Two Cancer-testis Genes,FBXO39 and TDRD4, in Breast Cancer Tissues and Cell Lines

Breast cancer accounts for one third of new cancer cases among women. The need for biomarkers for earlydetection is the stimulus to researchers to evaluate altered expression of genes in tumours. Cancer-testis (CT)genes are a group with limited expression in normal tissues except testis but up-regulation in a wide varietyof cancers. We here evaluated expression of two CT genes named FBXO39 and TDRD4 in 32 invasive ductalcarcinoma samples, 10 fibroadenomas and 6 normal breast tissue samples, in addition to two breast cancer celllines, MCF-7 and MDA-MB-231, by the means of quantitative real time RT-PCR. FBXO39 showed significantup-regulation in invasive ductal carcinoma samples in comparison with normal samples. It also was expressedin both cell lines and after RHOXF1 gene knock down it was down-regulated in MCF-7 but up-regulated in theMDA-MB-231 cell line. TDRD4 was not expressed in the MCF-7 cell line and any of the tissue samples excepttestis. However, it was expressed in MDA-MB-231 and was up-regulated after RHOXF1 gene knock down. Ourresults show that FBXO39 but not TDRD4 can be used for cancer detection and if proved to be immunogenic,might be a putative candidate for breast cancer immunotherapy.     

Glucose-6-phosphate dehydrogenase and NADPH oxidase 4 control STAT3 activity in melanoma cells through a pathway involving reactive oxygen species,c-SRC and SHP2

Background: Glucose-6-phosphate dehydrogenase (G6PD) participates in glucose utilization by catalysing the first step of the pentose-phosphate pathway in mammalian cells. Previous studies have shown that changes in G6PD levels can promote tumor cell proliferation or apoptosis via the STAT3/5 pathway in a human melanoma xenograft model. G6PD cooperates with NADPH oxidase 4 (NOX4) in the cellular metabolism of reactive oxygen species (ROS) and in maintaining the intracellular redox state. Methods: In this study, the effect of G6PD or NOX4 silencing in the melanoma line A375 was examined in terms of redox state, proto-oncogene tyrosine-protein kinase Src (c-Src) and the tyrosine-specific protein phosphatase SHP2 expression as well as cell cycle progression. Results: The results demonstrate that: (1) Downregulation of cyclin D1 and CDK4 and up-regulation of p53 and p21 occurred in response to silencing of G6PD and NOX4 thus resulting in G1/S cell cycle arrest and inhibition of A375 cell proliferation. (2) The blockade of cell proliferation is primarily due to a reduced DNA-binding activity of STAT3. (3) The DNA-binding activity of STAT3 was regulated by the upstream factors, c-SRC and SHP2. Silencing of NOX4 in A375 cells inhibited c-SRC and SHP2 regulated STAT3 activity. Conclusion: The data are consistent with a novel G6PD-NOX4-NADPH-ROS-c-SRC/SHP2 pathway controlling STAT3 activity in A375 melanoma cells.     

细胞周期蛋白CyelinD1和CDK4在新疆维吾尔族妇女宫颈癌中的表达及意义

目的：探讨CyclinD1和CDK4在新疆维吾尔族妇女宫颈癌发生、发展中的作用及相互关系。方法：采用免疫组化方法,检测100例宫颈癌组织和50例正常宫颈组织中CyclinD1和CDK4蛋白的表达情况。结果：宫颈癌组织中CyclinD1蛋白阳性表达率为75％,高于正常对照组的30％（P〈0．01）;CDK4蛋白阳性表达率（87％）,高于正常对照组（44％）（P〈0．01）。结论：CyelinD1和CDK4在新疆维吾尔族妇女宫颈癌中高表达,且与新疆维吾尔族妇女宫颈癌的发生、发展有关。     

Regulation of the G1/S phase of the cell cycle and alterations in the RB pathway in human lung cancer

The retinoblastoma (RB)–Cyclin (CCN)D1–p16 cell cycle pathway has a crucial role in lung tumorigenesis. Impairment of the RB pathway has been shown to occur in almost all lung tumors. A deregulation at any level of this core RB pathway seems to make cells insensitive to the mitogenic signaling that is required for cell cycle progression. To date, almost all participants in this pathway have been shown to be altered to a various degree in lung tumors. Some of the alterations are mutually exclusive, including RB and p16^INK4A. In small cell lung cancer, the RB tumor suppressor gene is inactivated in almost 90% of the tumors, whereas in non-small cell lung cancer, the cyclin-dependent kinase (CDK)4 inhibitor p16^INK4A is inactivated in 40–60% of the tumors. Many mechanisms may be responsible for activating the RB–Cyclin D1 pathway, including activating (CDK4) and inactivating mutations (p16^INK4A), deletions (RB and p16^INK4A), amplifications (CCND1 and CDK4), silencing methylation (p16^INK4A and RB), and hyper-phosphorylation (RB). As some of these alterations, such as p16^INK4A methylation, can also be detected in bronchial lavage and serum, they could potentially serve as useful markers for the early detection of lung cancer. This review summarizes recent experiments describing the variable roles of key-player molecules of the RB pathway and different mechanisms by which the RB pathway can be altered in lung cancer.     

EXPRESSION OF CYCLIN D1 AND CDK4 IN OSTEOSARCOMA OF THE JAWS

The main cause of the cellular malignant proliferation is the disorder of cell division and growth. The alterations of proteins and genes that involved in this process correlate closely with the occurrence and vicious phenotype of neoplasm. Cyclin D1 and cyclin-dependent kinase 4 (CDK4) regulatory proteins in G1 phase have a direct bearing on carcinogenesis. Presently, gene amplification, overexpression, chromosome inversion andtranslocation of cyclin D1 and CDK4 have been discovered in ma…     

Phase I study of PD 0332991, a cyclin-dependent kinase inhibitor, administered in 3-week cycles (Schedule 2/1)

Background:

This phase I, open-label, first-in-human study determined dose-limiting toxicities (DLTs) and maximum tolerated dose (MTD) of PD 0332991, an oral cyclin-dependent kinase 4/6 inhibitor with potent anti-proliferative activity in vitro/vivo.

Methods:

A total of 33 patients with retinoblastoma protein-positive advanced solid tumours or non-Hodgkin''s lymphoma refractory to standard therapy or for which no therapy was available received PD 0332991 once daily (QD) for 14 days followed by 7 days off treatment (21-day cycles; Schedule 2/1).

Results:

Six patients had DLTs (18% four receiving 200 mg QD; two receiving 225 mg QD); the MTD was 200 mg QD. Treatment-related, non-haematological adverse events occurred in 29 patients (88%) during cycle 1 and 27 patients (82%) thereafter. Adverse events were generally mild–moderate. Of 31 evaluable patients, one with testicular cancer achieved a partial response; nine had stable disease (⩾10 cycles in three cases). PD 0332991 was slowly absorbed (mean T_max 4.2 h) and eliminated (mean half-life 26.7 h). Volume of distribution was large (mean 3241 l) with dose-proportional exposure. Using a maximum effective concentration model, neutropenia was proportional to exposure.

Conclusion:

PD 0332991 was generally well tolerated, with DLTs related mainly to myelosuppression. The MTD, 200 mg QD, is recommended for phase II study.     

细胞周期调控相关蛋白Cyclin D1、CDK 4和pRb在新疆维吾尔族妇女宫颈癌中的表达及意义

 目的 探讨细胞周期调控相关蛋白Cyclin D1、CDK 4和pRb磷酸化状态在新疆维吾尔族妇女宫颈癌（简称维族）中表达及其意义。方法 应用免疫组织化学方法检测64例维族宫颈癌及43例维族正常宫颈组织中Cyclin D1、CDK 4表达和pRb磷酸化状态。结果 Cyclin D1和CDK4在宫颈癌组中阳性率分别为70%、87%,与正常宫颈组相比,差异有统计学意义（P<0.05）,pRb磷酸化在两组中差异有统计学意义（P<0.01）。结论 Cyclin D1 与CDK4 蛋白的高表达及pRb高磷酸化可能与维族宫颈癌发生发展有关。     

Recent advances with cyclin-dependent kinase inhibitors: therapeutic agents for breast cancer and their role in immuno-oncology

Introduction: Collaborative interactions between several diverse biological processes govern the onset and progression of breast cancer. These processes include alterations in cellular metabolism, anti-tumor immune responses, DNA damage repair, proliferation, anti-apoptotic signals, autophagy, epithelial-mesenchymal transition, components of the non-coding genome or onco-mIRs, cancer stem cells and cellular invasiveness. The last two decades have revealed that each of these processes are also directly regulated by a component of the cell cycle apparatus, cyclin D1.

Area covered: The current review is provided to update recent developments in the clinical application of cyclin/CDK inhibitors to breast cancer with a focus on the anti-tumor immune response.

Expert opinion: The cyclin D1 gene encodes the regulatory subunit of a proline-directed serine-threonine kinase that phosphorylates several substrates. CDKs possess phosphorylation site selectivity, with the phosphate-acceptor residue preceding a proline. Several important proteins are substrates including all three retinoblastoma proteins, NRF1, GCN5, and FOXM1. Over 280 cyclin D3/CDK6 substrates have b\een identified. Given the diversity of substrates for cyclin/CDKs, and the altered thresholds for substrate phosphorylation that occurs during the cell cycle, it is exciting that small molecular inhibitors targeting cyclin D/CDK activity have encouraging results in specific tumors.     

Mechanisms of therapeutic CDK4/6 inhibition in breast cancer

Cyclin dependent kinase (CDK) 4/6 inhibitors have advanced the treatment of metastatic breast cancer by targeting the cell cycle machinery, interrupting intracellular and mitogenic hormone signals that stimulate proliferation of malignant cells. Preclinical evidence demonstrated that derangements of cyclin D1, CDK4/6, and retinoblastoma expression are common in breast cancer, and suggested a therapeutic benefit from interrupting this axis required for cell cycle progression. Studies of cell lines and animal models of breast cancer have demonstrated the complex interplay between the cell cycle and estrogen receptor and human epidermal growth receptor 2 signaling, which informs our understanding of synergistic use of CDK4/6 inhibitors with endocrine therapy, as well as mechanisms of resistance to endocrine therapy. Interestingly, estrogen receptor activity leads to upregulation of cyclin D1 expression, but the estrogen receptor is also in turn activated by cyclin D1, independent of estrogen binding. Early CDK inhibitors were nonspecific and limited by systemic toxicities, while the current generation of CDK4/6 inhibitors have shown promise in the treatment of hormone receptor-positive breast cancer. Preclinical investigations of the three CDK4/6 inhibitors approved by the US Food and Drug Administration (palbociclib, ribociclib, and abemaciclib) lend further insight into their mechanism of action, which will hopefully inform the future use and refinement of these therapies. Finally, we summarize evidence for additional novel CDK4/6 inhibitors currently in development.     

骨肉瘤中Rb、p16、CDK4、cyclinD1蛋白的表达及其意义

目的 探讨骨肉瘤中Ｒｂ、ｐ１６、ＣＤＫ４、ｃｙｃｌｉｎＤ１的蛋白表达、相互关系及其意义。方法 应用免疫组化方法对４０例骨肉瘤组织中Ｒｂ、ｐ１６、ＣＤＫ、ｃｙｃｌｉｎＤ１的蛋白表达状况进行检测。结果 ４０例骨肉瘤Ｒｂ、ｐ１６蛋白表达缺失分别为７２．５％（２９／４０和）２２．５％（９／４０）;１１例ｐＲｂ阳性的骨肉瘤中的８例ｐ１６阴性,２９例ｐＲｂ阴性的骨肉瘤中２８例ｐ１６阳性;５２．５％（２１／４０     

视黄酸依赖反义cyclin D1对HL-60细胞细胞周期素依赖性激酶的影响

目的探讨视黄酸(retinoic acid,RA)及其诱导的反义(anti-sense,AS)cyclin D1对HL-60细胞的细胞周期素激酶CDK4的影响.方法通过构建含有视黄酸反应元件(retinoic acid response element, RARE)的反义cyclin D1 RNA表达载体pCI-neo/RARE3-TK/Ascyclin D1,使反义cyclin D1的表达可受视黄酸诱导调控.以脂质体转染HL-60白血病细胞后,运用MTT比色法检测细胞增殖功能、NBT还原实验观测细胞分化状况,RT-PCR以及western-blot等方法观测视黄酸处理后CDK4的mRNA和蛋白表达水平的改变.结果 RA处理后,细胞生长显著减慢,分化能力明显增强,CDK4 mRNA和蛋白表达水平均有所降低,其中,以反义cyclin D1转染细胞经RA处理后变化更为显著.结论 RA及其诱导的反义cyclin D1对HL-60细胞的抑制增殖与诱导分化效应可能与CDK4表达下调有关.     

米非司酮对裸鼠子宫内膜癌细胞HHUA移植瘤生长及COX-2、CDK4的影响

背景与目的子宫内膜癌是女性常见的恶性肿瘤,内膜癌中存在COX-2,CDK4的表达,并与肿瘤的发生发展有关,近年来发现米非司酮有抗肿瘤作用,但其作用机制不十分明了,我们研究米非司酮(mifeprisitone,MIF)对人子宫内膜癌HHUA细胞株裸鼠移植瘤生长的抑制作用及对COX-2,CDK4的影响,以确定米非司酮能否通过干预肿瘤中COX-2,CDK4的表达而抑制肿瘤生长。方法体外培养人子宫内膜癌HHUA细胞,裸鼠皮下接种细胞建立裸鼠移植瘤动物模型,随机将10只移植瘤荷瘤裸鼠分为两组,MIF组采用灌胃法[50mg/(kg·d)],对照组灌等量溶剂。观察治疗前后移植瘤的体积变化。免疫组织化学染色法检测移植瘤组织COX-2、CDK4的表达,后采用Leica IM50免疫组织化学评分软件(HSCORE)对染色结果进行半定量分析,HE染色观察肿瘤组织的病理变化。结果治疗6周后,MIF组移植瘤体积(115.25±10.97)mm3,对照组移植瘤体积(313.25±43.92)mm3,两者相比,差异有非常显著性(P<0.01);HE染色见MIF组较对照组移植瘤坏死面积显著增加;MIF组COX-2HSCOREs为(78.2±11.3),对照组(205.9±26.7),两者相比,差异有显著性(P<0.01);MIF组CDK4HSCOREs为(113.4±18.2),对照组HSCOREs为(238.7±35.9),两者相比,差异有非常显著性(P<0.01)。结论MIF具有抑制人子宫内膜癌细胞HHUA裸鼠移植瘤生长的作用;其机制可能与下调COX-2、CDK4的表达有关。