异丹叶大黄素下调膀胱癌细胞的细胞周期蛋白D1表达并诱导G0/G1细胞周期阻滞

 目的：探索异丹叶大黄素（ISO）抑制膀胱癌细胞侵袭、转移和增殖活性的机制。方法：以ISO作用于人源性膀胱癌UMUC3细胞后,倒置相差显微镜下观察并以ATP生物荧光法检测癌细胞增殖活性;以RT-PCR和Western blotting法检测细胞周期蛋白D1表达;以流式细胞术检测细胞周期的变化;细胞划痕法检测细胞迁移活性。结果：20 μmol/L浓度以上ISO可显著抑制UMUC3细胞的增殖,其IC50为(22.5±2.8) μmol/L。同时 UMUC3细胞的细胞周期蛋白D1 mRNA和蛋白水平均显著降低。以低浓度5 μmol/L ISO预处理UMUC3细胞后,与对照组（47.33%）进行比较,可分别在12 h（58.82％）和24 h（63.94％）显著诱导细胞G0/G1期阻滞（P<0.01）。细胞划痕法检测证实5 μmol/L ISO可显著抑制膀胱癌细胞的迁移活性。结论：ISO可抑制膀胱癌细胞增殖,下调细胞周期蛋白D1表达,诱导G0/G1细胞周期阻滞,抑制细胞的迁移能力。     

异丹叶大黄素抑制HepG2细胞生长及诱导其凋亡的实验研究

目的通过异丹叶大黄素抑制HepG2细胞的生长及诱导其凋亡,探讨异丹叶大黄素抗肿瘤作用的分子机制。方法我们采用FACS、Hochest33258、MTT等方法来检测HepG2细胞生长及凋亡的情况。结果异丹叶大黄素药物组的细胞凋亡明显大于对照组,药物组的细胞生长情况明显受到抑制,P<0.05,且还呈浓度依赖和时间依赖性。结论异丹叶大黄素药物可抑制Hep62细胞的增殖及诱导的HepG2细胞凋亡,表明异丹叶大黄素药物具有抗肿瘤的药理作用。     

软组织平滑肌肉瘤中细胞周期蛋白D1基因扩增分析及蛋白表达检测

我们采用差异聚合酶链反应 (ｄｉｆｆｅｒｅｎｔｉａｌＰＣＲ ,ＤＰＣＲ)检测软组织平滑肌肉瘤 (ＬＭＳ)中细胞周期蛋白Ｄ1(ｃｙｃｌｉｎＤ1)基因ＣＣＮＤ1的扩增情况 ,并应用免疫组织化学方法检测ｃｙｃｌｉｎＤ1表达 ,以期了解二者的关系。一、材料与方法1.标本 :38例ＬＭＳ、10例平滑肌瘤和 10例正常平滑肌组织取自本院病理科 1989～ 1998年存档蜡块 ,肿瘤发生部位包括躯干、四肢、腹腔和腹膜后 (胃肠道除外 )。2 .ＤＰＣＲ :多巴胺受体基因 (ｄｏｐａｍｉｎｅｒｅｃｅｐｔｏｒＤ2 ,ＤＲＤ2 )与ＣＣＮＤ1基因同位于 11号染色体的长臂上 (…     

喉鳞状细胞癌患者视网膜母细胞瘤蛋白和细胞周期调节蛋白D1异常表达

目的:探讨喉鳞状细胞癌(LSCC)患者视网膜母细胞瘤蛋白(pRb)和细胞周期调节蛋白D1(Cyclin D1)的异常表达及其相关性。方法:选择36例喉部疾病手术患者,包括LSCC(LSCC组)、声带息肉(息肉组)和癌前病变(癌前组)各12例,各组均于手术后取病变组织,采用Western-blotting检测pRb和Cyclin D1蛋白表达水平,统计学分析各组两指标差异及相关性。结果:与息肉组比较,癌前组pRb表达水平降低(P0.01),Cyclin D1表达水平升高(P0.01);与癌前组比较,LSCC组pRb水平更低(P0.01),Cyclin D1水平更高(P0.01)。三组pRb与Cyclin D1蛋白水平均呈显著负相关(r分别为-0.94、-0.83和-0.92,P均0.01)。结论:LSCC的发生和发展与pRb表达降低和Cyclin D1表达增加有关。     

p15、细胞周期蛋白D1和转移生长因子β1在非小细胞肺癌中的表达及其意义

一、材料与方法1.材料 :实验材料取自内蒙古医学院病理教研室在1996～ 1998年间的病理组织存档蜡块 ,共 16 5份。按照 1980年ＷＨＯ肺癌分类标准进行组织学分类 ,并均经病理证实。其中鳞癌 80例 ,腺癌 49例 ,不典型增生 19例 ,鳞状上皮化生17例 ;在 12 9例非小细胞肺癌中 ,同一病例手术切取标本存档蜡块不仅有肺癌组织蜡块也有淋巴结组织蜡块 ;其中淋巴结组织经ＨＥ染色证实有淋巴结转移的 35例 ,无淋巴结转移的 2 8例。其余 6 6例中 ,或同一病例手术切取标本存档蜡块仅有肺癌组织而无淋巴结组织 ,或同一病例标本存档蜡块中虽有淋巴结组织 ,…     

双肾上腺皮质激素样激酶-1与细胞周期蛋白D1在结直肠癌组织中的表达及临床意义

目的:探讨双肾上腺皮质激素样激酶-1(DCLK1)与细胞周期蛋白D1(cyclin D1)在结直肠癌组织中的表达及其临床意义。方法:采用免疫组织化学S-ABC法检测59例结直肠癌组织和28例结直肠良性病变组织中DCLK1及cyclin D1的表达及分布。结果:DCLK1及cyclin D1在结直肠癌组织中的表达明显高于结直肠良性病变组织,平均光密度值(AOD)为0.423±0.021及0.386±0.042;而在结直肠良性病变组织中其阳性反应的平均光密度值为0.267±0.053及0.211±0.026,差异具有统计学意义。DCLK1及cyclin D1的阳性表达与患者的年龄、性别、肿瘤分化程度和DUCKs分期无关,而与组织良性或恶性、肿瘤的大小及淋巴结转移相关。结论:结直肠癌组织中DCLK1及cyclin D1的高表达可能与结直肠癌的发生、发展、转移相关,可作为结直肠癌转移及预后监测的一项重要指标。     

套细胞淋巴瘤的临床病理特征和细胞周期蛋白D1在诊断中的意义

目的 观察套细胞淋巴瘤的临床病理学特征及细胞周期蛋白D1染色在诊断中的意义。方法 对8例淋巴结套细胞淋巴瘤作临床病理观察及随访,LSAB法做免疫表型分析（CD45RO、CD5、CD20、细胞周期蛋白D1、Ki-67、bcl-2）。结果 患者年龄43-78岁（平均年龄57岁）,男女3：1。组织学特点为：（1）淋巴结结构破坏并被单一的淋巴样细胞所取代,淋巴细胞以套区增生性、结节性、弥漫性三种模式增生。（2）淋巴样细胞核有一定的不规则性,染色质中等致密,核分裂象少见,类似中心细胞。其中有3例转变为高度侵袭性的母细胞样变型。所有的病例都呈cyclinD1与bcl-2阳性、CD20阳性、CD45RO阴性、CD5阳性。结论 套细胞淋巴瘤有其特征的形态改变及免疫表型。根据组织病理学特征及cyclin D1阳性,可与其它类型的小B细胞淋巴瘤相鉴别。套细胞淋巴瘤的母细胞样变型也应当与其它变型区别。     

β-连环蛋白、细胞周期蛋白D1和c-myc表达与胰腺癌发生及生物学行为的关系

目的 研究β-连环蛋白(β-cat)的异常表达、细胞周期蛋白D1(cyclin D1)和 c-myc的高表达与胰腺癌发生、增殖、浸润、转移和预后的关系。方法 应用免疫组织化学PicTure~(TM)二步法,检测47例胰腺癌组织、12例胰腺导管上皮内肿瘤(上皮中度不典型增生)和10例正常胰腺组织中β-cat、cyclin D1和c-myc的表达。同时检测胰腺癌增殖细胞核抗原(PCNA),作为胰腺癌增殖状态的指标。结果 β-cat在10例正常胰腺组织均呈正常表达,而cyclin D1和c-myc均呈阴性。三者在胰腺导管上皮内肿瘤和胰腺癌中的表达率差异无显著性[分别为6/12和68.1％(3/47)、6/12和74.5％(35/47)、5/12和70.2％(33/47),均P>0.05]。β-cat异常表达率与胰腺癌的转移和1年生存率显著相关(均P<0.05),而与胰腺癌大小、分化程度、增殖活性及浸润无关(均P>0.05)。cyclin D1的表达率与胰腺癌的增殖和分化程度有关(均P0.05)。c-myc的表达率与胰腺癌的大小、分化程度、浸润、转移和1年生存率无关(均P>0.05),而与胰腺癌组织的增殖活性密切有关(P<0.05)。β-cat异常表达与cyclin D1和c-myc的高表达在胰腺导管上皮内肿瘤和胰腺癌中均有显著的正相关性(均P<0.05,r=1.000、0.845、0.437、0.452)。结论 β-cat     

细胞周期蛋白D1、p21WAF1蛋白和增殖细胞核抗原在乳腺癌的表达及与淋巴结转移和预后的关系

我们在以往研究的基础上，检测了乳腺癌组织中细胞周期蛋白(cyclin)D1、p21^WAF1蛋白及增殖细胞核抗原(PCNA)的表达以及有cyclin D1过表达乳腺癌淋巴结转移灶中cyclin D1的表达，以探讨其特别是cyclin D1在乳腺癌发生发展中的作用及其与淋巴结转移关系和预后意义。     

miR-134-3p靶向细胞周期蛋白D1抑制胃癌细胞增殖的作用及机制

目的：研究mi R-134-3p靶向细胞周期蛋白D1(CCND1)抑制胃癌细胞增殖的作用及机制。方法：收集手术切除的胃癌组织及癌旁组织,培养正常胃黏膜上皮细胞株GES-1及胃癌细胞株MGC-803、SGC-7901、BGC-823,检测mi R-134-3p、CCND1的表达水平;BGC-823细胞分组并转染阴性对照(NC)模拟物或mi R-134-3p模拟物、感染NC腺病毒或CCND1腺病毒,检测细胞增殖抑制率、细胞周期比例、mi R-134-3p及CCND1表达水平;双荧光素酶报告基因验证mi R-134-3p与CCND1的靶向结合;饲养BALB/c裸鼠,皮下注射感染NC腺病毒或mi R-134-3p腺病毒的BGC-823,成瘤后测定移植瘤质量、体积及CCND1表达水平。结果：与癌旁组织相比,胃癌组织中mi R-134-3p表达水平明显降低、CCND1表达水平明显升高(P<0.05),且mi R-134-3p与CCND1呈负相关;与GES-1细胞相比,BGC-823、MGC-803、SGC-7901细胞中mi R-134-3p表达水平明显降低,CCND1表达水平明显升高(P&...     

碱性成纤维细胞生长因子对卵巢癌CAOV3细胞cyclin D1及GADDl53表达的影响

目的:研究碱性成纤维细胞生长因子(bFGF)对人卵巢癌CAOV3细胞细胞周期调节蛋白cyclinD1及GADD153表达的影响,探讨bFGF促进人卵巢癌CAOV3细胞增殖、抑制凋亡的信号机制。方法:利用无血清饥饿诱导卵巢癌CAOV3细胞凋亡。分为对照组、bFGF组。分别应用MTT、流式细胞术、琼脂糖凝胶电泳观察25、50、75μg/L bFGF对CAOV3细胞增殖率、细胞周期及细胞凋亡的影响。利用Western blotting检测bFGF对CA-OV3细胞cyclin D1、GADD153以及转录因子(c-Fos、c-Jun)表达的影响。结果:与对照组相比,bFGF呈剂量依赖性加速CAOV3细胞细胞周期进程,促进细胞增殖,抑制饥饿诱导的凋亡(P<0.01);呈时间依赖性促进cyclinD1、c-Fos、c-Jun,抑制GADD153蛋白表达(P<0.01)。结论:bFGF可能通过上调cyclin D1、c-Fos、c-Jun,下调GADD153表达促进细胞增殖,抑制饥饿诱导的卵巢癌CAOV3细胞凋亡。     

碱性成纤维细胞生长因子对卵巢癌CAOV3细胞cyclin D1及GADD153表达的影响

目的：研究碱性成纤维细胞生长因子（bFGF）对人卵巢癌CAOV3细胞细胞周期调节蛋白cyclin D1 及GADD153表达的影响，探讨bFGF促进人卵巢癌CAOV3细胞增殖、抑制凋亡的信号机制。方法:利用无血清饥饿诱导卵巢癌CAOV3细胞凋亡。分为对照组、bFGF组。分别应用MTT、流式细胞术、琼脂糖凝胶电泳观察 25、50、75 μg/L bFGF对CAOV3细胞增殖率、细胞周期及细胞凋亡的影响。利用Western blotting检测bFGF对CAOV3细胞cyclin D1、GADD153以及转录因子（c-Fos、c-Jun）表达的影响。结果:与对照组相比，bFGF呈剂量依赖性加速CAOV3细胞细胞周期进程，促进细胞增殖，抑制饥饿诱导的凋亡（P＜0.01）；呈时间依赖性促进cyclin D1、c-Fos、c-Jun，抑制GADD153蛋白表达（P＜0.01）。结论:bFGF可能通过上调cyclin D1、c-Fos、c-Jun，下调GADD153表达促进细胞增殖，抑制饥饿诱导的卵巢癌CAOV3细胞凋亡。     

Upregulation of ATF-3 is correlated with prognosis and proliferation of laryngeal cancer by regulating Cyclin D1 expression

Objective: This study aimed to investigate the expression and significance of ATF-3 in laryngeal squamous cell carcinoma (LSCC). Methods: Expression of ATF-3 was examined using immunohistochemistry methods in samples from 83 cases of LSCC carcinoma. MTT assay was used to detect proliferation of Hep-2 cells after ATF-3 knocked down by siRNA lentivirus. A mouse model was used to investigate the inhibitive role of ATF-3 siRNA in LSCC xenografts. Realtime RCR was used to detect Cyclin D1 expression after ATF-3 downregulation in Hep-2 cells. Results: The expression of ATF-3 was positively detected in all the 83 cases of LSCC cancer tissues while Only 4 cases of adjacent non-neoplastic tissues were detected with positive ATF-3 expression. The ATF-3 expression was statistically related with T stage, neck nodal metastasis, clinical stage and prognosis of LSCC. Both cell proliferation in vitro and tumor growth in vivo were suppressed after ATF-3 knockdown. Furthermore, the expression of Cyclin D1 was decreased after ATF-3 downregulation in Hep-2 cells. Conclusion: ATF-3 is involved in the progress of LSCC, and may provide clinical information for evaluation of prognosis of LSCC. The oncologic role of ATF-3 may be correlated with Cyclin D1 regulation.     

Cyclin D1 expression in prostate carcinoma

The purpose of this study was to investigate the relationship between cyclin D1 expression and clinicopathological parameters in patients with prostate carcinoma. We assessed cyclin D1 expression by conventional immunohistochemistry in 85 patients who underwent radical prostatectomy for prostate carcinoma and 10 normal prostate tissue samples retrieved from autopsies. We measured nuclear immunostaining in the entire tumor area and based the results on the percentage of positive tumor cells. The preoperative prostate-specific antigen (PSA) level was 8.68±5.16 ng/mL (mean±SD). Cyclin D1 staining was positive (cyclin D1 expression in >5% of tumor cells) in 64 cases (75.4%) and negative (cyclin D1 expression in ≤5% of tumor cells) in 21 cases (including 15 cases with no immunostaining). Normal prostate tissues were negative for cyclin D1. Among patients with a high-grade Gleason score (≥7), 86% of patients demonstrated cyclin D1 immunostaining of >5% (P<0.05). In the crude analysis of cyclin D1 expression, the high-grade Gleason score group showed a mean expression of 39.6%, compared to 26.9% in the low-grade Gleason score group (P<0.05). Perineural invasion tended to be associated with cyclin D1 expression (P=0.07), whereas cyclin D1 expression was not associated with PSA levels or other parameters. Our results suggest that high cyclin D1 expression could be a potential marker for tumor aggressiveness.     

The influence of arsenic trioxide on the cell cycle,apoptosis and expression of cyclin D1 in the Jurkat cell line

Cyclin D1 drives cell cycle progression at the G1/S transition and is believed to play a significant role in tumorigenesis, contributing to efficient proliferation of many cancer cells. Consequently, it is also recognized as an end-point biomarker of therapeutic outcome for different treatment modalities in cancer. In this study we aimed to evaluate the expression and localization of cyclin D1 in arsenic trioxide (ATO) treated Jurkat cells (lymphoblastic leukemia cell line) and to correlate these results with the extent of cell death and/or cell cycle alterations. Jurkat cells were incubated with increasing concentrations of ATO (0.2, 0.6 and 1.0 μM) for 24 h in standard cell culture conditions. To reach our goal we performed annexin V/PI labeling for detection of cell death and RNase/PI labeling for evaluation of cell cycle distribution, which were followed by the respective flow cytometric analyses of ATO-treated Jurkat cells. Transmission electron microscopy was applied for visualization of the cell ultrastructure. For cyclin D1 estimation a biparametric cyclinD1/cell cycle assay was done and localization of the protein was shown after immuno-labeling using light microscopy (ABC procedure) and confocal fluorescence microscopy. We found that there were no significant changes in the percentages of cyclin D1-positive cells after the treatment with ATO, but at the same time mean fluorescence intensity reflecting cyclin D1 content was gradually increasing along with the cell cycle progression, irrespective of the applied dose of the drug. On the other hand, we found a nuclear-cytoplasmic shift of this protein as a major treatment-related response, which was in good accord with an increased rate of cell death and suggested that cyclin D1 cytoplasmic degradation is an important determinant of the therapeutic efficiency of ATO in the Jurkat cell line.     

Cyclin D1 expression by histiocytes may mimic cyclin D1-positive proliferation centres of chronic lymphocytic leukaemia/small lymphocytic lymphoma

Aims

Cyclin D1, generally considered to be absent in chronic lymphocytic leukaemia/small lymphocytic lymphoma (CLL/SLL), has been reported in the proliferation centres (PCs) of recent CLL/SLL cases. Cyclin D1 immunostaining in CLL/SLL may lead to diagnostic confusion. The objective of this study was to identify the types of stained cells and the impact on diagnosis.