Cyclin D1在黄芪甲苷诱导BMSCs向神经元样细胞分化中的表达

目的:研究黄芪甲苷诱导骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)分化的特点和诱导过程中Cyclin D1的表达特点。方法:采用100 mg·L^(-1)黄芪甲苷诱导BMSCs分化,在1、12、24 h倒置显微镜观察细胞形态变化,免疫细胞化学方法观察巢蛋白(Nestin)、神经元特异性烯醇化酶(neuron specific enolase,NSE)、Cyclin D1的表达,RT-PCR检测Cyclin D1 mRNA的表达。结果:收集获得的骨髓细胞采用原代和传代培养,至第3代BMSCs细胞基本纯化。经100 mg·L^(-1)黄芪甲苷作用后,部分细胞自胞体长出突起,形态似神经元样细胞且表达Nestin和NSE抗体,Nestin的表达时相早于NSE且表达强度高于NSE(P<0.05)。在药物作用1 h后,BMSCs细胞Cyclin D1不论在基因转录水平还是在蛋白表达水平均出现大幅度下调,此时BMSCs开始有Nestin的表达;药物作用12、24 h,BMSCs细胞Cyclin D1表达与药物作用前表达水平无显著性差异(P>0.05)。结论:黄芪甲苷可能首先通过调整细胞周期,延长了G1期向S期转变的时间,从而启动了BMSCs的分化程序,诱导BMSCs向神经干细胞分化。在黄芪甲苷的营养作用下,细胞的突起进一步生长并向神经元分化。

子宫内膜增生组织中cyclinD1、PCNA和MMP-9表达水平与病理特征的关系

目的探究子宫内膜增生组织中G1/S-特异性周期蛋白-D1(cyclinD1)、增殖细胞核抗原(PCNA)和基质金属蛋白酶9(MMP-9)表达水平与病理特征的关系。方法回顾性分析2018年2月至2023年3月期间阜南县中医院收治的108例子宫内膜增生患者作为研究对象,检测患者子宫内膜增生组织中cyclinD1、PCNA和MMP-9表达水平;同时采用病理HE切片评估患者增生组织的病理特征。采用Spearman相关性分析探究cyclinD1、PCNA和MMP-9水平与病理特征之间的关联性。结果108例患者中,单纯性增生患者73例(67.59%),复杂性增生患者24例(22.22%),非典型增生患者11例(10.19%)。复杂性增生患者cyclinD1、PCNA和MMP-9阳性表达的病例数占比高于单纯性增生患者;非典型增生患者cyclinD1、PCNA和MMP-9阳性表达的病例数占比显著高于单纯性增生患者和复杂性增生患者,差异具有统计学意义(P<0.05)。Spearman相关性分析结果显示,cyclinD1、PCNA及MMP-9的表达水平与子宫内膜增生组织的病理特征存在关联性。结论子宫内膜增生组织的cyclinD1、PCNA及MMP-9水平与病理特征存在密切关联,上述指标的高水平表达提示患者存在较高的癌前病变风险。

cyclin D2、Bcl-2在弥漫大B细胞淋巴瘤中的表达意义

目的研究细胞周期蛋白D2(cyclin D2)、B淋巴细胞瘤-2基因(Bcl-2)在弥漫大B细胞淋巴瘤(DLBCL)中的表达意义。方法采用回顾性分析,观察对象为2017年3月至2022年3月入东莞市人民医院就诊的80例DLBCL患者与20例淋巴组织反应性增生(RLH)患者,分别设定为研究组与对照组,两组患者均经术后病理检查确诊,术前未行任何治疗。选取免疫组织化学法测定两组cyclin D2、Bcl-2蛋白表达情况,对cyclin D2、Bcl-2表达之间的相互关系及与DLBCL患者病理特征相关性进行分析。结果研究组cyclin D2蛋白阳性率、Bcl-2蛋白阳性率分别为33.75%(27/80)、62.50%(50/80),对照组cyclin D2蛋白阳性率、Bcl-2蛋白阳性率分别为5.00%(1/20)、10.00%(2/20);研究组cyclin D2蛋白阳性率、Bcl-2蛋白阳性率较对照组更高,差异均有统计学意义(P<0.05)。cyclin D2蛋白表达、Bcl-2蛋白表达与DLBCL患者的免疫分型、Ann Arbor分期有明显相关性(P<0.05),与组织类型、肿瘤部位、性别及年龄无明显相关性(P>0.05)。DLBCL中cyclin D2与Bcl-2的表达呈正相关(r=1.000,P<0.05)。结论cyclin D2、Bcl-2在DLBCL中呈高表达,可能参与了DLBCL的发生及发展,两者具有协同作用,可辅助评价生物恶性程度。

Bcl-1 Rearrangement and Cyclin D1 Protein Expression in Multiple Myeloma Precursor Cells

The rearrangement of Bcl-1 gene (Bcl-1/IgH rearrangement) and expression of cyclin D1 in multiple myeloma (MM) precursor cells were studied and the role of cyclin D1 in the pathogenesis of MM was investigated. The BCL-1 rearrangement and cyclin D1 protein expression in 15 cases of MM were detected. By using hemi-nested polymerase chain reaction (PCR) the genomic DNA from fresh peripheral blood and bone marrow was amplified and the expression of cyclin D1 in the smears was detected by using immunohistochemical method. Ten volunteer with normal bone marrow served as control group. The results showed Bcl-1 rearrangement was detectable in 3/15 (20 %) MM patients and cyclin D, expression in 4/15 (27 % ) MM patients. BeLl-1 rearrangement and cyclin D1 protein expression were also detected in MM precursor cells. No overexpression of cyclin D1 or the rearrangement of the BeL-1 gene was found in the 10 volunteers. It was concluded that Bel-1 rearrangement and cyclin D1 protein overexpression were detected in MM precursor cells, speculating that overexpression of cyclin D1 protein may play an initial (critical) role in the pathogenesis of MM.

Effects of Serum from Aplastic Anemia patients on the Expression of Cyclin D3 Isoform in Umbilical Cord Blood CD34^+ Cells

Summary: The pathogenesis of aplastic anemia (AA) was explored and the effects of AA serum on the expression of crucial cyclin D isoform (cyclin D3) in umbilical cord blood hematopoietic stem/progenitor cells were observed. The CD34+ cells were isolated from the cord blood with MIDI-MACS Semi-solid methylcellulose culture technique was used to measure the formation of CFU-GM; The expression level of cyclin D3 was assayed by semi-quantitative RT-PCR and Western-blot after the hematopoietic stem/progenitor cells were incubated in AA serum. The results showed that the AA serum could inhibit the formation of CFU-GM and down regulate the expression level of the cyclin D3 at the mRNA and protein level respectively. In conclusion, the AA serum could inhibit the proliferation of hematopoietic stem cells and down regulate level of cyclin D3, which might be one mechanism of hematopoiesis inhibition in AA.

Molecular mechanism underlying the functional loss of cyclindependent kinase inhibitors p16 and p27 in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is one of the most common human cancers,and its incidence is still increasing in many countries.The prognosis of HCC patients remains poor,and identification of useful molecular prognostic markers is required.Many recent studies have shown that functional alterations of cell-cycle regulators can be observed in HCC.Among the various types of cell-cycle regulators,p16 and p27 are frequently inactivated in HCC and are considered to be potent tumor suppressors.p16,a G1-specific cell-cycle inhibitor that prevents the association of cyclindependent kinase(CDK)4 and CDK6 with cyclin D1,is frequently inactivated in HCC via CpG methylation of its promoter region.p16 may be involved in the early steps of hepatocarcinogenesis,since p16 gene methylation has been detected in subsets of pre-neoplastic liver cirrhosis patients.p27,a negative regulator of the G1-S phase transition through inhibition of the kinase activities of Cdk2/cyclin A and Cdk2/cyclin E complexes,is now considered to be an adverse prognostic factor in HCC.In some cases of HCC with increased cell proliferation,p27 is overexpressed but inactivated by sequestration into cyclin D1-CDK4-containing complexes.Since loss of p16 is closely related to functional inactivation of p27 in HCC, investigating both p16 and p27 may be useful for precise prognostic predictions in individuals with HCC.

Id2 regulates the proliferation of squamous cell carcinoma in vitro via the NF-κB/Cyclin D1 pathway

Squamous cell carcinoma (SCC) is a significant cause of cancer morbidity and mortality worldwide, with an incidence of up to 166 cases per 100 000 population. It arises in the skin, upper aerodigestive tract, lung, and cervix and affects more than 200 000 Americans each year. We report here that a microarray experiment comparing 41 SCC and 13 normal tissue specimens showed that Id2, a gene that controls the cell cycle, was significantly up-regulated in SCC. Enforced expression of Id2 in vitro stimulated the proliferation of SCC cells and up-regulated the transcription of nuclear factor kappa B (NF-κB) and cyclin D1. Enhancement of the NF-κB activity with p65 significantly increased the cell proliferation and the transcription of cyclin D1, whereas inhibition of the NF-κB activity with I kappa B alpha mutant (IκBα M) and pyrroline dithiocarbamate (PDTC) abrogated cell proliferation and transcription of cyclin D1. Furthermore, a mutated NF-κB binding site in the cyclin D1 promoter fully abrogated the Id2-induced transcription of cyclin D1. Taken together, these data indicate that Id2 induces SCC tumor growth and proliferation through the NF-κB/cyclin D1 pathway.

Inhibition of Proliferation Induced by Cyclin D1 Gene Silence in Human Renal Carcinoma ACHN Cells

Objective： To confirm feasibility of Cyclin D1 gene as a new target for cancer gene therapy and to verify the effectiveness of shRNA expression vector-mediated gene silencing. Methods： A RNA interference DNA template targeting Cyclin D1 gene was designed and synthesized. By ligation, the fragment was inserted into Pgenesil-1-U6 to constract the recombinant plasmid Pgenesil-shRNA-Cyclin D1. The identified recombinant plasmid was transfected into ACHN cells with lipofactamine. Cyclin D1 mRNA and protein expression was analyzed by RT-PCR and western-blotting. MTT method was used for observing cell proliferation and drawing growth curve. The cell cycle and ratios of apoptotic cell were assessed by flow cytometric detection. The ability of invasion of cell migration was detected by Transwell chamber invasive models. Results： The plasmid was constructed successfully. After interference, The expression rate of Cyclin D1 mRNA decreased to 0.10±0.04 in Cyclin D1-shRNA（experimental） group and were significantly lower than Pgenesil-NC （negative） group （0.92±0.03） and ACHN （blank control） group（0.94±0.04）（P0.05）. As well, the expression rate of Cyclin D1 protein was decreased evidently in experimental group. The results of flow cytometric detection showed that, including early and late apoptotic cells, the apoptotic ratio of experimental group increased to （37.26±0.60）% significantly, while, the negative group and blank control group were only （4.62±0.40）% and （5.95±1.20）%, respectively. The cell growth curves indicated that the proliferation of experimental group cells was inhibited significantly（P0.05） and Transwell results suggested that the abilities of invasion cells transfected with Pgenesil–CyclinD1-shRNA decreased conspicuously（P0.05）. Conclusion： The shRNA can inhibit Cyclin D1 expression, specifically and persistently. The down-regulation of Cyclin D1 expression can inhibit the proliferation and induce the apoptosis of renal cell adenocarcinoma cell line ACHN.

Effects of Cyclin D1 Antisense Oligodeoxyneucleotides on the Growth and Expression of G_1 Phase Regulators in Gastric Carcinoma Cells

To investigate the effects of Cyclin D1 antisense oligodeoxyneucleotides (ASODN) on the growth, cell cycle progression and expression of G 1 phase regulators in human gastric carcinoma cell lines SGC7901 and HS746T, phosphorothioate modified Cyclin D1 ASODN were encapsulated by LipofectAMINE2000 and transfected into gastric carcinoma cells. Dose dependent inhibitory effects were induced by Cyclin D1 ASODN in two gastric carcinoma cell lines. Treatment of gastric carcinoma cells with 0.2 μmol/L Cyclin D1 ASODN for 24 h could significantly inhibit their growth in vitro and in vivo , reduce expression of Cyclin D1mRNA to 26.3 % (SGC7901) and 17.3 % (HS746T) respectively. The percentage of cells in G 0/G 1 phase was increased as revealed by flow cytometry. Immunohistochemical staining showed that the expression of p21 was increased and the expression of Cyclin D1 and pRb was decreased in the two cell lines; the expression of p27 was increased in HS746T, but unchanged in SGC7901. Cyclin D1 ASODN could inhibit the growth and the expression of Cyclin D1 mRNA in gastric carcinoma cells, influence the cell cycle and expression of its regulators.

Vitamin C Inhibits Benzo[a]pyrene-Induced Cell Cycle Changes Partly via Cyclin D1/E2F Pathway in Human Embryo Lung Fibroblasts

Objective To study the molecular mechanism of the inhibitory effects of vitamin C on benzo[a]pyrene （B[a]P）-induced changes of cell cycle in human embryo lung fibroblast （HELF） cells. Methods The stable transfectants, HELF transfected with antisense cyclin D1 and antisense CDK4, were established. Cells were cultured and pretreated with vitamin C before stimulation with B[a]P for 24 h. The expression levels of cyclin DI, CDK4, E2FI, and E2F4 were determined by Western blot. Flow cytometric analysis was employed to detect the distributions of cell cycle. Results B[a]P significantly elevated the expression levels of cyclin D 1, E2F1, and E2F4 in HELF cells. Vitamin C decreased the expression levels of cyclin D 1, E2F1, and E2F4 in B [a]P-stimulated HELF cells. Dose-dependent relationships were not found between the different concentrations of vitamin C （10, 100, 500, 1000, and 5000 lamol/L） and the expression levels of cyclin D 1, E2F1, and E2F4 in HELF cells. The expression levels of cyclin D1, E2FI, and E2F4 in B[a]P-treated transfectants were lower than those in B[a]P-treated HELF cells. The expression levels of cyclin DI and E2F4 treated with vitamin C and antisense cyclin D1 were decreased compared with those treated with antisense cyclin DI alone. The effects of vitamin C combined with antisense CDK4 on the expression levels of cyclin DI and E2FI/E2F4 were similar to those of antisense CDK4 alone. B[a]P progressed HELF cells from GI to S phase. Both vitamin C and antisense cyclin DI suppressed the changes of cell cycle progressed by B[a]P. However, antisense CDK4 did not attenuate the above changes. Vitamin C combined with antisense CDK4 markedly suppressed B[a]P-induced changes of cell cycle as compared with antisense CDK4. But the inhibitory effects of vitamin C combined with antisense cyclin DI on B[a]P-induced changes of cell cycle were similar to those of vitamin C alone or antisense cyclin DI alone. Conclusions B[a]P progressed HELF cells from G1 to S phase via intracellular signaling pathway of cyclin D I/E2F. Vitamin C may modulate this signaling pathway to protect cells from injury caused by B[a]P.

Dose-dependent effects of lead on cell-cycle arrest, DNA damage, and cyclin D1 expression in primary cultured rat hippocampal neurons

BACKGROUND： Previous studies have suggested that the hippocampus is one of the neurotoxic target sites for lead. However, the molecular mechanisms of action, including the effect of lead on cell-cycle arrest, remain poorly understood. OBJECTIVE： To investigate the effects of different lead concentrations on cell-cycle arrest, DNA damage, and cyclin D1 expression in primary cultured rat hippocampal neurons. DESIGN, TIME AND SETTING： A randomized, controlled, in vitro experiment was performed at the China Medical University between July 2008 and May 2009. MATERIALS： Antibodies specific to cyclin D1 and actin were synthesized and purified by Santa Cruz Biotechnology, USA. FACStar flow cytometer was purchased from Becton Dickinson, San Jose, California, USA. METHODS： Wistar rat hippocampal neurons were primary cultured for 7 days. Neurons in the control group were treated with 0.01 mol/L phosphate buffered saline. Neurons in the 0.2, 1.0, and 10 umol/L lead acetate groups were subjected to 0.2, 1.0, and 10 umol/L lead acetate. Subsequently hippocampal neurons in each group were cultured for 24 hours. MAIN OUTCOME MEASURES： The effects of lead on cell cycle were measured by flow cytometry, DNA damage was measured using the comet assay, and cyclin D1 expression was measured using Western blot analysis. RESULTS： Treatment of hippocampal neurons with 0.2 umol/L lead acetate did not significantly alter cell cycle phase distribution, i.e., sub-G1, S, G0/G1, G2/M, whereas treatment with 1.0 and 10 umol/L lead acetate significantly increased the percentage of S and sub-G1 phase cells （P 〈 0.05）. Olive tail moment in all lead-treated groups and the percentage of DNA in the tail in 1.0 umol/L and 10 umol/L lead acetate groups were significantly greater compared with the control group （P 〈 0.05）. In addition, the percentage of tail DNA was greater in the 0.2 umol/L lead acetate group compared with the control group （P 〉 0.05）. Following incubation with 0.2, 1.0, and 10 umol/L lead acetate for 24 hours, cyclin D1 expression gradually decreased with exposure to increasing lead acetate concentrations （1.0-10 umol/L）. CONCLUSION： Lead exposure to primary cultured rat hippocampal neurons resulted in dose-dependently disturbed cellular homeostasis, including DNA damage, reduced cyclin D1 expression, and stagnation of cell-cycle progression.

Curcumin down-regulates PCNA,cyclin D1 and Bcl-X_L expression in human keratinocyte cell lines

目的将在在 keratinocyte 房间线调整增长，房间周期分发， apoptosis 和相关机制上评估 curcumin 的效果。人使人的 keratinocyte 线(HaCaT 房间) 不朽的方法与 curcumin 的不同剂量被对待。房间生存能力上的 curcumin 的效果被 MTT 试金测量，并且房间周期分发和 apoptosis 由流动 cytometry 决定了。原子抗原(PCNA ) ， cyclin D1 和 Bcl-xL 从即时 PCR 分析和蛋白质层次的增殖的房间的 mRNA 表示变化被西方的弄污检测。在学习获得的结果数据证明 curcumin 能在一个时间依赖者和剂量依赖者举止在 HaCaT 房间在增长上引起显著地禁止的效果。在 G1/S 阶段和重要 apoptosis 的房间拘捕在为 24 h 与 curcumin 被对待以后被观察。与这些联合， PCNA， cyclin D1 和 Bcl-xL 的表示为一样的处理在 mRNA 和蛋白质层次两个都被减少。结论 Curcumin 能禁止增长，在 G1/S 阶段导致房间拘捕并且引起在 HaCaT 房间的 apoptosis。curcumin 导致的 PCNA， cyclin D1 和 Bcl-xL 的减少的表示在 vitro 贡献上述效果。

THE OVEREXPRESSION AND SIGNIFICANCE OF CYCLIN D1 AND P53 IN CERVICAL SQUAMOUS CELL CARCINOMAS

Objective To investigate the significance of overexpresson of cyclin D1 and P53 protein in cervical squamous cell carcinomas.Methods Fifty cases of invasive cervical squamous cell carcinomas and 10 cases of normal cervical squamous epithelia were investigated with immunihistochemical technique. Results The overexpression of cyclin D1 and P53 in invasive cervical carcinomas was 70% and 50%, respectively. There was no overexpression of them in the control group. The overexpression of cyclin D1 in grade Ⅱ and Ⅲ was much higher than that in gradeⅠ(P<0.05). The overexpresson of cyclin D1 in stage Ⅲ of cervical carcinoma was significantly higher than that in stage Ⅱ (P<0.05). The overexpression of P53 in grade Ⅱ and grade Ⅲ of cervical carcinoma was remarkably higher than that in grade Ⅰ (P<0.05).Conclusion The action point of both cyclin D1 and P53 may be at G1/S transition. The overexpression of them was associated with development and progression of cervical carcinoma probably in different mechanisms and different pathways.

Inhibition of Mahkota Dewa(Phaleria macrocarpa) bioactive fraction on proliferation of human retinoblastoma tumor cells Y-79 through suppression of mRNA level of cyclin E

Objective: To prove the molecular mechanisms of Mahkota Dewa(Phaleria macrocarpa) in suppressing proliferation of human retinoblastoma cells through suppression of cell cycle's gene-regulators expression.Methods: In this study, the molecular mechanism of anti-tumor effect of fractioned extract of Phaleria macrocarpa(DLBS1425) in human retinoblastoma cells Y-79 was investigated by measuring the tumor cells viability, the assessment of population profiles of tumor cells in the cell cycle, and the mRNA concentration of p16, p21, p53, cyclin D,cyclin E, and E2 F.Results: DLBS1425 showed an inhibition effects towards proliferation of Y-79 cell line.Inhibition of proliferation was shown by suppression of cell cycle progression.DLBS1425 downregulated cyclin E, a G1 phase regulator gene of cell cycle, in dosedependent manner without affecting p53–p21 pathway.In the other word, DLBS1425 inhibits cell proliferation through suppression of cyclin E independently towards conventional proliferation pathway.Conclusions: Our results suggest that DLBS1425 is a potential anticancer agent which targets genes involved in cell proliferation in human retinoblastoma cells which make it pharmacologically ideal for the prevention and/or treatment of retinoblastoma cancer.

Cyclin D1、Ki-67、CK19联合评估甲状腺乳头状癌的效能及与颈部淋巴结转移的相关性分析

目的:探讨细胞周期蛋白D1(Cyclin D1)、细胞增殖抗原-67(Ki-67)、细胞角蛋白19(CK19)联合评估甲状腺乳头状癌(PTC)的效能,分析三者与颈部淋巴结转移的相关性。方法:选取我院2017年9月至2021年9月收治的91例PTC患者为PTC组,取同期收治的结节性甲状腺肿患者52例为对照组。两组均通过手术获取病变组织,检测Cyclin D1、Ki-67、CK19表达情况,分析三者对PTC的诊断价值。根据PTC患者有无颈部淋巴结转移,分成转移组、非转移组,比较两组病理组织中Cyclin D1、Ki-67、CK19表达情况,经多元Logistic回归模型分析三者与PTC患者颈部淋巴结转移的关系。结果:PTC组Cyclin D1、CK19阳性表达率分别为83.52%、85.71%,高于对照组的17.31%、17.31%(P<0.05);PTC组Ki-67>1%占比为93.41%,高于对照组的13.46%(P<0.05)。Cyclin D1、Ki-67、CK19单独诊断PTC的敏感度分别为83.52%、93.41%、85.71%,特异度分别为82.69%、86.54%、82.69%,三者联合诊断的敏感度、特异度分别为91.21%、96.15%。转移组的Cyclin D1、CK19阳性表达率分别为97.06%、97.06%,高于非转移组的75.44%、78.95%(P<0.05)。多元Logistic回归分析显示,Cyclin D1阳性表达会增加PTC患者颈部淋巴结转移风险(OR:3.446,95%CI:1.041~11.407,P<0.05)。结论:PTC患者的Cyclin D1、Ki-67、CK19阳性表达率较高,三者联合对PTC有较好的诊断效能,其中Cyclin D1阳性表达与颈部淋巴结转移相关。

cyclin D1表达模式深度学习识别和预测模型对HPV阴性口腔鳞状细胞癌和口咽鳞状细胞癌患者预后的预测作用

目的:通过评估cyclin D1的表达状态与HPV阴性口腔鳞状细胞癌(oral squamous cell carcinoma, OSCC)及口咽鳞状细胞癌(oropharyngeal squamous cell carcinoma, OPSCC)患者预后的关系,建立基于cyclin D1表达模式的图像识别评分和生存预测模型。方法:回顾性分析610例HPV阴性OSCC和OPSCC患者的临床病理资料。通过比较cyclin D1在不同评价方式以及联合p16/p53表达等多种因素评价体系下患者总体生存(OS)及无进展生存(PFS)的差异,检测cyclin D1对OSCC、OPSCC患者预后的预测作用。利用YOLOv5图像识别算法建立cyclinD1表达模式评分模型,并在此基础上用DeepHit和DeepSurv算法分别建立预后模型。结果:cyclin D1在OSCC和OPSCC癌巢中存在三级表达模式。该表达模式与OSCC(P<0.0001)、OPSCC(P<0.05)患者预后显著相关,且优于cyclin D1表达水平与患者预后的相关性。cyclin D1表达模式在OSCC(P<0.0001)及OPSCC(P<0.05)中均为独立的预后风险因素。基于cyclin D1表达模式的评分模型在测试集中的准确率达(78.48±4.31)%。在OS预测中,DeepHit算法建立的模型在测试集中的C-index为0.709±0.019,DeepSurv算法建立模型测试集中C-index为0.715±0.029。结论:基于深度学习的cyclin D1表达模式评分模型联合生存预测模型对HPV阴性OSCC和OPSCC总体生存预后具有较好的预测作用。

Significance of cyclin E, p16ink4a and ki67 Overexpression in Cervical Exfoliated-cell Specimens for Primary Screening of HPV-related Cervical Carcinoma

The aim of this study is to investigate cyclin E, pl6ink4a and ki67 as possible diagnostic biomarkers for cervical preneoplasia using cervical exfoliated-cell specimens, and evaluate the significance for screening patients at high risk of de- veloping cervical carcinoma. The expression of cyclin E, p16ink4a and ki67 was examinated in 78 cervical exfoliated epithe- lial specimens diagnosed as atypical squamous cells of undetermined significance (ASCIIS) (12 cases), cervical intraepithe- lial neoplasia (CIN) of type 1 (17 cases), CIN2-3(38 cases) and invasive carcinoma (11 cases) using immunohistochemical analysis, and simultaneously, the DNA status of human papillomavirus (HPV) type 16/18 was detected by polymerase chain reaction (PCR) using type specific primers, cyclin E, pl6ink4a and ki67 were all overexpressed in CINs and invasive carci- noma, compared with little expression in ASCUS ( P < 0.005). Overexpression of cyclin E was observed in CIN1 (94.1 % , x2 = 21.16, P < 0.01), and pl6ink4a and ki67 were overexpressed in invasive carcinoma (100% and 90.9% respective- ly) . The degree of pl6ink4a and ki67 expression correlated well with that of epithelial lesions ( P < 0.005). HPV16/18 infec- tion was assessed in CINs and invasive carcinoma samples, and revealed a significant relationship with the degree of cervical epithelial lession. The expression level of pl6ink4a and ki67 seemed more closely associated with HPV16 infection than that of cyclin E ( r s = 1.0 vs rs = 0 . 4 ) . Only 1 case in CIN, and 4 cases in CIN2-3 of HPV18 positive samples were detected. Therefore no statistical significance was found by statistical analysis. Overexpression of cyclin E, p16ink4a and ki67 in CINs and invasive carcinoma cells demonstrates the potential use of cyclin E, p16ink4a and ki67 as diagnostic biomarkers for HPV-related cervical neoplastic lesions. In addition, this technique can be used for screening patients at high risk of devel- oping cervical carcinoma.

miR-567通过调控CDK8在NSCLC增殖、迁移和细胞周期中的作用及其临床相关性研究

目的探讨微小RNA(miR)-567通过调控周期蛋白依赖性激酶8(CDK8)在非小细胞肺癌(NSCLC)增殖、迁移和细胞周期中的作用及其临床相关性研究。方法收集40例NSCLC患者的肿瘤组织和临近癌旁组织,采用实时荧光定量PCR(qRT-PCR)检测miR-567和CDK8的表达。将miR-NC mimic、miR-567 mimic、oe-NC和oe-CDK8转染至A549和H1975细胞中,使用qRT-PCR检测miR-567和CDK8的表达,CCK-8法检测细胞增殖水平,Transwell法检测细胞迁移水平,流式细胞术检测细胞周期变化。通过荧光素酶报告基因实验检测miR-567与CDK8的靶向性。结果在NSCLC患者的肿瘤组织中,miR-567表达降低,而CDK8表达升高,二者呈负相关(P<0.05)。在A549和H1975细胞中,miR-567 mimic组相较于miR-NC mimic组,miR-567表达升高,CDK8表达降低,细胞增殖和迁移水平降低,细胞G1期比例升高,S期比例降低;miR-567 mimic组在正常型CDK8中,荧光强度低于miR-NC mimic组;miR-567 mimic+oe-CDK8组相较于miR-567 mimic+oe-NC组,CDK8表达升高,细胞增殖和迁移水平升高,细胞G1期比例降低,S期比例升高。结论miR-567通过靶向抑制CDK8表达,控制肿瘤细胞在S期阻滞,从而抑制NSCLC的增殖和迁移能力。

Effects of estradiol on cell cycle and cyclin proteins of vascular smooth muscle cells in rats

To study the effects of 17β-estradiol(E2) on the growth of cultured rat vascular smooth muscle cells (VSMC).Methods The cell cycle and the expressions of Cyclin D1 and CDK4 proteins were examined by flow cytometry in VSMC cultured in different concentrations (0～100 nmol/L) of 17β-estradiol with or without serum.Results Under serum-stimulating conditions,17β-estradiol(1,10,100 nmol/L) promoted VSMC proliferation by accelerating their cell cycle progression from G1 to S phases,and the cell rates at S were (31.89±9.14)%(35.90±4.59)% and (30.77±1.20)% respectively,significantly higher than the corresponding values of control cells (21.63±1.80)%.This was accompanied by the significantly increased expression of Cyclin D1 and CDK4 proteins.In the cultures without serum,however,high concentrations (10,100 nmol/L) of E2 induced a cell cycle arrest at G1 phase,which was characterizsed by decreased cell rates at S phase [(9.93±1.43)% and (8.76±1.80)% respectively,P<0.05] as compared with the corresponding control values and a down-regulation of expressions of Cyclin D1 and CDK4 proteins.Conclusion E2 can either promote or inhibit VSMC proliferation depending upon the presence or absence of serum mitogens.The underlying mechanism may be associated with the hormone’s action on the expression of Cyclin D1 and CDK4 which act as the G1 phase regulators.4 refs.