1-Chloromethyl-6,7-dimethoxy-3,4-dihydro-1H-isoquinoline-2-sulfonic acid amide, a derivative of tetrahydroisoquinoline, induces granulocytic differentiation of the human leukemic HL-60 cells via G0/G1 phase arrest

Tetrahydroisoquinolines are known to have various biological effects, including antitumor activity. This study investigated the effect of 1-chloromethyl-6, 7-dimethoxy-3, 4-dihydro-1H-isoquinoline-2-sulfonic acid amide (CDST), a newly synthesized anticancer agent, on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells were determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide staining, western blot analysis and immunoprecipitation, respectively. CDST induced the differentiation of HL-60, as shown by increased expression of differentiation surface antigen CD11b (but no significant change in CD14 expression) and increased NBT-reducing functional activity. DNA flow cytometry analysis indicated that CDST markedly induced a G0/G1 phase arrest of HL-60 cells. Subsequently, we examined the expre-ssion of G0/G1 phase cell cycle-related proteins, including cyclin-dependent kinases (CDKs), cyclins and cyclin dependent kinase inhibitors (CKIs), during the differentiation of HL-60. The levels of CDK2, CDK6, cyclin E and cyclin A were decreased, whereas steady-state levels of CDK4 and cyclin D1 were unaffected. The expression of the p27Kip1 was markedly increased by CDST, but not p21WAF1/Cip1. Moreover, CDST markedly enhanced the binding of p27Kip1 with CDK2 and CDK6, resulting in the reduced activity of both kinases. Taken together, these results demonstrate that CDST is capable of inducing cellular differentiation and growth inhibition through p27Kip1 protein-related G0/G1 phase arrest in HL-60 cells.

Nanosize aminated fullerene for autophagic flux activation and G0/G1 phase arrest in cancer cells via post-transcriptional regulation

Functional fullerene derivatives exhibit special inhibitory effects on tumor progress and metastasis via diverse tumor microenvironment regulations,while the elusive molecular mechanisms hinder their clinical transformation.Herein,it is initially revealed that nanosize aminated fullerene(C_(70)-EDA)can activate autophagic flux,induce G0/G1 cell cycle arrest to abrogate cancer cell proliferation,and significantly inhibit tumor growth in vivo.Mechanismly,C_(70)-EDA promotes the expression of cathepsin D involved in autophagic activation via post-transcriptional regulation,attributing to the interaction with a panel of RNA binding proteins.The accumulation of cathepsin D induces the autophagic degradation of cyclin D1,which arouses G0/G1 phase arrest.This work unveils the fantastic anti-tumor activity of aminated fullerene,elucidates the molecular mechanism,and provides a new strategy for the antineoplastic drug development on functional fullerenes.

Krill oil ameliorates benign prostatic hyperplasia by regulating G1-phase cell cycle arrest and altering signaling pathways and benign prostatic hyperplasia-associated markers

Krill oil(KO)exhibits various biological activities,such as anti-inflammatory and antitumor effects.However,the inhibitory effects of benign prostatic hyperplasia(BPH)in vitro and in vivo have not yet been studied.This study investigated the anti-BPH effects of KO extracted by an enzymatic hydrolysis method.KO treatment inhibited the proliferation of WMPY-1 and BPH-1 cells by induction of G0/G1 phase arrest through the modulation of positive and negative regulators in both prostate cell types.KO treatment stimulated phosphorylation of c-Jun N-terminal kinase(JNK)and p38 signaling.In addition,KO changed the expression of BPH-related markers(5α-reductase,androgen receptor,FGF,Bcl-2,and Bax)and the activity of the proliferation-mediated NF-κB binding motif.KO-induced levels of proliferation-mediated molecules of prostate cells were attenuated in the presence of siRNA-specific p-38(si-p38)and JNK(si-JNK).Furthermore,the administration of KO alleviated prostate size and weight and the cell layer thickness of prostate glands in a testosterone enanthate-induced BPH rat model.KO treatment altered the level of dihydrotestosterone in serum and the expression levels of BPH-related markers in prostate tissues.Finally,KO-mediated inhibition of prostatic growth was validated by histological analysis.These results suggest that KO has an inhibitory effect on BPH in prostate cells in vitro and in vivo.Thus,KO might be a potential prophylactic or therapeutic agent for patients with BPH.

贝母素乙通过诱导G0/G1期阻滞抑制结肠癌细胞的增殖

目的:探讨贝母素乙对结肠癌HCT116细胞增殖的抑制作用及其分子机制。方法:利用不同浓度的贝母素乙处理人结肠癌细胞HCT116和正常结肠上皮细胞CCD841 CON,通过CCK-8法和结晶紫染色法检测贝母素乙对HCT116和CCD841 CON细胞增殖活力的影响,流式细胞术和WB法检测贝母素乙对HCT116细胞周期及其细胞周期相关蛋白表达的影响。构建HCT116移植瘤裸鼠模型和AOM/DSS结肠癌小鼠模型,观察贝母素乙对小鼠模型肿瘤生长和OS的影响,免疫组化法和WB法检测对移植瘤或肿瘤组织中细胞周期相关蛋白CDK4、CDK6和cyclin D1表达的影响。结果:贝母素乙可显著抑制结肠癌HCT116细胞的增殖能力(P<0.01),诱导HCT116细胞周期G0/G1期阻滞(P<0.01),降低CDK4、CDK6和cyclin D1的蛋白表达水平(均P<0.01)。荷瘤小鼠实验结果显示,贝母素乙(0.75 mg/kg)显著抑制HCT116细胞移植瘤的生长并延长荷瘤裸鼠的OS(P<0.05或P<0.01),降低AOM/DSS模型小鼠的体质量、延长OS、减少癌变肠组织的肿瘤个数和肿瘤体积,下调肿瘤组织中CDK4、CDK6和cyclin D1的蛋白表达(P<0.01或P<0.05)。结论:贝母素乙通过下调CDK4、CDK6和cyclin D1的表达水平,引起细胞周期G0/G1期阻滞,从而抑制结肠癌HCT116细胞的增殖。

Chaga mushroom (Inonotus obliquus) induces G0/G1 arrest and apoptosis in human hepatoma HepG2 cells

AIM: To investigate the anti-proliferative and apoptotic effects of Chaga mushroom (Inonotus obliquus) water extract on human hepatoma cell lines,HepG2 and Hep3B cells. METHODS: The cytotoxicity of Chaga extract was screened by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Morphological observation,flow cytometry analysis,Western blot were employed to elucidate the cytotoxic mechanism of Chaga extract. RESULTS: HepG2 cells were more sensitive to Chaga extract than Hep3B cells,as demonstrated by markedly reduced cell viability. Chaga extract inhibited the cell growth in a dose-dependent manner,which was accompanied with G0/G1-phase arrest and apoptotic cell death. In addition,G0/G1 arrest in the cell cycle was closely associated with down-regulation of p53,pRb,p27,cyclins D1,D2,E,cyclin-dependent kinase (Cdk) 2,Cdk4,and Cdk6 expression. CONCLUSION: Chaga mushroom may provide a new therapeutic option,as a potential anticancer agent,in the treatment of hepatoma.

Intrinsic apoptotic pathway and G2/M cell cycle arrest involved in tubeimoside I-induced EC109 cell death

Objective： Squamous esophageal carcinoma is highly prevalent in developing countries, especially in China. Tu Bei Mu （TBM）, a traditional folk medicine, has been used to treat esophageal squamous cell carcinoma （ESCC） for a long term. tubeimoside I （TBMS1） is the main component of TBM, exhibiting great anticancer potential. In this study, we investigated the mechanism of TBMS1 cytotoxic effect on EC109 cells. Methods： Comparative nuclear proteomic approach was applied in the current study and we identified several altered protein spots. Further biochemical studies were carried out to detect the mitochondrial membrane potential, cell cycle and corresponding proteins＇ expression and location. Results： Subcellular proteomic study in the nucleus from EC109 cells revealed that altered proteins were associated with mitochondrial function and cell proliferation. Further biochemical studies showed that TBMSl-induced molecular events were related to mitochondria-induced intrinsic apoptosis and P21-cyclin B 1/cdc2 complex-related G2/M cell cycle arrest. Conclusions： Considering the conventional application of TBM in esophageal cancer, TBMS1 therefore may have a great potential as a chemotherapeutic drug candidate for ESCC.

Ethanol extract of Kalopanax septemlobus leaf inhibits HepG2 human hepatocellular carcinoma cell proliferation via inducing cell cycle arrest at G_1 phase

Objective:To investigate the effects of an ethanol extract of Kalopanax septemlobus(Thunb.)Koidz.leaf(EEKS) on cell proliferation in human hepatocellular carcinoma cells and its mechanisms of action.Methods:Cells were treated with EEKS and subsequently analyzed for cell proliferation and flow cytometry analysis.Expressions of cell cycle regulators were determined by reverse transcriptase polymerase chain reaction analysis and Western blotting,and activation of eyclin-associaled kinases studied using kinase assays.Results:The EEKS suppressed cell proliferation in both HepG2 and Hep3 B cells,but showed a more sensitive anli-proliferative activity in HepG2 cells.Flow cytometry analysis revealed an association between the growth inhibitory effect of EEKS and with G_1 phase cell cycle arrest in HepG2 cells,along with the dephosphorylation of retinoblastoma protein(pRB) and enhanced binding of pRB with the E2 F transcription factor family proteins.Treatment with EEKS also increased the expression of cyclin-dependent kinase(CDK) inhibitors,such as p21WAF1/CIP1 and p27KIP1.without any noticeable changes in G_1 cyclins and CDKs(except for a slight decrease in CDK4).Treatment of HepG2 cells with EEKS also increased the binding of p21 and p27 with CDK4 and CDK6.which was paralleled by a marked decrease in the cyclin D- and cyclin E-associated kinase activities.Conclusions:Overall,our findings suggest that EEKS may be an effective treatment for liver cancer through suppression of cancer cell proliferation via G_1,cell cycle arrest Further studies arc required to identify the active compounds in EEKS.

阳性表达表皮钙粘蛋白增加G0/G1期人乳腺癌细胞比例及其分子机制

表皮钙粘蛋白 (E cadherin)阴性的乳腺癌细胞株MDA MB 2 3 1和MDA MB 43 5转染野生型表皮钙粘蛋白基因 ,通过流式细胞仪测量细胞周期发现表皮钙粘蛋白阳性细胞生长变慢 ,更多细胞停滞在G0 /G1期 ,蛋白质印迹证实由G0 /G1期进入S期的重要调控分子细胞周期蛋白 D1(cyclinD1)下降了 ,并发现表皮钙粘蛋白还能降低直接激活细胞周期蛋白 D1基因转录的 β 连环蛋白的蛋白质浓度 .蛋白激酶B (PKB)能通过抑制糖原合成激酶 3 β(GSK 3 β)的活性来抑制β 连环蛋白降解 ,并在乳腺癌高转移细胞株中普遍过表达 ,其表达同样受到了表皮钙粘蛋白的抑制 .并且在表皮钙粘蛋白阳性细胞中 ,作为PKB上游信号分子并能激活PKB的粘着斑激酶 (FAK)和整联蛋白相关激酶 (ILK)蛋白量也发生下降 ,能抑制PKB激活的PTEN蛋白量却增加了 .结果显示 ,表皮钙粘蛋白能通过降低乳腺癌细胞中的PKB蛋白浓度 ,并通过上游信号分子抑制PKB的激活 ,进而降低PKB对β 连环蛋白降解的抑制作用 ,导致 β 连环蛋白直接调控的靶基因细胞周期蛋白D1的表达量下降 ,引起更多的细胞停止在G0

Butyrate inhibits the bovine rumen epithelial cell proliferation via downregulation of positive regulators at G0/G1 phase checkpoint

Short-chain fatty acids(SCFAs)butyrate promote the postnatal rumen epithelial development and maturation in ruminants.However,molecular mechanisms of effects of butyrate on the bovine rumen epithelial cells(BRECs)proliferation remain elusive.Therefore,purpose of this study was to investigate the effects of butyrate on the expression of genes and proteins at G0/G1 and S phase of BRECs cycle.Our results showed that BRECs treated with butyrate inhibited(P<0.05)the proliferation of BRECs,relatively to control.Flow cytometric assays revealed that butyrate triggers the BRECs cycle arrest at the G0/G1 phase.qRT-PCR analyses of mRNA level of genes involved in the G0/G1 phase of cell cycle showed that butyrate significantly upregulated(P<0.001)the expression of mRNA encoding p21^(Cip1)compared with control group,but it decreased(P<0.05)the mRNA levels of cyclin D1 and CDK4 genes at G0/G1 phase checkpoint compared with control.Moreover,Western blot also revealed that butyrate downregulated the expression of cyclin D3,CDK6,p-Rb,and E2F1 proteins involved in the modulation of G0/G1 phase of cell cycle.In conclusion,our results demonstrated that butyrate inhibits the proliferation of BRECs via downregulation of positive regulators at G0/G1 phase checkpoint.

对数生长与G_0期人包皮成纤维细胞Stat1对INF-α刺激反应的差异

Stat1 (信号转导与转录活化子 1 )的激活与刺激因子和细胞组织类型有关 ,它的活化受体有组织分布特异性 .本研究比较 G0 期和对数生长周期两个不同状态体外培养的人包皮成纤维细胞受到干扰素 ( INF-α)刺激后 Stat1基因 m RNA、蛋白质表达以及蛋白质合成效率的差异 .结果表明 :成对数生长的细胞 Stat1的m RNA表达量较对照组增加达 1 0倍左右 ,蛋白质表达增加 3～ 4倍左右 ;G0 期细胞 Stat1的 m RNA表达量较对照组增加约 3～ 4倍 ,蛋白质表达增加约 3～ 4倍 ;对数生长细胞 Stat1蛋白质合成效率明显降低 ,在翻译水平可能有调控 .以上结果提示对数生长和 Go 细胞对

M/G/1排队系统算子生成正压缩C_0-半群的不可约性

M/G/1排队系统已有大量文献研究.通过增补变量法,该系统可由一组积分微分方程描述,并且系统算子在L^1空间中生成正的压缩C_0-半群.文中将进一步讨论该半群的性质,证明该半群是不可约的.

N-cadherin阳性白血病KG1a细胞系在G_0期抵抗VP16杀伤的作用

抗药性是白血病干细胞的重要特征,为探索N-cadherin阳性的白血病细胞耐受化疗药物VP16杀伤作用的机制,本研究以白血病细胞系KG1a为研究模型,利用流式细胞术测定N-cadherin阳性和N-cadherin阴性细胞在G0期比例的差异,利用G-CSF诱导KG1a细胞进入细胞周期,观察G0期细胞比例的变化,并测定诱导后KG1a细胞对VP16的敏感性;再利用EGTA抑制N-cadherin介导的细胞间黏附后,观察KG1a细胞耐药性的变化。结果显示,N-cadherin阳性的KG1a细胞G0期比例高于N-cadherin阴性的细胞;诱导KG1a细胞进入细胞周期后G0期细胞比例明显下降,KG1a细胞对VP16的敏感性显著升高;利用EGTA处理KG1a细胞24小时抑制N-cadherin的作用后,KG1a细胞在G0期比例降低,KG1a细胞对VP16的药物敏感性显著升高。结论:N-cadherin通过介导白血病细胞之间的黏附作用,使白血病细胞处于G0期的静息状态,从而耐受VP16的杀伤作用。

奥曲肽通过G_0/G_1期阻滞抑制胆管癌细胞的增殖

为观察四种胆管癌细胞系 (RBE、NEC、QBC939、SSP 2 5 )是否能够表达生长抑素受体 (SSTR) ,以及八肽生长抑素类似物奥曲肽 (OCT)对胆管癌细胞的抑制作用 ,采用逆转录 聚合酶链反应方法检测四种细胞系 2 ,3型SSTR的表达 ;四氮唑蓝法检测不同浓度OCT (10、 1、 0 1、 0 0 1和 0 0 0 1mg L)在体外对胆管癌细胞增殖的影响。并应用碘化丙啶 (PI)及AnnexinV PI染色后于流式细胞仪检测经OCT作用后胆管癌细胞的细胞周期和凋亡情况。结果显示四种胆管癌细胞系均可以表达SSTR2 mRNA ,但未检测到SSTR3mRNA的表达。体外 10、 1和0 1mg LOCT明显抑制四种胆管癌细胞的增殖 (与各自对照组相比 ,P <0 0 5 ) ;1mg LOCT作用 4 8h后流式细胞分析表现为G0 G1 期细胞增加 ,S期和G2 M细胞减少 (与各自对照组相比 ,P <0 0 1) ,但没有明显凋亡发生。从而证明OCT抑制胆管癌细胞增殖的机制主要与引起细胞周期阻滞有关 ,而不是促进凋亡。对于表达生长抑素受体的胆管癌 。

具有可选服务及无等待空间的M/G/1排队系统算子生成的正压缩C_0-半群的不可约性

具有可选服务及无等待空间的M/G/1排队系统算子生成正压缩C0—半群,本文进一步讨论了该半群的不可约性。

桑黄水提物诱导黑色素瘤细胞系B16-F10 G_(0)/G_(1)期阻滞的研究

通过噻唑蓝(MTT)比色法、流式细胞分析、转录组测序和蛋白印迹等手段,探究桑黄水提物(SH)对黑色素瘤细胞系B16-F10的抑制作用及分子作用机制。MTT实验结果显示,SH能够抑制B16-F10细胞系增殖,且呈剂量依赖性。流式细胞分析发现,SH能够诱导细胞G_(0)/G_(1)期阻滞,但对细胞凋亡无明显影响。RNA-seq、qRT-PCR和Western blot分析发现,SH主要通过上调p53信号通路中p21的表达,进而抑制细胞周期通路中CyclinD3、CDK2和CDK6的表达来影响细胞周期阻滞。研究结果证明SH对黑色素瘤细胞增殖具有显著的抑制作用,p21-CyclinD3-CDKs信号通路介导的G_(0)/G_(1)期阻滞是其作用机制之一。

催化剂CS-1-G用于生产1102K型聚丙烯树脂的工业试验

介绍了15万t/a Novolen气相法聚丙烯(PP)装置采用国产催化剂CS-1-G生产1102 K型PP树脂的工业试验情况,并与装置原用进口催化剂进行了对比。结果表明,在原料、助剂性质及生产工艺条件基本接近的情况下,催化剂CS-1-G的活性达到56.86 kg/g,比进口催化剂高出7.75%;装置运行稳定,所产PP产品的细粉含量少,其主要性能指标达到进口催化剂生产产品的水平。

胆碱拮抗剂R2HBJJ诱导非小细胞肺癌细胞G0／G1期阻滞并抑制细胞增殖

目的：评价胆碱拮抗剂R2HBJJ对非小细胞肺癌细胞生长的影响，探讨其作用机制。方法：SRB法测定细胞活力，RT—PCR检测受体表达，

Circadian variation in expression of G_1 phase cyclins D_1 and E and cyclin-dependent kinase inhibitors p16 and p21 in human bowel mucosa

AIM： To evaluate whether the cellular proliferation rate in the large bowel epithelial cells is characterized by circadian rhythm. METHODS： Between January 2003 and December 2004, twenty patients who were diagnosed as suffering from primary, resectable, non-metastatic adenocarcinoma of the lower rectum, infiltrating the sphincter mechanism, underwent abdominoperineal resection, total mesorectal excision and permanent left iliac colostomy. In formalinfixed and paraffin-embedded biopsy specimens obtained from the colostomy mucosa every six hours （00：00, 06：00, 12：00, 18：00 and 24：00）, we studied the expression of G1 phase cyclins （D1 and E） as well as the expression of the G1 phase cyclin-dependent kinase （CDK） inhibitors p16 and p21 as indicators of cell cycle progres- sion in colonic epithelial cells using immunohistochemical methods. RESULTS： The expression of both cyclins showed a similar circadian fashion obtaining their lowest and highest values at 00：00 and 18：00, respectively （P〈 0.001）. A circadian rhythm in the expression of CDK inhibitor proteins p16 and p21 was also observed, with the lowest levels obtained at 12：00 and 18：00 （P〈0.001）, respectively. When the complexes cyclins D1-p21 and E-p21 were examined, the expression of the cyclins was adversely correlated to the p21 expression throughout the day. When the complexes the cyclins D1-p16 and E-p16 were examined, high levels of p16 expression were correlated to low levels of cyclin expression at 00：00, 06：00 and 24：00. Meanwhile, the highest expression levels of both cyclins were correlated to high levels of p16 expression at 18：00. CONCLUSION： Colonic epithelial cells seem to enter the G1 phase of the cell cycle during afternoon （between 12：00 and 18：00） with the highest rates obtained at 18：00. From a clinical point of view, the present results suggest that G1-phase specific anticancer therapies in afternoon might maximize their anti-tumor effect while minimizing toxicity.

mTORC1控制静息状态干细胞从G_0到G_(Alert)的适应性转变

许多成体干细胞的一个独特属性是其能以一个非循环的静息状态存在。在需要动用干细胞维持组织稳态或修复前,静息状态能一直维持干细胞的功能。然而,静息状态下的干细胞的缺陷可导致组织功能障碍。

大容量NAND FLASH K9T1G08U0M在汽车音响中的应用

随着汽车音响的不断发展,多媒体设备的使用随之增多,在汽车音响中加入大容量存储芯片是很必要的。以三星公司的K9T1G08U0M闪存为例,介绍了闪存的存储结构、功能特点、I/O口线的特点、专用的命令指令和状态寄存器。进一步介绍了在汽车音响设备中该芯片通过I/O口线通信来传递数据进行多媒体资料存储的方法。同时也给出了读写FLASH的程序实现流程方法。目前该方案已应用于车载音响设备。