Changes of the Microtubule Arrays During Mitosis in Prothallus Cells of Dryopteris crassirhizoma

Microtubule arrays in prothalli large-vacuolated and meristematic dividing cells of the fern Dryopteris crassirhizoma Nakai were studied using Steedman's wax, indirect immunofluorescence labelling and confocal laser scanning microscopy. Results showed that the use of high paraformaldehyde concentration (8%) allowed good fixation of prothallus cells, which are characterized by numerous (meristematic cells) and big (large-vacuolated cells) vacuoles. Results also plead for the efficiency of Steedman's wax embedding method in: (1) avoiding excessive use of enzyme for digesting cell wall in the process of the microtubule cytoskeleton labelling, (2) minimizing the autofluorescence effect in cells through utilization of alcohol in sample dehydration, and (3) permitting a clear visualization of microtubule patterns during the cell mitosis. Steedman's wax, coupled with immunofluorescence labelling and confocal laser scanning microscopy techniques, allows a good investigation of cell division process in plants by using simple multicellular organisms such as fern prothalli.

Inhibitory effect of Polo-like kinase 1 depletion on mitosis and apoptosis of gastric cancer cells

AIM： Polo-like kinase 1 （PLK1） serine/threonine kinase plays a vital role in multiple phases of mitosis in gastric cancer cells. To investigate the effect of PLK1 depletion on mitosis and apoptosis of gastric cancer cells. METHODS： PLK1 expression was blocked by small RNA interference（siRNA）. The expression levels of PLK1, cdc2, cyclin B and caspase 3 were detected by Western blotting. Then, PLK1 depletion, cdc2 activity, cell proliferation, cell cycle phase distribution, mitotic spindle structure, and the rate of apoptosis of the PLK1 knockdown cells were observed. RESULTS： PLK1 gene knockdown was associated with increased cyclin B expression, increased cdc2 activity （but not with the expression levels）, accumulation of gastric cancer cells at G2/M, improper mitotic spindle formation, delayed chromosome separation and delayed or arrested cytokinesis. Moreover, PLK1 depletion in gastric cancer cells was associated with decreased proliferation, attenuated pro-caspase 3 levels and increased apoptosis. CONCLUSION： Blockage of to decreased mitosis or even PLK1 expression may lead apoptosis in gastric cancer cells, indicating that PLK1 may be a valuable therapeutic target for gastric cancer.

Mitosis and microtubule organizational changes in rice root-tip cells

The pattern of change of the microtubule cytoskele-ton of the root-tip cells of rice during mitosis was studied using immunofluorescence technic and confocal laser scanning microscopy. All the major stages of cell division including preprophase, prophase, metaphase, anaphase and telophase were observed. The most significant finding was that in the preprophase cells microtubules radiating from the nuclear surface to the cortex were frequently seen. During development these microtubules became closely associated with the preprophase band and prophase spindle indicating that the microtubules radiating from the nuclear surface, the preprophase band and the prophase spindle were structurally and functionally closely related to each other. Granule-like anchorage sites for the radiating microtubules at the nuclear surface were often seen and the possibility that these granule-like anchorage sites might represent the microtubule organizing centres was discussed.

Induced Differentiation of Epithelioid Carcinoma Cell Lines: Evidence for Tumor Cell Quantal Mitosis

The effects of growth factors and calcium concentrations present in different culture media on induction of terminal differentiation were investigated for four different epidermoid carcinoma cell lines, Hela, KB, A431, and SCC-25, and their responses determined relative to those elicited by normal human keratinocytes subjected to these culture conditions. Differentiation status was determined cyto-chemically by a validated keratin protein staining method, and by autoradiographic analyses. Growth and differentiation promoting factors that influenced the direction of integrated control of growth and differentiation in normal human keratinocytes were found to be effective for some cell lines but not others. The factors examined were 1) high density arrest in serum-free and serum-containing media, 2) media shifts from high density culture in serum-containing media to low density growth factor-depleted or supplemented serum-free medium, and 3) the concentration of calcium in the media. The extent and degree of differentiation achieved varied among different cell lines depend on the presence or absence of serum, EGF and insulin protein growth factors. Certain growth media appear to sponsor keratin protein, cyto-chemically-detected differentiation, and evidence of quantal mitotic division in low density HeLa cell and SCC25 cell cultures. Epidermoid carcinoma cell lines retain limited capacity to commit to early stages of cell differentiation.

Near-Dead Cells to Special Tetraploidy to First Cells to Cancer Diagnostic Morphology: Unlikely Therapy-Gain from For-Profit Industrial Goliath

The objective in this experimental article is to gain evidential proof of near-dead cells, (sick-cells in relapse tumor) responding with recovery growth from special 4n, multi-chromatid chromosomes. Note, near-dead normal human cells with such converted chromosome structure gave rise to proliferative, fitness-gained, diploid first cells, which further gave rise to three different cell shape changed, recovery growth patterns. Previously, two cell shape changes had been recovered from same type normal human cells, transiently exposed to amino acid glutamine deficient growth medium with recovery growths also associated with presence of the special 4n cells. The 4n cell-division had been concluded to be a meiotic-like two-step division system to the fitness-gained diploid cells in numerous experiments. The main characteristics of this division system, was firstly whole genomes without polar oriented bent centromeres moving apart followed by much rarer simple fission division to two or three diploid cells, selectable for first cell proliferation. In general these 4n cells showed metaphase type rosette figures moving apart not in the normal spindle associated mitotic shape with centromeres polar-pointing with sloping arms. This sequence of events induced by glutamine-deficiency, was earlier shown to cause DNA breakage in metabolic studies however, the near-death condition was only assumed from normal fibro-blastic cell-sheet shrinkage. This was rectified by an RNA virus (Coxakie-B3), which virology known is a highly cell killing virus (4+ CPE on their scale). This virus replicates only in replicating cells, which led to recovery growths with progressive phenotypic cell-shape changes (spindle, polygonal and roundness cells), each intervened by “total” cell destruction. These three different growth patterns had morphologies, indistinguishably from today’s cancer diagnostic morphologies. “Mitotic” analyses of beginning growths for the three phenotypes revealed the special rosette figure separations from special 4n and higher ploidy level cells, and also total absence of spindle type mitoses. Tumorigenesis-relevant was centromere-puffing with premature chromatid separation, and chromatin compaction, a mechanism, that was suggested to protect the genome from damage (text). We suggest that the multi-chromatid polyploid cells with their genome reductive division system, can be a tractable in vitro model system for therapy information, when repeated from a cell-killing agent, producing virus-free recovery growths. Will it be enacted upon? Not likely with profit-greedy industrial Goliath in the helm of cancer research. But, a not for profit cancer organization, could change this appalling situation.

VALUATION DE L'EXPRESSION DES RGULATEURS DU CYCLE CELLULAIRE EN G1/S DANS LE CARCINOME PIDERMO DE DE L'CSOPHAGE

Objective To study the expression of p16, cyclin D1, Rb and phosphorylated Rb(Ser795) in the esophageal squamous cell carcinoma. Methods Immunohistochemistry was performed and analysed on 34 cases of paraffin-embedded tissues. Results The expression of phosphorylated Rb(Ser795) was positively correlated to that of cyclin D1 (r= 0.401, P= 0.021) and inversely to that of p16 (r= -0.348, P= 0.044). In stepwise regression and the best subset regression, the expression of p16 (P=0.034) and phosphorylated Rb(Ser795) (P= 0.030) were the only determinants of the mitotic index. Conclusion The expression of phosphorylated Rb(Ser795) could be considered as a mark of the interaction between p16 and cyclin D1. The detection of phosphorylated Rb, p16 and cyclin D1 will be possibly helpful to the oncogenesis investigation on the esophageal carcinoma.

PAK4在细胞骨架和细胞周期调控中的作用研究进展

p21活化蛋白激酶(p21-activated protein kinase,PAK)是一类高度保守的丝氨酸/苏氨酸蛋白家族,是Rho家族小GTP酶的效应蛋白,参与多种信号通路传导。PAK4是PAK家族中最有代表性的成员,通过磷酸化下游底物,调控细胞骨架重组、细胞增殖和细胞周期进展等。PAK4过表达见于胰腺癌、胃癌和卵巢癌等多种肿瘤,参与肿瘤发生和肿瘤迁移等过程。研究PAK4的结构、作用机制对于揭示细胞周期调控和肿瘤生物学行为有重要价值。本文阐述和归纳了PAK4的结构、激活过程及其在细胞骨架、细胞周期进展中的生物学作用,并综述了PAK4调控妇科肿瘤发生发展以及PAK4抑制剂的最新研究进展。

ubr1基因缺失对粟酒裂殖酵母有丝分裂动力学的影响

为探究粟酒裂殖酵母(Schizosaccharomyces pombe)中ubr1基因缺失后有丝分裂动力学的变化情况,采用荧光蛋白标记和活细胞成像技术分析ubr1基因缺失型(ubr1Δ)菌株和野生型(WT)菌株细胞形态和有丝分裂期的细胞动力学差异。结果显示,ubr1基因缺失后细胞长宽比变小,微管束数目增多,细胞形态出现短棒状和梨形。有丝分裂过程中18.5%的ubr1Δ菌株会错误形成双动粒,且有丝分裂后期纺锤体伸长长度较野生型菌株增加(1.58±0.76)μm,有丝分裂后期持续时间较野生型菌株延长(3.00±1.36)min。有丝分裂末期纺锤体出现鱼钩型和S型断裂,且纺锤体断裂延迟。同时,ubr1Δ菌株与野生型菌株相比肌动蛋白环初始直径增加(1.12±0.19)μm,形成时间延长(3.60±2.85)min,收缩时间延长(4.90±0.21)min,但收缩速度并无明显差异。研究结果可为明确ubr1基因在有丝分裂中的功能及分子机制提供科学依据。

中间体残体与初级纤毛的关系及其在肿瘤生长中的作用

最新研究表明,初级纤毛细胞外发生途径与一种特殊细胞器—中间体残体相关。中间体残体是细胞有丝分裂末期中间体脱落形成的点状细胞器,中间体残体形成后在细胞表面移动,当与中心体在细胞表面靠近后,调节中心体在其母中心粒的顶端形成初级纤毛。初级纤毛可作为抑制肿瘤发生的细胞器,并在多种肿瘤中丢失,研究发现中间体残体在肿瘤细胞中聚集影响初级纤毛的发生。更重要的是,中间体残体和初级纤毛在肿瘤细胞中均通过自噬途径降解,中间体残体还可以将肿瘤信号通路因子传递给初级纤毛进而影响肿瘤细胞的发生发展。本文综述了中间体残体和初级纤毛的基本结构及形成过程,详细阐述了中间体残体调控初级纤毛发生的具体机制,探讨及展望了中间体残体调控初级纤毛形成在肿瘤发生发展和靶向治疗中的作用及研究趋势。

镉和氧化石墨烯对大蒜根尖细胞的影响

为了探究氧化石墨烯(GO)对大蒜根尖细胞抗镉(Cd)胁迫能力的影响,以大蒜为实验材料,研究了Cd和GO单一及共处理对大蒜生长、根尖细胞活性、根尖分生区细胞核仁形态、有丝分裂及Cd的亚细胞分布的影响.结果显示:①在本研究设置的浓度下Cd胁迫会显著抑制大蒜生长,GO会显著促进大蒜生长,GO和Cd共同处理则会在一定程度上缓解Cd对大蒜的生长抑制作用;②在单一Cd胁迫下,Cd对大蒜根尖细胞活性有毒害作用,且抑制作用随时间延长而加重,GO对Cd胁迫下大蒜根尖细胞活性下降有明显的缓解作用;③GO和Cd均会影响大蒜根尖细胞的有丝分裂并导致染色体畸变,二者共存时,GO会在一定程度上缓解Cd对大蒜有丝分裂的抑制作用;④Cd胁迫对大蒜根尖分生区细胞核仁形态有毒害作用,可以诱导核仁物质缓慢向外流出到核质甚至到细胞质基质,GO能有效缓解Cd胁迫对核仁结构的毒害作用,抑制了核仁物质向外流出;⑤GO可通过调节Cd的亚细胞分布在一定程度上缓解Cd对大蒜根部细胞的损伤.

细胞分裂周期蛋白73基因缺失抑制粟酒裂殖酵母细胞的有性生殖和有丝分裂

cdc73基因编码粟酒裂殖酵母(Schizosaccharomyces Pombe)RNA聚合酶Ⅱ辅助因子Cdc73,参与G_(2)期检查点激活并调控细胞周期。但cdc73基因缺失对细胞有丝分裂动力学的调控尚不清楚。本研究采用活细胞成像、荧光蛋白标记技术探究粟酒裂殖酵母cdc73基因缺失后对细胞有性生殖和细胞有丝分裂中微管、肌动蛋白、线粒体和组蛋白动力学的影响。结果表明:在有性生殖中,cdc73基因缺失会导致子囊孢子长度增加14.23%,产4个孢子数量的细胞减少64.08%;活细胞成像结果分析发现,在有丝分裂中,分裂后期微管伸长的长度缩短11.21%,伸长时间减少17.39%;肌动蛋白环形成和收缩速率分别降低33.33%和26.09%,形成和收缩时间分别延长58.00%和40.38%;同时,肌动蛋白环、线粒体和组蛋白表达量都增加。本研究揭示了cdc73基因缺失可抑制有丝分裂中纺锤体的伸长,延缓肌动蛋白环的形成与收缩,为进一步探寻Cdc73蛋白在细胞分裂中参与调控微管和肌动蛋白动力学功能提供了一定的科学依据。

人结直肠癌细胞周期各时相中双微体存在形式及复制分离时期的分析

目的探究双微体(Double minute chromosomes,DMs)在肿瘤细胞周期各时相内的主要存在形式,明确DMs的复制、分离时相。方法通过无血清饥饿法阻滞人结直肠癌细胞COLO 320DM周期进展后,取花萼海绵体诱癌素A(Calyculin-A)诱导间期细胞以染色质超前凝集制备核型样本。用秋水仙素处理COLO 320DM细胞以获取有丝分裂(M)期细胞进行核型分析。在普通光学显微镜下观察并拍摄DNA合成前期(G1期)、DNA合成后期(G2期)、M中期、M后期和M末期核型,并统计DMs数量。结果DMs在G1期、M后期和M末期时相细胞中主要以单体形式存在;在G2期和M中期时相细胞中主要以双体形式存在。结论单体型DMs在S期发生复制后由单体转变为双体,双体型DMs在M后期完成分离并由双体转变为单体。

SIRT2在小鼠1-细胞期受精卵中的表达和定位研究

目的:探讨沉默信息调节蛋白2(SIRT2)在小鼠1-细胞期受精卵的表达和定位。方法:利用qRT-PCR、Western blotting分别检测SIRT2在小鼠1-细胞期受精卵发育全过程中mRNA和蛋白表达谱;采用细胞免疫荧光法观察SIRT2和α-tubulin的共定位情况。结果:小鼠1-细胞期受精卵SIRT2 mRNA和蛋白表达变化趋势一致,均由G 1向G 2期过渡时表达水平升高;由G 2期向M期过渡时表达水平下降(P<0.05)。在G 2期向M期过渡过程中,SIRT2的定位由细胞质向细胞核转移,于M期中期进入细胞核并定位于纺锤体,在M期后期重新定位于细胞皮质。结论:SIRT2蛋白在小鼠1-细胞期受精卵中的表达呈细胞周期时相依赖性,在1-细胞期小鼠受精卵有丝分裂间期(G 1、S、G 2)中细胞质内表达增加,积累的SIRT2在某种条件下进入细胞核调节纺锤体微管动力学,在G 2/M过渡期发挥作用。

Research Progress on Targets and Selective Inhibitors of Polo-like Kinase-1(PLK-1)

In this paper,the biological function of PLK-1,the correlation between PLK-1 and tumors,and the latest research progress on PLK-1 inhibitors under study are reviewed,in order to provide references for the research and development of PLK-1 inhibitors.

Potential regulatory mechanism and clinical significance of synaptotagmin binding cytoplasmic RNA interacting protein in colorectal cancer

BACKGROUND Colorectal cancer(CRC)causes many deaths worldwide.Synaptotagmin binding cytoplasmic RNA interacting protein(SYNCRIP)is an RNA-binding protein that plays an important role in multiple cancers by epigenetically targeting some genes.Our study will examine the expression,potential effect,biological function and clinical value of SYNCRIP in CRC.AIM To examine the expression,potential effect,biological function and clinical value METHODS The expression of SYNCRIP was examined by immunohistochemistry arrays and high-throughput data.The effect of SYNCRIP gene in CRC cell growth was evaluated by CRISPR-Cas9 technology.The target genes of SYNCRIP were calculated using various algorithms,and the molecular mechanism of SYNCRIP in CRC was explored by mutation analysis and pathway analysis.The clinical value of SYNCRIP in prognosis and radiotherapy was revealed via evidence-based medicine methods.RESULTS The protein and mRNA levels of SYNCRIP were both highly expressed in CRC samples compared to nontumorous tissue based on 330 immunohistochemistry arrays and 3640 CRC samples.Cells grew more slowly in eleven CRC cell lines after knocking out the SYNCRIP gene.SYNCRIP could epigenetically target genes to promote the occurrence and development of CRC by boosting the cell cycle and affecting the tumor microenvironment.In addition,CRC patients with high SYNCRIP expression are more sensitive to radiotherapy.CONCLUSION SYNCRIP is upregulated in CRC,and highly expressed SYNCRIP can accelerate CRC cell division by exerting its epigenetic regulatory effects.In addition,SYNCRIP is expected to become a potential biomarker to predict the effect of radiotherapy.

BmN细胞有丝分裂及其染色体的研究

BmN细胞是家蚕分子遗传学、细胞工程、基因工程和杆状病毒表达系统中广泛应用的家蚕细胞,但其遗传学和细胞生物学背景知之甚少。用空气干燥法和培养细胞贴片法对BmN细胞的染色体和有丝分裂进行研究发现:BmN细胞是高度异倍化、高度多倍化的细胞系,其典型二倍体极少而高度多倍化群体达90％,最多的一群为533条染色体;BmN细胞有丝分裂以正常有丝分裂为主,但同时存在多倍核有丝分裂、核内有丝分裂及类无丝分裂等现象。

盐碱胁迫对油菜种子萌发和根尖细胞有丝分裂的影响

以冬油菜品种"陇油6号"为材料,通过中性盐NaCl∶Na2SO4(9 mol∶1 mol)混合胁迫(浓度为40、80、120、160、200 mmol·L-1和240 mmol·L-1,Na+浓度依次为44、88、132、176、220 mmol·L-1和264 mmol·L-1)和碱性盐NaHCO3∶Na2CO3(9 mol∶1 mol)混合胁迫(浓度为10、20、30、40、50 mmol·L-1和60 mmol·L-1,Na+浓度依次为11、22、33、44、55 mmol·L-1和66 mmol·L-1)处理油菜种子和幼苗,研究不同盐、碱胁迫下油菜种子萌发特性并观察根尖细胞有丝分裂的形态变化。结果表明:对"陇油6号"种子萌发和幼苗生长无害的最大混合盐浓度为160 mmol·L-1(pH=6.67,Na+=176 mmol·L-1),最大混合碱浓度为50 mmol·L-1(pH=9.29,Na+=55 mmol·L-1);随着盐、碱胁迫浓度增大,"陇油6号"幼苗根尖细胞有丝分裂指数逐渐降低,细胞微核率和染色体畸变率先上升后下降,碱性盐溶液胁迫下降幅度明显大于中性盐溶液胁迫;在同一盐、碱胁迫浓度时,处理时间越长,根尖细胞有丝分裂指数越低,染色体畸变率和微核率越高。这表明盐、碱胁迫对"陇油6号"种子萌发和幼苗生长是两种不同的胁迫,且碱性盐胁迫强于中性盐。

预处理对辣椒根尖细胞染色体制片的影响

该实验以中椒四号辣椒为实验材料,用常规压片法制备辣椒根尖细胞染色体标本.研究了短时间温度预处理对辣椒根尖细胞有丝分裂活性的影响,并比较了不同的预处理剂和冰水预处理时间对中期分裂相的影响.结果表明,经28℃非离体升温处理3h,冰水预处理24h、辣椒根尖可以获得数目较多、分散均匀的辣椒根尖细胞染色体早中期分裂相.

神经干细胞有丝分裂过程中nestin表达变化

在哺乳动物胚胎早期发育过程中 ,中枢神经系统的神经干细胞 ( NSCs)首先通过对称分裂进行自身复制 ,然后通过不对称分裂产生神经元和胶质细胞的前体细胞 ,这些细胞都表达一种中间纤维蛋白 ( IF) -神经巢蛋白 ( nestin)。nestin的表达是暂时的 ,一旦前体细胞分化成终末分化细胞如神经元或胶质细胞 ,nestin则被其他中间纤维蛋白所取代。不同中间纤维在体内不同时期表达的原因和执行的功能还不清楚 ,但可以肯定的是 nestin与 NSCs的多潜能性有关。本实验通过缩时显微成像和免疫组织化学技术观察了 NSCs在对称分裂过程中 nestin的表达变化 ,发现 nestin在前、中、后、末四期的荧光强度表达有不同程度的变化 ,分裂前期与分裂间期相同 ,前中期荧光强度有明显增强 ,到中期荧光强度反而有些下降 ,到末期荧光强度达到最大值 ,胞质分裂后荧光强度逐渐下降。这种波动表达现象将有助于揭示

小鼠单纯肝切除后再生肝组织内大小核分裂相和卵圆细胞的观察

目的:探究C57小鼠单纯2/3肝切除后肝组织内与再生和肝干细胞有关的病理形态学变化及其意义。方法:在4个对照组(正常组、假手术组、倒千里光碱组和倒千里光碱2/3肝切除组)的参照下,观察实验组C57小鼠单纯2/3肝切除后不同时间肝脏病理形态变化(核分裂相和卵圆细胞等)和CK19、AFP免疫组化检测情况。结果:单纯2/3肝切除后小鼠均出现不同程度的成熟肝细胞通过大核分裂相分裂增生、轻重不等的小胆管/终末胆管增生反应和个别例中有类似于倒千里光碱肝切除模型组的卵圆细胞增生;尤其还有大小核分裂相分布等一些文献未提及的形态学改变。结论:本研究提出肝流域假说的初步设想:肝内成体干细胞存在于小胆管/终末胆管附近,是肝主质细胞的重要源头;在其向中央静脉方向的流向上产生各级分化程度不同的子代(自卵圆细胞到小肝细胞、成熟肝细胞),沿肝板形成该流域干流;而经由血流到达肝脏的过客性干细胞可不同程度地在不同区段作为该流域的支流汇入干流并转分化为肝系细胞。