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Decreased Pollen Viability and Thicken Pollen Intine in Antisense Silenced Brassica campestris Mutant of BcMF19

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Journal of Integrative Agriculture 2014年第5期13卷 954-962页

作者： LIU Jin-long GAO Ming-hui LIU Ying CAO Jia-shu laboratory of cell and Molecular biology Institute of Vegetable Science Zhejiang University

Brassica campestris male fertility 19 （BcMF19; GenBank accession number GQ902048.1）, a gene that is specially expressed in tapetum cells and microspores during anther development in B. campestris ssp. chinensis, which is learned from the previous in situ hybridization study. In the present study, we constructed antisense-silenced plants of BcMF19 using B. campestris ssp. chinensis to validate this prediction. The morphology of the pistils, long anthers, and short anthers was significantly affected in 35sbcmf19 compared with the control samples. 4＇-6-Diamidino-2-phenylindole staining revealed that two generative nuclei and one large vegetative nucleus were not affected in the mutant compared with control. Statistical analysis of Alexander＇s staining results showed that 96% of the control pollen grains had vitality, whereas only 86% of the mutant pollen grains did. Under scanning electron microscopy, the mutant demonstrated numerous abnormal pollen grains and resembled dried persimmon. The frequency of normal pollen grains was approximately 18%. Under transmission electron microscopy, the pollen intine during the binucleate and mature pollen stages in 35sbcmf19 exhibited abnormal thickening, especially at the germinal furrows, compared with control. In vitro pollen germination test showed that the tips of the mutant pollen tubes transformed into globular alveoli and stopped growing compared with control. On the other hand, in vivo pollen germination test suggested that BcMF19 affected the pollen tube extension in the pistil. These findings indicate that BcMF19 is essential to the pollen development and pollen tube extension orB. campestris ssp. chinensis.

关键词： Brassica rapa ssp. chinensis PGIP pollen polygalacturonase-inhibiting protein tapetum pollen germination

Effects of Huangqi (Hex) on Inducing cell Differentiation and cell Death in K562 and HEL cells

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Acta Biochimica et Biophysica Sinica 2004年第3期36卷 211-217页

作者： Xiao-Dong CHENG1, Chun-Hui HOU1, Xue-Jun ZHANG2, Heng-Yue XIE1, Wei-Ying ZHOU1, Lei YANG2, Shu-Bing ZHANG1, and Ruo-Lan QIAN1 State Key laboratory of Molecular biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China laboratory of Molecular cell biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences,320 Yueyang Road Shanghai 200031,China the Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences the Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China

Huangqi (Astragalus membranaceus), a traditional Chinese medicine, has been used to ameliorate side effects of cancer chemotherapy in China. However, little is known about its molecular mechanisms. Here we show that induction of K562 or HEL cells with 1.5 mg/ml of Huangqi (Hex) (Components extracted from Huangqi) for 3–5 d results in the expression of β-globin gene in both cell lines and leads to terminal differentiation. Moreover, the apoptosis in HEL cells can be induced by increasing concentration of Huangqi (Hex) to 4.5 mg/ml for 3–5 d. Upregulation of Apaf-1, caspase-3 and acetylcholinesterase (AChE) in HEL cells may play a crucial role in the process of apoptosis. The prospect of inducing expression of adult (β) globin gene and apoptosis selectively in cancer cells is obviously attractive from a therapeutic point of view.

关键词： cell death and differentiation Huangqi (Hex) human erythroleukemia cell lines

LKB1 in lung cancerigenesis:a serine/threonine kinase as tumor suppressor

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Protein & cell 2011年第2期2卷 99-107页

作者： Yijun Gao Gaoxiang Ge Hongbin Ji laboratory of Molecular cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China State Key laboratory of Molecular biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

Lung cancer is featured with high mortality,with a 15%five-year survival rate *** alterations,such as loss of function of tumor suppressor genes,frequently contribute to lung cancer initiation,progression and *** kinase B1(LKB1),as a serine/threonine kinase and tumor suppressor,is frequently mutated and inactivated in non-small cell lung cancer(NSCLC).Recent studies have provided strong evidences that LKB1 loss promotes lung cancerigenesis process,especially lung cancer progression and *** review will summarize recent progress on how LKB1 modulates the process of lung cancerigenesis,emphasizing on LKB1 downstream signaling pathways and biological *** will further discuss the potential development of prognostic biomarkers or therapeutic targets in lung cancer clinic based on the molecular alteration associated with deregulated LKB1 signaling.

关键词： liver kinase B1 lung cancer tumor suppressor

Cleavage of Supercoiled Circular Double-stranded DNA Induced by a Eukaryotic Cambialistic Superoxide Dismutase from Cinnamomum camphora

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Acta Biochimica et Biophysica Sinica 2004年第9期36卷 609-617页

作者： Bao-Zhong WANG, Xu-Bin WEI&, and Wang-Yi LIU* State Key laboratory of Molecular biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences, Shanghai 200031, China State Key laboratory of Molecular biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences

A eukaryotic cambialistic superoxide dismutase (SOD) has been purified to homogeneity from mature seeds of the disease- and insect-resistant camphor tree (Cinnamomum camphora). Besides the known role of this SOD in protecting cells against oxidative stress, it can induce the cleavage of supercoiled double-stranded DNA into nicked and linear DNA. It can not cleave linear DNA or RNA, demonstrating there is no DNase or RNase in the purified cambialistic SOD. Furthermore, the SOD can linearize circular pGEM-4Z DNA that is relaxed by topoisomerase I. This result indicates that the DNA-cleaving activity re- quires substrates being topologically constrained. The supercoiled DNA-cleaving activity of the cambialistic SOD can be inhibited by either SOD inhibitor (azide) or catalase and hydroxyl radical scavengers (ethanol and mannitol). The chelator of iron, diethylenetriaminepentaacetic acid (DTPA), also inhibits the supercoiled DNA-cleaving activity. These results show that the dismutation activity is crucial for the supercoiled DNA cleavage. The modification of tryptophan residue of the cambialistic SOD with N-bromosuccinimide (NBS) shows that these two activities are structurally correlative. The reaction mechanism is proposed that the hydroxyl radical formed in a transition-metal-catalyzing Fenton-type reaction contributes to the DNA- cleaving activity. In addition, the cleavage sites in supercoiled pGEM-4Z DNA are random.

关键词： cambialistic SOD Fenton-type reaction hydroxyl radical relaxed circular duplex DNA supercoiled DNA cleavage

NUCLEAR PORE COMPLEX IN SPERMATOGENESIS OF BEIJING DOMESTIC DUCK——ELECTRON MICROSCOPE OBSERVATION OF FREEZE-ETCHING

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Science China Chemistry 1984年第3期27卷 265-272页

作者：黄宗平汪德耀李文镇李向印 Hebei Medical College Department of biology Xiamen University laboratory of Basic Medival Science laboratory of cell biology

The nuclear pore complex in spermatogenesis of Beijing domestic duck has been studiedby means of electron microscopic observation of freeze-etching. The structure of the poreobserved proves to be in accord with the model of the nuclear pore complex proposed respec-tively by Franke (1970) and Roberts et al. (1970). But its central granule may be present(1--3 granules) or absent. The granules are considered to be material in transit. The sizeand frequency of the pore vary with different types of reproductive cells, which is believedto be related to the cells’ function. The pores concentrate on the side of nucleus near Golgizone in primary spermatocyte and near the proacrosome of spermatid, and this shows thefunctional variety of parts in the nucleus.

关键词： ELECTRON MICROSCOPE OBSERVATION OF FREEZE-ETCHING NUCLEAR PORE COMPLEX IN SPERMATOGENESIS OF BEIJING DOMESTIC DUCK

Hedgehog in the Drosophila testis niche:what does it do there?

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Protein & cell 2013年第9期4卷 650-655页

作者： Zhao Zhan Chenyu Pan Yun Zhao State Key laboratory of cell biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China

Stem cell niche is a specialized microenvironment crucial to *** testis in Drosophila contains two dif-ferent types of stem cells,the germline stem cells and the somatic cyst stem cells that are sustained by their respec-tive niche signals,thus is a good system for studying the interaction between the stem cells and their hosting *** JAK-STAT and BMP pathways are known to play critical roles in the self-renewal of different kinds of stem cells,but the roles of several other pathways have emerged recently in a complex signaling network in the testis *** of independent observations from three research groups have uncovered an important role of Hedgehog(Hh)in the Drosophila testis *** this review,we summarize these recent fi ndings and discuss the interplay between the Hh signaling mechanisms and those of the JAK-STAT and BMP *** also dis-cuss directions for further investigation.

关键词： Drosophila testis stem cell Hedgehog JAK-STAT

Impaired reproduction in transgenic mice overexpressing γ-aminobutyric acid transporter Ⅰ (GAT1)

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cell Research 2004年第1期14卷 54-59页

作者： Jia Hua HU~(1,*), Jin Fu ZHANG~(2,*), Ying Hua MA~1, Jie JIANG~1, Na YANG~1, Xin Bo LI~3, Zhi Guang YU CHI~4, Jian FEI~(1,5,**), Li He GUO~(1,**) ~1laboratory of Molecular cell biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China. ~2Department of Urology, Tongji Hospital, Tongji University, 389 Xincun Road, Shanghai 200065, China. ~3New Drug R & D Center, North China Pharmaceutical Group Corporation, 388 Heping East Road, Shijia Zhuang 050015, Hebei, China. ~4School of life sciences, Fudan University, 220 Handan Road, Shanghai 200433, China. ~5Shanghai Research Center for Biomodel Organism, Shanghai 201203, China. laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China.

It is well documented that γ-aminobutyric acid (GABA) system existed in reproductive organs. Recent researches showed that GABA_A and GABA_B receptors were present in testis and sperm, and might mediate the acrosome reaction induced by GABA and progesterone. GABA transporter Ⅰ (GAT1) also existed in testis and sperm, but its physiological function was unknown. In the present study, we used GAT1 overexpressing mice to explore GAT1 function in male reproductive system. We found that the expression level of GAT1 continuously increased in wild-type mouse testis from 1 month to 2 months after birth. GAT1 overexpression in mouse affected testis development, which embodied reduced testis mass and slowed spermatogenesis in transgenic mice. Moreover, transgenic mice showed increase of the percentage of broken sperm. The further study revealed that the reproductive capacity was impaired in GAT1 overexpressing mice. In addition, testosterone level was significantly low in transgenic mice compared with that in wi

关键词： GABA GAT1 testes sperm reproduction transgenic.

Mitosin/CENP-F is a conserved kinetochore protein subjected to cyto plasmic dynein-mediated poleward transport

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cell Research 2003年第4期13卷 275-283页

作者： ZHEN YE YANG, JING GUO, NING LI, MIN QIAN, SHENG NIAN WANG, XUE LIANG ZHUlaboratory of Molecular cell biology, Institute of Biochemistry and cell biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China 1laboratory of Molecular cell biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences 320 Yueyang Road Shanghai 200031 China

Mitosin/CENP-F is a human nuclear protein transiently associated with the outer kinetochore plate in M phase and is involved in M phase progression. LEK1 and CMF1, which are its murine and chicken orthologs, however, are implicated in muscle differentiation and reportedly not distributed at kinetochores. We therefore conducted several assays to clarify this issue. The typical centromere staining patterns were observed in mitotic cells from both human primary culture and murine, canine, and mink cell lines. A C-terminal portion of LEK1 also conferred centromere localization. Our analysis further suggests conserved kinetochore localization of mammalian mitosin orthologs. Moreover, mitosin was associated preferentially with kinetochores of unaligned chromosomes. It was also constantly transported from kinetochores to spindle poles by cytoplasmic dynein. These properties resemble those of other kinetochore proteins important for the spindle checkpoint, thus implying a role of mitosin in this checkpoint. The

关键词： mitosin LEKl CENP-F kinetochore.

Effects of PKCa antisense RNA on human breast cancer cell proliferation and expression of cyclinD1 and CDK4

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Chinese Science Bulletin 1999年第12期44卷 1110-1114页

作者： SANG Jianli WANG Yongchaolaboratory of cell biology , Department of biology, Beijing Normal University, Beijing 100875 , China laboratory of cell biology Department of Biology Beijing Normal University Beijing China

The role of PKCα in human breast cancer cell proliferation and expression of cyclinD1 and CDK4 has been investigated using inhibition of PKCα expression by its antisense RNA. When PKCα expression was inhibited the rate of cell proliferation decreased apparently and the levels of cyclinD1 and CDK4 mRNA were lower than the control. The results showed that PKCα, a key member of signal transduction system, played an important role in human breast cancer cell proliferation and had a close relationship with expression of cyclinD1 and CDK4 which control start of cell cycle.

关键词： breast cancer PKCα cell proliferation cyclinD1 CDK4.