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Suppression of Type 1 Insulin-like Growth Factor Receptor Expression by Small Interfering RNA Inhibits A549 Human Lung Cancer Cell Invasion in vitro and Metastasis in Xenograft Nude Mice

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Acta Biochimica et Biophysica Sinica 2007年第2期39卷 137-147页

作者： Jianfang QIAN~1 Aiqiang DONG~1 Minjian KONG~1 Zhiyuan MA~2 Junqiang FAN~1 and Guanyu JIANG~3 ~1department of Cardiothoracic Surgery,second affiliated hospital,school of medicine,zhejiang university,hangzhou 310009,china ~2department of Cardiovascular Surgery,Shanghai Jiaotong university affiliated First People's hospital,Shanghai 200080,china ~3department of emergency,second affiliated hospital school of medicine,zhejiang university,hangzhou 310009,china department of Cardiothoracic Surgery Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China department of Cardiovascular Surgery Shanghai Jiaotong University Affiliated First People's HospitalShanghai 200080 China department of emergency Second Affiliated Hospital School of Medicine Zhejiang UniversityHangzhou 310009 China

Cancer invasion and metastasis, involving a variety of pathological processes andcytophysiological changes,contribute to the high mortality of lung *** type 1 insulin-like growthfactor receptor (IGF-1R),associated with cancer progression and invasion,is a potential anti-invasion andanti-metastasis target in lung *** inhibit the invasive properties of lung cancer cells,we successfullydown-regulated IGF-1R gene expression in A549 human lung cancer cells by small interfering RNA (siRNA)technology,and evaluated its effects on invasion-related gene expression,tumor cell in vitro invasion,andmetastasis in xenograft nude mice.A549 cells transfected with a plasmid expressing hairpin siRNA forIGF-1R showed a significantly decreased IGF-1R expression at the mRNA level as well as the *** biological assays,transfected A549 cells showed a significant reduction of cell-matrix adhesion,migration and *** with these results,we found that down-regulation of IGR-1Rconcomitantly accompanied by a large reduction in invasion-related gene expressions,including MMP-2,MMP-9,u-PA,and IGF-1R specific downstream *** tail vein injections of plasmid expressinghairpin siRNA for IGF- 1R significantly inhibited the formation of lung metastases in nude *** resultsshowed the therapeutic potential of siRNA as a method for gene therapy in inhibiting lung cancer invasionand metastasis.

关键词： lung cancer invasion metastasis type 1 insulin-like growth factor receptor small interfering RNA

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Shp2,a novel oncogenic tyrosine phosphatase and potential therapeutic target for human leukemia

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Cell Research 2007年第4期17卷 295-297页

作者： Rongzhen Xu~1 ~1department of Hematology,second affiliated hospital,school of medicine,zhejiang university,hangzhou 310009,china department of Hematology Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Shp2,encoded by the PTPN11 gene in human,is aubiquitously expressed protein tyrosine phosphatase thatcontains two N-terminal Src homology 2(SH2)domains(N-SH2,C-SH2,respectively),a catalytic protein-tyrosinephosphatase(PTP)domain,and a C-terminal tail withtyrosyl phosphorylation sites and a prolyl-rich motif[1].The progress of our understanding of biological functionsof Shp2 has clearly shown that Shp2 plays an importantrole not only in biology of normal hematopoietic cells andother mammalian cells,but also in the development ofleukemia and other *** recently,PTPNll genehas been firmly established as the first proto-oncogene thatencodes a protein tyrosine phosphatase[1-3].In the hema-topoietic system,most if not all function of Shp2 is to actas a positive component that is essential for proliferationand/or survival of hematopoietic cells through regulationof signaling pathways involving Erk,Akt and STAT5[1-4].Over the past few years,a number of disease-associatedShp2 mutants have been identified in human leukemia andother malignancies[1,3,4].Recently,studies from ourlaboratories and others strongly suggest that dysregula-tion of wild-type Shp2 enzyme may be involved in the

关键词： SHP Shp2,a novel oncogenic tyrosine phosphatase and potential therapeutic target for human leukemia CML

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Distribution of cysteinyl leukotriene receptor 2 in human traumatic brain injury and brain tumors

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Acta Pharmacologica Sinica 2005年第6期26卷 685-690页

作者： Hua HU~(2,3), Gao CHEN~2, Jian-min ZHANG~2, Wei-ping ZHANG~3, Lei ZHANG~3, Qiu-fu GE~3, Hong-tian YAO~4, WeiDING~4, Zhong CHEN~3, Er-qing WEI~(3,5) ~2 department of Neurosurgery, the second affiliated hospital, school of medicine, zhejiang university, hangzhou 310009 ~3 department of Pharmacology school of medicine, zhejiang university, hangzhou 310031 ~4 department of Pathology, the First affiliated hospital, school of medicine, zhejiang university, hangzhou 310003, china department of Neurosurgery the Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China department of Pharmacology School of Medicine Zhejiang University Hangzhou China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim: To determine the distribution of cysteinyl leukotriene receptor 2 （CysLT2）, one of the cysteinyl leukotriene receptors, in human brains with traumatic injury and tumors. Methods: Brain specimens were obtained from patients who underwent brain surgery. CysLT2in brain tissues was examined using immunohis-tochemical analysis. Results: CysLT2was expressed in the smooth muscle cells （not in the endothelial cells） of arteries and veins. CysLT2was also expressed in the granulocytes in both vessels and in the brain parenchyma. In addition, CysLT2was detected in neuron- and glial-appearing cells in either the late stages of traumatic injury or in the area surrounding the tumors. Microvessels regenerated 8 d after trauma and CysLT2expression was recorded in their endothelial cells. Conclusion: CysLT2is distributed in vascular smooth muscle cells and granulocytes, and brain trauma and tumor can induce its expression in vascular endothelial cells and in a number of other cells.

关键词： cysteinyl leukotriene receptor 2 brain injuries brain neoplasms vascular smooth muscle cell

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Effects of CpG oligodeoxynucleotide on transcription factors GATA-3 and T-bet mRNA expression in asthmatic mice

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Acta Pharmacologica Sinica 2005年第9期26卷 1117-1122页

作者： Zu-qun WU~(1,2) Yi-ping XU~3 Hua XIANG~4 Hua-hao SHEN~11 department of Respiratory medicine,the second affiliated hospital,4 department of Pathology,the First affiliated hospital,school of medicine,zhejiang university,hangzhou 310009,china 3 department of Respiratory medicine,Renji hospital,Shanghai second Medical university Shanghai 200001,china department of Respiratory medicine the Second Affiliated Hospital Hangzhou China department of Respiratory medicine Renji Hospital Shanghai Second Medical University Shanghai China department of Pathology the First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

Aim:To investigate effects ofCpG oligod'eoxynucleotide(CpG ODN)on the mRNAexpression of transcription factors GATA binding protein 3(GATA-3)and T-boxexpressed in T cells(T-bet)in asthmatic ***:An asthmatic mousemodel was established and treated with CpG *** inflammatory cells andeosinophils in bronchoalveolar lavage fluid(BALF)were counted and inflamma-tory cell infiltration in lung tissue was ***-y and interleukin-4concentrations in BALF and splenocyte culture supernatants were detected us-ing an enzyme-linked immunosorbent *** factor GATA-3 and T-bet mRNA expression in splenocytes and lung tissue were detected by reversetranscription-polymerase chain ***:Total inflammatory cells andeosinophils in BALF were reduced in the CpG ODN-treated group compared withthe asthma group,and inflammatory cell infiltration in lung tissue was also signifi-cantly *** ODN treatment increased the interferon-γ concentrationbut decreased the interleukin-4 concentration in both BALF and splenocyte cul-ture ***-3 mRNA expression was reduced in both lung tissueand splenocytes in the CpG ODN-treated group,while the mRNA ratio ofT-bet toGATA-3 in splenocytes was ***:CpG ODN treatment inhibitsairway inflammatory cell infiltration and regulates interferon-y/interleukin-4 syn-thesis in asthmatic mice,possibly through a mechanism of downregulation ofGATA-3 mRNA expression in both lung tissue and splenocytes.

关键词： asthma inflammation CpG oligodeoxy-nucleotide GATA-3 T-bet mice

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Parthenolide protects human lens epithelial cells from oxidative stress-induced apoptosis via inhibition of activation of caspase-3 and caspase-9

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Cell Research 2007年第6期17卷 565-571页

作者： Hangping YaoXiajing TangXueting ShaoLei FengNanping WuKe YaoNational Key Laboratory of Infectious Diseases, Institute of Infectious Diseases, First affiliated hospital, school of medicine, zhejiang university, hangzhou, chinaEye Center, second affiliated hospital school of medicine, zhejiang university, 88 Jiefang Road, hangzhou 310009, china National Key Laboratory of Infectious Diseases Institute of Infectious Diseases First Affiliated Hospital School of Medicine Zhejiang University Hangzhou China Eye Center Second Affiliated Hospital School of Medicine Zhejiang University Hangzhou China

The apoptosis of lens epithelial cells has been proposed as the common basis of cataract formation, with oxidative stressas the major cause. This study was performed to investigate the protective effect of the herbal constituent parthenolideagainst oxidative stress-induced apoptosis of human lens epithelial （HLE） cells and the possible molecular mechanismsinvolved. HLE cells （SRA01-04） were incubated with 50μM H2O2 in the absence or presence of different doses ofparthenolide （10, 20 and 50μM）. To study apoptosis, the cells were assessed by morphologic examination and AnnexinV-propidium iodide double staining flow cytometry; to investigate the underlying molecular mechanisms, the expressionof caspase-3 and caspase-9 were assayed by Western blot and quantitative RT-PCR, and the activities of caspase-3 andcaspase-9 were measured by a Chemicon caspase colorimetric activity assay kit. Stimulated with H2O2 for 18 h, a highfraction of HLE cells underwent apoptosis, while in the presence of parthenolide of different concentrations, dose-depen-dent blocking of HLE cell apoptosis was observed. The expression of caspase-3 and caspase-9 induced by H2O2 in HLEcells was significantly reduced by parthenolide both at the protein and mRNA levels, and the activation of caspase-3 andcaspase-9 was also suppressed by parthenolide in a dose-dependent manner. In conclusion, parthenolide prevents HLEcells from oxidative stress-induced apoptosis through inhibition of the activation of caspase-3 and caspase-9, suggestinga potential protective effect against cataract formation.

关键词： parthenolide human lens epithelial cells apoptosis caspase-3 and caspase-9

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Inhibitory effect of acetamide-45 on airway inflammation and phosphodiesterase 4 in allergic rats

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Acta Pharmacologica Sinica 2005年第12期26卷 1492-1496页

作者： Kai WANG~2 Hua-hao SHEN~(2,4) Jun-chun CHEN~2 Zhong CHEN~3 2 Respiratory department,the second affiliated hospital,Medical school of zhejiang university,hangzhou 310009 3 department of Pharmacology,Medical school of zhejiang university,hangzhou 310031,china Respiratory department the Second Affiliated Hospital Medical School of Zhejiang University Hangzhou China department of Pharmacology Medical School of Zhejiang University Hangzhou China

Aim:To determine the effects of acetamide-45 on respiratory function,airwayinflammation,and the activity of phosphodiesterase 4（PDE4）in allergic ***:Rats were sensitized by a single intramuscular injection with ovalbumin（OVA）and were challenged with ovalbumin applied by using an aerosol repeat-edly for 7 d after 2 ***-45 at concentrations of 5,10,or 30 mg/kg wasthen administered by intraperitoneal *** in dynamic lung compli-ance and lung resistance,the accumulation of inflammatory cells in bronchoalveolarlavage,PDE4 activity,and the concentration of interleukin-4 in rat lung tissuewere ***:Seven days of treatment with acetamide-45 preventedeosinophil accumulation in allergic *** doses of 5,10,and 30 mg/kg,aceta-mide-45 decreased lung resistance to 0.20±0.04,0.25±0.07,and 0.22±0.05cmH2O·s-1·mL-1,respectively（P39±0.06,and 0.42±0.09 mL/cmH20（P3±57,and 238±36 nmol·g-1·min-1（P3,5.95±1.20,and 5.68±2.20 μtg/g protein（P<0.05 vs OVA）.Conclusions:Acetamide-45 wasfound to improve respiratory function and inhibit airway inflammation in thisanimal model,and the PDE4 activity of lung tissue was obviously ***-45 was an effective anti-inflammatory agent in respiratory inflammation,and the mechanism of its action might depend on inhibition of PDE4.

关键词： acetamide-45 3’,5’-cyclic nucleotide phosphodiesterase respiratory function tests eosinophils asthma

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Effect of Porphyromonas gingivalis PrtC on cytokine expression in ECV304 endothelial cells and its level in subgingival plaques from patients with chronic periodontitis

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Acta Pharmacologica Sinica 2007年第7期28卷 1015-1023页

作者： Yan-min WU~2,Li-li CHEN~(2,6),Jie YAN~(3,6),Chun-yan ZHUANG~4,Zhi-yuan GU~5~2 department of Stomatology,the second affiliated hospital,College of medicine,zhejiang university,hangzhou 310009,china ~3 Depart-ment of Medical Microbiology and Parasitology,College of medicine,zhejiang university,hangzhou 310006,china ~4 department ofStomatology,the affiliated hospital of the school of medicine,Ningbo university,Ningbo 315020,china ~5 department of Oral andMaxillofacial Surgery,the affiliated hospital of Stomatology,College of medicine,zhejiang university,hangzhou 310006,china department of Stomatology the Second Affiliated Hospital College of Medicine Zhejiang University Hangzhou China department of Medical Microbiology and Parasitology College of Medicine Zhejiang University Hangzhou China department of Stomatology the Affiliated Hospital of the School of Medicine Ningbo University Ningbo China department of Oral and Maxillofacial Surgery the Affiliated Hospital of Stomatology College of Medicine Zhejiang University Hangzi China

Aim:To investigate the effect of the collagenase gene (prtC) product ofPorphyromonas gingivalis on inducing host cells to secrete inflammatorycytokines,and to discuss the correlation between the PrtC level in subgingivalplaque samples and clinical ***:A prokaryotic expression sys-tem pET32a-prtC-Escheria coli BL21 DE3 was *** and im-munoreactivity of the recombinant PrtC protein (rPrtC) was identified by *** was applied to detect interleukin (IL)-1α,IL-8,and TNF-α levelsin supernatants from rPrtC-induced human umbilical vein endothelial cells (HUVEC)originated ECV304 *** parameters recorded at baseline and after treat-ment included bleeding on probing (BOP),probing depth (PD),and attachmentloss (AL).ELISA was established to measure the PrtC level in 196 subgingivalplaque samples from 49 patients with chronic ***:After co-incubation with 1 μg/mL rPrtC for 24 h and with 5 or 10 μg/mL rPrtC for 12 h,thelevels of IL-1α,IL-8,and TNF-α secreted by the ECV304 cells increased signifi-candy (P6 mm PDsites was higher than that in the BOP-negative or the ≤2 mm AL or ≤6 mm PD sites(P6 mm PD sites,the PrtC levels changed insignifi-cantly (P>0.05).Conclusion:rPrtC is able to directly induce host cells to synthe-size and secrete IL-1α,IL-8,and TNF-α.The PrtC level in subgingival samples iscorrelated with BOP,AL,and PD.

关键词： Porphyromonas gingivalis collagenase cloning cytokine ELISA

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Human C-C chemokine receptor 3 monoclonal antibody inhibits pulmonary inflammation in allergic mice

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Acta Pharmacologica Sinica 2007年第11期28卷 1791-1796页

作者： Kai WANG~2,Hua-hao SHEN~2,Wen LI,Hua-qiong HUANGdepartment of Respiratory medicine,the second affiliated hospital,Medical school of zhejiang university,hangzhou 310009,china department of Respiratory medicine the Second Affiliated Hospital Medical School of Zhejiang University Hangzhou China

Aim:To evaluate the effect of C-C chemokine receptor 3 (CCR3) blockade onpulmonary inflammation and mucus production in allergic ***:Weused the synthetic peptide of the CCR3 NH2-terminal as the immunizing antigenand generated murine monoclonal antibody against the human *** addition,the generated antibody was administered to mice sensitized and challenged *** inflammatory cells in bronchoalveolar lavage,cytokine levels,pulmonary histopathology,and mucus secretion were ***:TheWestern blotting analysis indicated that the generated antibody bound to *** allergic mice treated with the antihuman CCR3 antibody exhib-ited a significant reduction of pulmonary inflammation accompanied with the al-teration of ***:The antibody we generated was specific *** inhibition of airway inflammation and mucus overproduction by theantibody suggested that the blockade of CCR3 is an appealing therapeutical tar-get for *** present research may provide an experimental basis for thefurther study of this agent.

关键词： asthma C-C chemokine interleukin-4 lungs inflammation

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Changes of phospholipase D activity of rat peritoneal mast cells in degranulation

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Acta Pharmacologica Sinica 2004年第1期 106-111页

作者： Yun- bi LU, Ming WU, Han-liang ZHOUdepartment of Pharmacology, school of medicine, zhejiang university, hangzhou 310031 department of Cardiovascular Thoracic Surgery, second affiliated hospital, school of medicine, zhejiang university, hangzhou 310009, china

IM: To study the changes of phospholipase D (PLD) activity of actively sensitized rat peritoneal mast cells (RPMC) in degranulation. METHODS: Degranulation of RPMC was determined by measurement of β-hexosaminidase release. PLD activity assay was carried out by measurement of PLD product, choline, with chemiluminescent oxidation of luminol. RESULTS: Actively sensitized RPMC challenged with ovalbumin (0.5-8 mg/L for 120 s, 4 mg/L for 15-120 s) resulted in significant activation of PLD accompanied with the increment of P-hexosaminidase release. PLD activity of sensitized RPMC was increased by more than 2-fold compared with that of unsensitized RPMC which contained low levels of PLD activity [(35±13) pmol choline/min in 1×106cells], but β-hexosaminidase releases of the sensitized cells were as low as spontaneous releases.

关键词： ovalbumin phospholipase D immunization mast cells

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What we know about ST13, a co-factor of heat shock protein, or a tumor suppressor?

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Journal of zhejiang university-Science B(Biomedicine & Biotechnology) 2007年第3期8卷 170-176页

作者： SHI Zheng-zheng, ZHANG Jia-wei, ZHENG Shu (The second affiliated hospital, Cancer Institute, school of medicine, zhejiang university, hangzhou 310009, china) The second affiliated hospital Cancer Institute School of Medicine Zhejiang University Hangzhou 310009 China.

This article is to summarize the molecular and functional analysis of the gene “suppression of tumorigenicity 13” (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone) of the 70-kDa heat shock proteins (Hsc/Hsp70). By collaborating with other positive co-factors such as Hsp40 and the Hsp70-Hsp90 organizing protein (Hop), or competing with negative co-factors such as Bcl2-associated athanogen 1 (Bag1), Hip facilitates may facilitate the chaperone function of Hsc/Hsp70 in protein folding and repair, and in controlling the activity of regulatory proteins such as steroid receptors and regulators of proliferation or apoptosis. Although the nomenclature of ST13 implies a role in the suppression of tumorigenicity (ST), to date available experimental data are not sufficient to support its role in cancer development, except for the possible down-regulation of ST13 in gastric and colorectal cancers. Further investigation of this gene at the physiological level would benefit our understanding of diseases such as endocrinological disorders, cancer, and neurodegeneration commonly associated with protein misfolding.

关键词： ST13 Hip p48 Hsc/Hsp70 Heat shock protein Protein folding Steroid receptor Tumorigenicity

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