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Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line

Acta Pharmacologica Sinica 2007年第11期28卷 1819-1826页

作者： Hong-zhen HU~2,Yue-bo YANG~3,Xiang-dong XU~4,Hong-wei SHEN~5,Yi-min SHU~6,Zi REN~7,Xiao-mao LI~3,Hui-ming SHEN~3,Hai-tao ZENG~(3,8)~2 department of Gynecology and Obstetrics,Yijishan Hospital,Wannan Medical College,Wuhu 241001,china ~3 department of Gynecologyand Obstetrics,the Third Affiliated Hospital,sun yat-sen university,guangzhou 510080,china ~4 department of surgery,the first affiliatedhospital,sun yat-sen university,guangzhou 510080,china ~2 department of Gynecology and Obstetrics,the first Affiliated Hospital,sunyat-sen university,guangzhou 510080.china ~6 department of Obstetrics and Gynecology,IVF Program,Stanford university MedicalCenter,Palo Alto,CA 94304,USA ~7 department of Gynecology and Obstetrics,Guangdong Provincial People's Hospital,guangzhou510080.china department of Gynecology and Obstetrics Yijishan Hospital Wannan Medical College Wuhu China department of Gynecology and Obstetrics the Third Affiliated Hospital Sun Yat-sen University Guangzhou China department of surgery the First Affiliated Hospital Sun Yat-sen University Guangzhou China department of Gynecology and Obstetrics the First Affiliated Hospital Sun Yat-sen University Guangzhou China department of Obstetrics and Gynecology IVF Program Stanford University Medical Center Palo Alto USA department of Gynecology and Obstetrics Guangdong Provincial People's Hospital Guangzhou China

Aim:To investigate the apoptosis-inducing effect of oridonin,a diterpenoid iso-lated from Rabdosia rubescens,in the human cervical carcinoma HeLa cell ***:A morphological analysis,nuclear condensation,and fragmentation ofchromatin were monitored using Hoechst 33342 *** viability was as-sessed using the 3-(4,5-dimethylthiazol-(2)-yl)-2,5-diphenyl tetrazolium bromide(MTT) *** apoptosis and the apoptosis-related activation in the HeLa cellline were evaluated by flow cytometry and Western ***:Oridoninsuppressed the proliferation of the HeLa cell line in a dose-and *** treatment downregulated the activation of protein kinase B(Akt),the expression of forkhead box class O (FOXO) transcription factor,andglycogen synthase kinase 3 (GSK3).Oridonin also induced the release of cyto-chrome c accompanied by the activation of caspase-3 and poly-adenosine diphos-phate-ribose polymerase *** addition,Z-D(OMe)-E(OMe)-V-D(OMe)-FMK (z-DEVD-fmk),an inhibitor of caspases,prevented caspase-3 activation andabrogated oridonin-induced cell ***,oridonin treatment of the HeLa cellline downregulated the expression of the inhibitor of the apoptosis ***:Our results showed that oridonin-induced apoptosis involved sev-eral molecular *** may suppress constitutively activated targetsof phosphatidylinositol 3-kinase (Akt,FOXO,and GSK3) in the HeLa cell line,inhibiting the proliferation and induction of caspase-dependent apoptosis.

关键词： cervical carcinoma oridonin apoptosis Akt

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PPAR gamma inhibits growth of rat hepatic stellate cells and TGF beta-induced connective tissue growth factor expression

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Acta Pharmacologica Sinica 2006年第6期27卷 715-723页

作者： Kai sun Qian WANG~2 Xiao-hui HUANG department of Hepatobiliary surgery,the first Affiliated Hospital of sun yat-sen university,guangzhou 510080,china department of Hepatobiliary surgery the First Affiliated Hospital of Sun Yat-Sen University Guangzhou China

Aim:To investigate the effect of peroxisome proliferator-activated receptor gamma(PPARγ)activation on the growth of rat hepatic stellate cells(HSC)and trans-forming growth factor beta(TGF-β)-induced connective tissue growth factor(CTGF)***:After being treated with various amounts of thePPARγ natural ligand 15-deoxy-A 12,14-prostaglandin J2(15-d-PGJ2)or syntheticligand GW7845,the status of HSC proliferation and apoptosis were detected byMTT assay and flow ***,HSC were treated with PPARγ-specific antagonist GW9662 prior to the addition of 15-d-PGJ2or GW7845 andwere subsequently stimulated with TGF-β*** mRNA and protein levels ofCTGF expression were detected by semi-quantitative RT-PCR and Western blot-ting *** changes in the HSC were observed by ***:15-d-PGJ2and GW7845 markedly inhibited HSC prolifera-tion and induced cell apoptosis in a dose-dependent ***,PPARγligands significantly suppressed TGF-β1-induced CTGF expression(at both tran-scriptional and post-transcriptional levels)in HSC,and the inhibitory effect wasdramatically,if not completely,abolished by pretreatment with GW9662,sug-gesting that the inhibition was indeed mediated by PPARγ.Moreover,morpho-logical observation revealed that PPARγ activation caused obvious changes ofHSC from activated to quiescent ***:The PPARγligand hasa potent inhibitory effect on the growth of HSC and TGF-β1-induced CTGFexpression,which makes it a potential antifibrotic candidate for the treatment andprevention of hepatic fibrosis.

关键词： PPAR gamma transforming growth factor beta connective tissue growth factor hepatic stellate cells

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Microtubule stability and MAP1B upregulation control neuritogenesis in CAD cells

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Acta Pharmacologica Sinica 2006年第9期27卷 1119-1126页

作者： Wen LI~(2,5)Jin-tang XIA~3 Yue FENG~(4,5)2 Laboratory of surgery,The first Affiliated Hospital,sun yat-sen university,guangzhou 510080,china 3 department of General surgery,The first People's Hospital of guangzhou,guangzhou 510180,china 4 department of Pharmacology,Emory university School of Medicine,USA Laboratory of surgery The First Affiliated Hospital Sun Yat-sen University Guangzhou China department of General surgery The First People's Hospital of Guangzhou Guangzhou China department of Pharmacology Emory University School of Medicine USA

Aim:To study the role of microtubule dynamics and microtubule associated pro-tein 1B(MAP1B)in regulation of the neurite extension in CAD catecholaminergicneuronal cell ***:The neuritogenesis of the CAD cells was abolishedby inhibiting microtubule polymerization with nocodazole and by blocking micro-tubule depolymerization with ***1B and tubulin protein expression wasdetected by Western *** staining of tubulins was observedby fluorescent and confocal ***:Microtubule dynamics wasessential for CAD neurite *** analysis revealed that neurite exten-sion was much more sensitive to nocodazole than to taxol,suggesting a func-tional requirement for highly active microtubule assembly.A remarkableupregulation of MAP IB protein was detected during neurite extension accompa-nied with increased microtubule ***:Upregulation of MAP1Bleads to the stabilization of newly formed microtubules in the developing neurites,which in turn promotes neurite extension.

关键词： microtubule stability microtubule associated protein 1B neuritogenesis

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Addition of tanshinone IIA to UW solution decreases skeletal muscle ischemia-reperfusion injury

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Acta Pharmacologica Sinica 2006年第8期27卷 991-999页

作者： Hong-gang WANG Zhi-yong LI Xiao-lin LIU~2 department of Orthopaedics & Microsurgery,the first Affiliated Hospital of sun yat-sen university,guangzhou 510080,china department of Orthopaedics & Microsurgery the First Affiliated Hospital of Sun Yat-Sen University Guangzhou China

Aim:To investigate whether tanshinone ⅡA could improve the effect of UWsolution for skeletal muscle preservation and to determine the dose range oftanshinone ⅡA providing optimal protection during ischemia and ***:Ischemic rat limbs were perfused with UW solution or UW plus tanshinoneⅡA (UW+T,0.05,0.1,or 0.2 mg/mL) for 0.5h before reperfusion;controls (I/R)received no *** creatine phosphokinase (CPK),aspartate aminotrans-ferase (AST),and lactate dehydrogenase (LDH) were measured pre-ischemia andafter reperfusion (2-h,4-h,and 6-h).Muscle water content,superoxide dismutase(SOD),malondialdehyde (MDA),adenosine triphosphatase (ATPase) were as-sessed pre-reperfusion and after 6-h *** adhesion molecule-1(ICAM-1) and apoptosis were detected after 6-h *** bloodflow was monitored during reperfusion ***:UW and UW+T pre-vented luxury perfusion during reperfusion and inhibited ICAM-1 expression andapoptosis after 6-h *** CPK,AST,and LDH levels in UW ratswere significantly less than those in controls after 2-h reperfusion (no difference,4-h or 6-h reperfusion).After 4-h ischemia,there were significant differences inwater content,MDA,SOD,and ATPase between UW and controls,but no differ-ence after 6-h *** tests with UW+T rats were significantly differentfrom control results at corresponding *** tanshinone doses im-proved ***:UW plus tanshinone ⅡA increased protection againstI/R injury,suggesting that tanshinone ⅡA has clinical value.

关键词： UW solution tanshinone IIA ischemia-reperfusion skeletal muscle

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(-)-Epigallocatechin-3-gallate inhibits growth of gastric cancer by reducing VEGF production and angiogenesis

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World Journal of Gastroenterology 2007年第8期13卷 1162-1169页

作者： Bao-He Zhu, Wen-Hua Zhan, Zheng-Rong Li, Zhao Wang, Yu-Long He, Jun-Sheng Peng, Shi-Rong Cai, Jin-Ping Ma, Chang-Hua Zhang, department of Gastrointestinal & Pancreatic surgery, first Affiliated Hospital, sun yat-sen university Gastric Center of sun yat-sen university, guangzhou 510080, Guangdong Province, china department of Gastrointestinal and Pancreatic surgery First Affiliated Hospital Sun Yat-Sen University Gastric Center of Sun Yat-Sen University Guangzhou 510080 Guangdong Province China.

AIM： To investigate the effect of （-）-epigallocatechin-3-gallate （EGCG） on growth of gastric cancer and its possible mechanism. METHODS： Heterotopic tumors were induced by subcutaneously injection of SGC-7901 cells in nude mice. Tumor growth was measured by calipers in two dimensions. Tumor angiogenesis was determined with tumor microvessel density （MVD） by immunohistology. Vascular endothelial growth factor （VEGF） protein level and activation of signal transducer and activator of transcription 3 （Star3） were examined by Western blotting. VEGF mRNA expression was determined by RT-PCR and VEGF release in tumor culture medium by ELISA. VEGF-induced cell proliferation was studied by MTT assay, cell migration by gelatin modified Boyden chamber （Transwell） and in vitro angiogenesis by endothelial tube formation in Matrigel. RESULTS： Intraperitoneal injection of EGCG inhibited the growth of gastric cancer by 60.4%. MVD in tumor tissues treated with EGCG was markedly reduced. EGCG treatment reduced VEGF protein level in vitro and in vivo. Secretion and mRNA expression of VEGF in tumor cells were also suppressed by EGCG in a dose-dependent manner. This inhibitory effect was associated with reduced activation of Star3, but EGCG treatment did not change the total Star3 expression. EGCG also inhibited VEGF-induced endothelial cell proliferation, migration and tube formation. CONCLUSION： EGCG inhibits the growth of gastric cancer by reducing VEGF production and angiogenesis, and is a promising candidate for anti-angiogenic treatment of gastric cancer.

关键词： Epigallocatechin-3-gallate Angiogenesis Migration Tube formation Vascular endothelial growth factor Signal transducer and activator of transcription 3 Gastric cancer

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Protective mechanism of L-arginine against liver ischemic-reperfusion injury in rats

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Hepatobiliary & Pancreatic Diseases International 2003年第4期2卷 549-552页

作者： Shao-Qiang Li Li-Jian Liang the department of Hepatobiliary surgery, first Affiliated Hospital, sun yat-sen university, guangzhou 510080, china department of Hepatobiliary surgery First Affiliated Hospital Sun Yat-Sen University Guangzhou 510080 China. lesq@***

OBJECTIVE: To investigate the protective effect of L-arginine (L-arg) against liver ischemic-reperfusion (I/R) injury in rat model and its possible mechanism. METHODS: Male Sprague-Dawley rats were randomized into sham group: hepatoduodenal ligament mobilized, but not clamped; control group: hepatic ischemia only; Arg group: 5 minutes before hepatic ischemia, L-arg (200 mg/kg) injected via the dorsal penis vein; Arg+L group: 10 and 5 minutes before hepatic ischemia, L-NAME (30 mg/kg) and L-arg (200 mg/kg) injected via the dorsal penis vein, respectively; and Fink group: 5 minutes before, hepatic ischemia, ZVAD-fmk (15 mg/kg) injected via the dorsal penis vein. The liver was subjected to ischemia for 40 minutes by Pringle's maneuver, and reperfusion was initiated hy removing clamp. The 7-day survival rate, alanine transaminase (ALT) level, caspase-3 activity and apoptotic hepatocyte count were compared among these groups. RESULTS: After 40 minutes of ischemia and 6 hours of reperfusion, the 7-day survival rate of the Arg group was significantly higher than that of the control and Arg+L groups (P<0.05). The ALT Level, caspase-3 activity and apoptotic hepatocyte count in the Arg group were decreased significantly than those in the control and Arg+L groups (P<0. 01). The caspase-3 activity and apoptotic hepatocyte count in the Arg group were slightly higher than those in the sham and Fmk groups, but there was no statistical significance. CONCLUSIONS: L-arg could ameliorate liver I/R injury and the possible protective mechanism is inhibition of hepatocyte apoptosis via inhibition of caspase-3 activity by nitric oxide synthesis.

关键词： liver ischemia nitirc oxide caspase-3 rat

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Interferon-α enhances sensitivity of human osteosarcoma U2OS cells to doxorubicin by p53-dependent apoptosis

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Acta Pharmacologica Sinica 2007年第11期28卷 1835-1841页

作者： Xiang-wei YUAN~2,Xiao-feng ZHU~(3,5),Xiu-fang HUANG~4,Pu-yi SHENG~2,Ai-shan HE~2,Zi-bo YANG~2,Rong DENG~3,Gong-kanFENG~3,Wei-ming LIAO~(2,5)~2 department of Orthopedic surgery,The first Affiliated Hospital,sun yat-sen university,guangzhou 510080,china ~3 State Key Laboratoryof Oncology in South china,Cancer Center,sun yat-sen university,guangzhou 510060,china ~4 department of Pathology,The AffiliatedJiangmen Hospital,sun yat-sen university,Jiangmen 529030,china department of Orthopedic surgery The First Affiliated Hospital Sun Yat-sen University Guangzhou China State Key Laboratory of Oncology in South china Cancer Center Sun Yat-sen University Guangzhou China department of Pathology The Affiliated Jiangmen Hospital Sun Yat-sen University Jiangmen China

Aim:To determine whether interferon-α(IFNα) can enhance doxorubicinsensitivity in osteosarcoma cells and its molecular ***:Cellviability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide *** was studied using Flow cytometry analysis,Hoechst33258 staining,DNA fragmentation assay,as well as the activation ofcaspase-3 and poly (ADP-ribose) *** expression was detectedby Western *** dependence of p53 was determined using ***:IFNα increased doxorubicin-induced cytotoxicity to a muchgreater degree through apoptosis in human osteosarcoma p53-wild U2OS cells,but not p53-mutant MG63 ***α markedly upregulated p53,Bax,Mdm2,andp21,downregulated Bcl-2,and activated caspase-3 and PARP cleavage in re-sponse to doxorubicin in U2OS ***,the siRNA-mediated silencing ofp53 significantly reduced the IFNα/doxorubicin combination-induced cytotoxic-ity and PARP ***:IFNα enhances the sensitivity of humanosteosarcoma U2OS cells to doxorubicin by p53-dependent *** propercombination with IFNα and conventional chemotherapeutic agents may be a ra-tional strategy for improving the treatment of osteosarcoma with functional p53.

关键词： Interferon-α doxorubicin apoptosis p53 osteosarcoma

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Establishment of multiplexed,microsphere-based flow cytometric assay for multiple human tumor markers

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Acta Pharmacologica Sinica 2007年第12期28卷 2011-2018页

作者： Kai sun,Qian WANG~2,Xiao-hui HUANG,Mao-chuan ZHEN,Wen LI,Long-juan ZHANGdepartment of Hepatobiliary surgery,the first Affiliated Hospital of sun yat-sen university,guangzhou 510080,china department of Hepatobiliary surgery the First Affiliated Hospital of Sun Yat-Sen University Guangzhou China

Aim:The multiplexed,microsphere-based flow cytometric assay (MFCA) for mul-tiple human tumor markers was established for the early screening and detectionof suspected cancer ***:Covalent coupling of capture antibodiesdirected against their respective tumor markers to fluorescent microspheres wasperformed by following the protocols recommended by a commercial corporationwith some *** coupling efficiency and cross-reactivity were iden-tified by the Luminex 100 system and associated *** standard curvewas constructed by using serial dilution of recombinant tumor marker standardsand was validated by comparison with ELISA for quantifying the tumor markers inserum ***:The identifications revealed that the coupling proce-dures were successful without non-specific cross-reactivity and the standardcurve was highly ***,it was necessary to ensure the quality con-trol of the coupling process since slight variations in the coupling procedurescould profoundly affect the density of capture reagents coupled to themicrospheres and consequently adversely affect the assay *** additionto its multi-analyte capability,the MFCA system had definite advantages,such ashigher reproducibility,greater dynamic range of measurement,and considerablyless preparation time and labor over the conventional"gold standard",which wasthe ***:The successful establishment of the MFCA system forthe simultaneous detection of multiple tumor markers will provide the foundationfor the further study of clinical applications.

关键词： fluorescent microsphere microsphere-based flow cytometric assay tumor marker

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Temporal Control of Cre Recombinase-mediated in Vitro DNA Recombination by Tet-on Gene Expression System

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 133-138页

作者： Zhong-Min GUO Kang XU Ying YUE Bing HUANG Xin-Yan DENG Nu-Qi ZHONG Xun HONG Xi-Gu CHEN Dong XIAO Center of Experimental Animals.sun yat-sen (Zhongshan) university.guangzhou 510080.china The first Affiliated Hospital of sun yat-sen university.sun yat-sen university.Guang-hou 510080.china Center of Experimental Animals Sun Yat-Sen (Zhongshan) University Guangzhou 510080 China The first Affiliated Hospital of sun yat-sen university Sun Yat-Sen University Guangzhou 510080 China

Conditional gene expression and gene deletion are important experimental approaches forexamining the functions of particular gene products in mouse models,These strategies exploiting Cre-medi-ated site-specific DNA recombination have been incorporated into transgenic and gene-targeting proceduresto allow in vivo manipulation of DNA in embryonic stem cells(ES cells)or living *** Cre/lox Psystem has become widely used in conditional gene targeting,conditional gene repair and activation,induc-ible chromosome translocation,and chromosome *** this project,we have employed the univer-sal transgenic system and the liver-specific promoter system for tightly temporal and liver-specific control ofCre gene expression in mice that(1)integrates the advantages of the Tet-on gene expression system and Cre//ox P site-mediated gene activation,and(2)simplifies the scheme of animal crosses through a combination oftwo control elements in a single transgene.A liver-specific apoE promoter was inserted into the promotercloning site upstream of the rtTA cassette of pCore construct to generate the transgene construct pApoErtTA-tetO-Cre,followed by demonstrating stringent regulation of doxycycline(Dox)-induced Cre-mediated re-combination in the lox P-flanked transcription STOP cassette-modified BEL-7402 *** is to say,in theabsence of Dox,the Cre gene is not expressed and will not induce site-specific recombination between twolox P sites,whereas on exposure to Dox,the Cre gene will be expressed and the recombination will ***,these data indicate that the Tet-on gene expression system is able to successfully and stringentlycontrol Cre expression in vitro,which lays a solid foundation for efficient and spatio-temporal Cre geneactivation in transgenic mice.

关键词： conditional gene regulation Cre recombinase human apolipoprotein E gene(apoE)promoter Tet-on gene expression system universal transgenic system

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Protective effect of membrane cofactor protein against complementdependent injury

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Acta Pharmacologica Sinica 2005年第8期26卷 987-991页

作者： Dong XU, Shou-jian HUANG~1, Jin-qun WANG, Chu-kun WU department of Pharmacology, sun yat-sen Medical College of sun yat-sen university, guangzhou 510080, china department of Pharmacology Sun Yat-Sen Medical College of Sun Yat-Sen University Guangzhou China

Aim: To evaluate the protective role of membrane cofactor protein (MCP CD46) on complement-dependent injury. Methods: MCP was separated by ion exchange chromatography on a DEAE sephadex A-50 column from pig erythrocyte ghosts. Its protective effect was tested in models such as cobra venom factor (CVF)-induced platelet metamorphosis and aggregation, human serum-induced injury in isolated working guinea pig heart and reverse passive Arthus reaction. Results: MCP inhibited CVF-induced platelet metamorphosis with an IC50of 56.7 mg/L±2.6 mg/L, and prevented injury induced by activated complement in isolated working guinea pig hearts. In the rat model of reverse Arthus reaction, MCP relieved the skin lesions induced by immune complexes. Conclusion: MCP has a protective effect against complement-dependent injury.

关键词： CD46 antigen complement heart reverse Arthus reaction

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