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Effect of curcumin on multidrug resistance in resistant human gastric carcinoma cell line SGC7901/VCR

Acta Pharmacologica Sinica 2005年第8期26卷 1009-1016页

作者： Xiao-qing TANG~1, Hu BI~1, Jian-qiang FENG~2, Jian-guo CAO~3 ~1 department of physiology, Nanhua university, Hengyang 421001, china ~2 department of physiology, zhongshan medical college, sun yat sen university, guangzhou 510080, china ~3 Institute of Oncology, Nanhua university, Hengyang 421001, china department of physiology Nanhua University Hengyang China department of physiology Zhongshan Medical College Sun Yatsen University Guangzhou China Institute of Oncology Nanhua University Hengyang China

Aim: To investigate the reversal effects of curcumin on multidrug resistance （MDR） in a resistant human gastric carcinoma cell line. Methods: The cytotoxic effect of vincristine （VCR） was evaluated by MTT assay. The cell apoptosis induced by VCR was determined by propidium iodide （PI）-stained flow cytometry （FCM） and a morphological assay using acridine orange （AO）/ethidium bromide （EB） dual staining. P-glycoprotein （P-gp） function was demonstrated by the accumulation and efflux of rhodamine 123 （Rh123） using FCM. The expression of P-gp and the activation of caspase-3 were measured by FCM using fluorescein isothiocyanate （FITC）-conjugated anti-P-gp and anti-cleaved caspase-3 antibodies, respectively. Results: Curcumin, at concentrations of 5 μmol/L, 10 μmol/L, or 20 μmol/L, had no cytotoxic effect on a parent human gastric carcinoma cell line （SGC7901） or its VCR-resistant variant cell line （SGC7901/VCR）. The VCR-IC50value of the SGC7901/VCR cells was 45 times more than that of the SGC7901ceUs and the SGC7901/VCR cells showed apoptotic resistance to VCR. SGC7901/VCR cells treated with 5 μmol/L, 10μmol/L, or 20 μmol/L curcumin decreased the IC50value of VCR and promoted VCR-mediated apoptosis in a dose-dependent manner. Curcumin （10 μmol/L） increased Rh123 accumulation and inhibited the effiux of Rh123 in SGC7901/VCR cells, but did not change the accumulation and efflux of Rh123 in SGC7901 ceils. P-gp was overexpressed in SGC7901/VCR cells, whereas it was downregulated after a 24-h treatment with curcumin （10 μmol/L）. Resistant cells treated with 1 μmol/L VCR alone showed 77% lower levels of caspase-3 activation relative to SGC7901 cells, but the activation of caspase-3 in the resistant cell line increased by 44% when cells were treated with VCR in combination with curcumin. Conclusion: Curcumin can reverse the MDR of the human gastric carcinoma SGC7901/VCR cell line. This might be associated with decreased P-gp function and expression, and the promotion of caspase-3

关键词： curcumin multidrug resistance apoptosis P-glycoprotein caspase-3

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Proteomics-based Identification of Proteins with Altered Expression Induced by 12-O-Tetradecanoylphorbol 13-acetate In Nasopharyngeal Carcinoma CNE2 Cells

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Acta Biochimica et Biophysica Sinica 2005年第2期37卷 97-106页

作者： Pei-Zhou JIANG Ming GAN Hua HUANG Xin-Ming SHEN Shuang WANG Kai-Tai YAO Cancer Research Institute.Southern medical university,guangzhou 510515,china department of Histology and Embryology.Southern medical university,guangzhou 510515,china department of Parasitology,zhongshan medical college.sun yat-sen university,guangzhou 510080.china Cancer Research Institute Southern Medical UniversityGuangzhou 510515 China department of Histology and Embryology Southern Medical UniversityGuangzhou 510515 China department of Parasitology Zhongshan Medical College SUN Yat-sen UniversityGuangzhou 510080 China

Nasopharyngeal carcinoma(NPC)is a malignancy with high incidence in Southern china andSouth-East *** studies indicate that chemical carcinogen promoters,such as 12-O-tetradecanoylphorbol-13-acetate(TPA),are important ... 详细信息

关键词： 12-O-tetradecanoyl-phorbol-13-acetate nasopharyngeal carcinoma proteomics apoptosis

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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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Temporal and Tight Hepatitis C Virus Gene Activation in Cultured Human Hepatoma Cells Mediated by a Cell-Permeable Cre Recombinase

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Acta Biochimica et Biophysica Sinica 2004年第10期36卷 687-694页

作者： Dong XIAO*, Kang XU1, Ying YUE, Zhong-Min GUO, Bing HUANG, Xin-Yan DENG, Huan TANG2, and Xi-Gu CHEN* Center of Experimental Animals, zhongshan university, guangzhou 510080, china 1The first affiliated hospital of sun yat-sen university, zhongshan university, guangzhou 510080, china 2The Third Military medical university, Chongqing 400010, china Center of Experimental Animals Zhongshan UniversityGuangzhou 510080 China The first affiliated hospital of sun yat-sen university Zhongshan UniversityGuangzhou 510080 China The Third Military medical universityChongqing 400010 China

Conditional gene expression has greatly facilitated the examination of the functions of particular gene products. Using the Cre/lox P switching expression system, we plan to develop efficient conditional transgeneactivation of hepatitis C virus core protein (HCV-C) cDNA (nucleotide 342–914) in the transgenic mice to overcome “immune tolerance” formed during the embryonic period and “immune escape” against hepatitis virus antigen in our project. To use this system in vivo, the dormant transgenic construct, i.e., pApoE-SCS-EGFP-HCV-C, was generated using techniques of standard molecular biology. The liver- specific human apoE promoter was here used to target expression of genes of interest (EGFP and HCV-C) to murine liver. Prior to generating the transgenic mice, the availability of Cre/lox P system and construct functionality were successfully verified by a cell-free recombination system and via checking the expression of EGFP and HCV-C in the human hepatoma cells at the mRNA and protein levels. These results suggest that the Cre/lox P system could tightly control expression of EGFP and HCV-C in vitro, which laid a solid foundation to conditionally activate expression of target gene(s) in transgenic mice by Cre-mediated site-specific recombination.

关键词： hepatitis C virus Cre/lox P switching expression system cell-free and intracellular recombination systems cell-permeable Cre recombinase EGFP human hepatoma cells

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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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Activation of PI3-K/Akt pathway for thermal preconditioning to protect cultured cerebellar granule neurons against low potassium-induced apoptosis

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Acta Pharmacologica Sinica 2007年第2期28卷 173-179页

作者： Lin CAO~(2,3) De-xiong CAO~3 Xing-wen SU~2 Li-jun CHEN~2 Ai-lin LIU~2 Wei-jian JIAN~2 Yu-ping WU~2 Peng-xin QIU~2 Guang mei YAN~(2,4)2 department of Pharmacology,zhongshan medical School,sun yat-sen university,guangzhou 510080,china 3 department of Anesthesiology The Second Affiliated Hospital,sun yat-sen university,guangzhou 510120,china department of Pharmacology Zhongshan Medical School Sun Yat-sen University Guangzhou China department of Anesthesiology The Second Affiliated Hospital Sun Yat-sen University Guangzhou China

Aim:This study was designed to investigate whether the activation of thephosphatidylinositol 3-kinase(PI3-K)/Akt pathway is required for thermal pre-conditioning to protect rat cerebellar granule neurons(CGN)against apoptosisinduced by low potassium,and to explore the possibility of a link between theupregulated heat shock protein(HSP)70 expression and Akt activation in theacquisition of neuroprotection induced by thermal ***:CGN cultured for 8 d in vitro were switched to 5K medium for 24 h after thermalpreconditioning(TP;43.5 ℃ for 90 min,then 37 ℃ for 1 h).To study the role of thePI3-K/Akt pathway,a PI3-K inhibitor,LY294002(20 μmol/L)was added into thecultures 1 h before TP.3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bro-mide(MTT)assay and fluorescein diacetate staining were used to determine *** 33258 staining and agar gel electrophoresis were used to testthe morphological and biological characters of *** blot analysis wasemployed to detect the levels of phospho-Akt,phospho-glycogen synthase ki-nase 3β(GSK3β) Akt,GSK3β,and ***:TP protected CGN againstapoptosis induced by low ***294002 inhibited the neuroprotectiveeffect on CGN induced by *** induced a robust activation of Akt and theinactivation of GSK3β via ***,the activation of the PI3-K/Aktpathway by TP persisted for 24 h in the 5K ***294002(20 μmol/L)failedto inhibit the upregulated HSP70 expression induced by ***:Theactivation of the PI3-K/Akt pathway is required for TP to protect CGN againstapoptosis induced by low potassium,but the neuroprotective effect by Akt acti-vation is not mediated through the downstream induction of HSP70 expression.

关键词： thermal preconditioning heat cerebellum neurons apoptosis LY294002 phospho Akt phospho-GSK3β HSP70

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Purification and partial characterizations of coagulant protein Fla from Daboia russelli siamensis (Myanmar) venom

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Acta Pharmacologica Sinica 2007年第10期28卷 1580-1584页

作者： Huan-huan sun~(1,3),Qi CHEN~(2,3),Xi LIN~1,Jia-shu CHEN~(1,4),Peng-xin QIU~1,Guang-mei YAN~~1 department of Pharmacology,zhongshan medical college,sun yat-sen university guangzhou 510080,china ~2 Guangdong Institute forDrug Control,guangzhou 510000,china department of Pharmacology Zhongshan Medical College Sun Yat-Sen University Guangzhou China Guangdong Institute for Drug Control Guangzhou China

Aim:To purify and characterize the coagulant protein FIa from Daboia russellisiamensis (Myanmar) ***:FIa was purified from Daboia russellisiamensis (Myanmar) venom by ion-exchange chromatography on CM-SephadexC-50,and gel filtration on Sephadex G-75 and a Superdex 75 *** hemo-static activity of FIa was determined by the method of Williams and *** chromogenic substrates were used respectively to determine the activa-tion of factor X and *** fibrinogen-clotting activity of FIa wasdetermined by the method of Gao et *** saline was used as a negativecontrol while factor Xa and thrombin were used as positive controls,***:FIa,a coagulant protein,was achieved by ion-exchange chromatogra-phy and gel filtration with a molecular weight of 34 479 and an isoelectric point *** was shown to have strong hemostatic *** hemostatic activity of0.5 mg FIa was equal to that of 1.5625 u *** primarily activated factor X,however,had no influence on prothrombin,nor did it cleave or clot ***:FIa is a factor X-activating enzyme,which could activate factor X tofactor Xa,but has no effect on prothrombin and fibrinogen.

关键词： snake venoms Daboia russelli siamensis factor X blood coagulation

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

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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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Non-invasive Prenatal Gene Diagnosis: Progress through Cell-free Fetal DNA and RNA in Maternal Plasma and Urine

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中山大学学报（医学科学版） 2008年第S2期29卷 140-142页

作者： GUO Xun-yang, GUO Yi-bin ( department of medical Genetics, zhongshan School of Medicine, sun yat-sen university, guangzhou 510080, china )

Non-invasive prenatal gene diagnosis has been developed rapidly in the recent years, and numerous medical researchers are focusing on it. Such techniques could not only achieve prenatal diagnosis accurately, but also prevent tangential illness in fetuses and thus, reduce the incidence of diseases. Moreover, it is non-invasive prenatal gene diagnosis that prevents potential threaten and danger to both mothers and fetuses. Therefore, it is welcomed by clinical gynecologist and obstetrian, researchers of medical genetics, and especially, pregnancies. This review article touches briefly on the advanced development of using cell-free DNA, RNA in maternal plasma and urine for non-invasive prenatal gene diagnosis.

关键词： non-invasive prenatal gene diagnosis cell-free fetal DNA and RNA DNA and RNA detection maternal urine maternal plasma

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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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