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Hypaconitine inhibits TGF-β1-induced epithelial–mesenchymal transition and suppresses adhesion,migration,and invasion of lung cancer A549 cells

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Chinese Journal of Natural Medicines 2017年第6期15卷 427-435页

作者： FENG Hai-Tao ZHAO Wen-Wen lu jin-jian WANG Yi-Tao CHEN Xiu-Ping State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical Sciences University of MacaoMacao China

Epithelial-mesenchymal transition(EMT) has been implicated in tumor invasion and metastasis and provides novel strategies for cancer therapy. Hypaconitine(HpA), a diester-diterpenoid alkaloid isolated from the root of the Aconitum species, exhibits anti-inflammatory, analgesic, and especially, cardiotoxic activities. Here, we reported the anti-metastatic potentials of HpA in transforming growth factor-β1(TGF-β1)-induced EMT in lung cancer A549 cells. The cytotoxic effect of HpA was determined by MTT assay. A549 cells were treated with TGF-β1 with or without HpA co-treatment, and the morphological alterations were observed with a microscopy. The expression of E-cadherin, N-cadherin, and NF-κB was determined by both Western blotting and immunofluorescence analyses. The adhesion, migration, and invasion were detected with Matrigel, wound-healing, and transwell assays, respectively. The expression of Snail was determined by Western blotting. The expression of NF-κB p65, IκBα, and p-IκBα in nuclear and cytosolic extracts was assessed by Western blotting. The results showed that low concentration of HpA(lusion, these results showed that HpA inhibited TGF-β1-induced EMT in A549 cells, which was possibly mediated by the inactivation of the NF-κB signaling pathway, providing an evidence for anti-cancer effect of HpA.

关键词： EMT Hypaconitine TGF-β1 NF-κB

Cucurbitacin B suppresses metastasis mediated by reactive oxygen species(ROS) via focal adhesion kinase(FAK) in breast cancer MDA-MB-231 cells

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Chinese Journal of Natural Medicines 2018年第1期16卷 10-19页

作者： luO Wei-Wei ZHAO Wen-Wen lu jin-jian WANG Yi-Tao CHEN Xiu-Ping State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical Science University of Macao

Metastasis is responsible for the majority of cancer-related deaths and prevention of metastasis remains a big challenge for cancer therapy. Cucurbitacin B(Cuc B) is a natural triterpenoid with potent anticancer activities while its effect on metastasis remains unclear. In the present study, the inhibitory effect and mechanisms of Cuc B on metastasis were investigated in MDA-MB-231 breast cancer cells. The cells were treated with or without Cuc B, and the cytotoxicity was determined by MTT assay. The effect of Cuc B on metastasis was evaluated with wound healing, transwell, and adhesion assays. Furthermore, the adhesion of cancer cells to endothelial cells was determined. The protein expression was determined by Western blotting. Cuc B(lular reactive oxygen species(ROS) generation, which could be reduced by N-acetyl-l-cysteine(NAC). In addition, NAC pretreatment could reverse Cuc B-induced suppression of migration and adhesion, expression of FAK, but showed no effect on paxillin expression. In summary, Cuc B suppressed ROS-dependent metastasis through FAK pathway in breast cancer MDA-MB-231 cells, demonstrating novel mechanisms for the anticancer effects of Cuc B.

关键词： Cucurbitacin B Metastasis ROS FAK Breast Cancer

Proteomic analysis of hepatocellular carcinoma HepG2 cells treated with platycodin D

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Chinese Journal of Natural Medicines 2015年第9期13卷 673-679页

作者： lu jin-jian lu De-Zhao CHEN Yu-Fei DONG Ya-Ting ZHANG Jun-Ren LI Ting TANG Zheng-Hai YANG Zhen State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical Sciences University of Macao College of Life Sciences Zhejiang Chinese Medical University

Platycodin D(PD), a triterpenoid saponin isolated from Platycodonis Radix, is a famous Chinese herbal medicine that has been shown to have anti-proliferative effects in several cancer cell lines. The aim of this study was to determine the changes in cellular proteins after the treatment of hepatocellular carcinoma HepG2 cells with PD using proteomics approaches. The cell viability was determined using the MTT assay. The proteome was analyzed by two-dimensional difference gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blot analysis was used to confirm the expression of changed proteins. Our results showed that PD inhibited the proliferation of HepG2 cells in concentration- and time-dependent manners. Sixteen proteins were identified to be up-regulated in PD-treated HepG2 cells, including ATP5 H, OXCT1, KRT9, CCDC40, ERP29, RCN1, ZNF175, HNRNPH1, HSP27, PA2G4, PHB, BANF1, TPM3, ECH1, LGALS1, and MYL6. Three proteins(i.e., RPS12, EMG1, and KRT1) decreased in HepG2 cells after treatment with PD. The changes in HSP27 and PHB were further confirmed by Western blotting. In conclusion, our results shed new lights on the mechanisms of action for the anti-cancer activity of PD.

关键词：药学药剂学调剂学剂型

Towards a better understanding of the therapeutic potential of Rhizoma Alismatis: A phytochemical and pharmacological review

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中国药理学与毒理学杂志 2016年第10期30卷 1083-1084页

作者： ZHANG Le-le WANG Yi-tao lu jin-jian State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of Macao

Rhizoma Alismatis(RA),the dried rhizome of Alisma orientalis(Sam.)Juzep,isa widely used traditional herbal medicine popularly known as″Ze Xie″in Chinese.RA is an important crude herb component of several famous Chinese preparations including Zexie decoction,Liu Wei Di Huang pills,Wu Ling power and so on.It has been used to treat various ailments such as dysuria,edema,nephropathy,hypertension,hyperlipidemia and diabetes,etc.A wide range of chemical compounds mainly triterpenes,diterpenes and sesquiterpenes have been isolated from RA,among which the protostane-type triterpenes termed alisols attract the most attention owing to their unique chemical structure and various biological activity.The extract and active compounds of RAhave been demonstrated to possess a wide spectrum of pharmacological effects which support its clinical use,e.***.diuretic,hypoglycemic,hypolipidemic,hepatoprotective,preventing bone loss,anti-inflammatory andantivirus activities.In our previous studies,we have investigatedthe anti-proliferative activities of terpenoids isolated from RA and their structure-activity relationships.Alisol B 23-acetate was demonstrated to induce G1 phase cell cycle arrest and apoptosis,and inhibit migration and invasion of ovarian cancer cells.The present paper reviews the up-to-date information concerning the ethnopharmacology,phytochemistry,pharmacology and toxicology of RA.This information will be helpful for a better understanding of therapeutic potential of RA as wel as the further explorations.

关键词： Rhizoma Alismatis ethnopharmacology phytochemistry pharmacology

Protection of Total Flavonoid Fraction from Nervilia fordii on Lipopolysaccharide-induced Acute lung Injury in Rats

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Chinese Herbal Medicines 2012年第4期4卷 287-293页

作者： HUANG Ming-qing XIE You-liang LAI Xiao-ping LIN Ling XU Yin-ji lu jin-jian CHEN Xiu-ping College of Pharmacy Fujian University of Traditional Chinese MedicineFuzhou 350108China Institute of Drug Research and Development Guangzhou University of Chinese MedicineGuangzhou 510405China Guangdong Provincial Hospital of Traditiona lChinese Medicine Guangzhou 510120China State Key Laboratory of Quality Rese arch in Chinese Medicine University of MacaoMacao 999078China

Objective To investigate the effects of total flavonoid fraction(TFF) from Nervilia fordii on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in rats,and to explore their protective mechanism.Methods LPS-induced ALI model was established by LPS(5 mg/kg) injection via left cervical vein.Blood samples were collected from the cervical artery of all rats at 5 and 6 h after LPS challenge for arterial blood gas test and cytokines measurements,and pulmonary microvascular permeability(PMP),lung wet/dry weight ratio(W/D),and pathological features were observed.Results Phytochemical study showed that the TFF contained 67.3% of flavonoids expressed in rutin and three flavone glycosides.The TFF pretreatment(6.24 and 12.48 mg/kg) attenuated the partial arterial pressure of oxygen decline in blood significantly,and decreased the PMP and lung W/D in ALI rats.In addition,the TFF(6.24 and 12.48 mg/kg) also ameliorated the LPS-induced lung damages including alveolar edema,neutrophils infiltration,alveolar hemorrhage,and thickening of the alveolar wall.Furthermore,the treatment with the TFF(6.24 and 12.48 mg/kg) also down-regulated the levels of pro-inflammatory cytokines,such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and intercellular adhesion molecule-1(ICAM-1),and up-regulated the level of anti-inflammatory cytokine IL-10 in serum of ALI rats simultaneously.Conclusion These results suggest that the TFF could protect LPS-induced ALI in rats,which may be mediated,at least in part,by adjusting the production of inflammatory cytokines including TNF-α,IL-6,ICAM-1,and IL-10.

关键词： acute lung injury cytokine Nervilia fordii pulmonary microvascular permeability total flavonoid fraction

Nagilactone E increases PD-L1 expression through activation of c-Jun in lung cancer cells

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Chinese Journal of Natural Medicines 2020年第7期18卷 517-525页

作者： CHEN Yu-Chi HUANG Mu-Yang ZHANG Le-Le FENG Zhe-Ling jianG Xiao-Ming YUAN luo-Wei HUANG Run-Yue LIU Bo YU Hua WANG Yi-Tao CHEN Xiu-Ping LIN Li-Gen lu jin-jian State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of MacaoMacao 999078China School of Medicine Chengdu UniversityChengdu 610106China The Second Affiliated Hospital of Guangzhou University of Chinese Medicine(Guangdong Provincial Hospital of Chinese Medicine) Guangzhou 510120China

Nagilactone E(NLE),a natural product with anticancer activities,is isolated from Podocarpus nagi.In this study,we reported that NLE increased programmed death ligand 1(PD-L1)expressions at both protein and mRNA levels in human lung cancer cells,and enhanced its localization on the cell membrane.Mechanistically,NLE increased the phosphorylation and expression of cJun,and promoted the localization of c-Jun in the nucleus,while silencing of c-Jun by small interfering RNA(siRNA)reduced NLEinduced PD-L1.Further study showed that NLE activated the c-Jun N-terminal kinases(JNK),the upstream of c-Jun,and its inhibitor SP600125 reversed the NLE-increased PD-L1.Moreover,NLE-induced PD-L1 increased the binding intensity of PD-1 on the cell surface.In summary,NLE upregulates the expression of PD-L1 in lung cancer cells through the activation of JNK-c-Jun axis,which has the potential to combine with the PD-1/PD-L1 antibody therapies in lung cancer.

关键词： Programmed death ligand 1 Nagilactone E lung cancer c-Jun JNK

Effects of garcinone E on ovarian cancer cells:endoplasmic reticulum stress,apoptosis,migration and invasion inhibition

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中国药理学与毒理学杂志 2016年第10期30卷 1075-1076页

作者： XU Xiao-huang LI Ting ZHANG Le-le CHEN Xin GUO Xia CHEN Xiu-ping WANG Yi-tao LIN Li-gen lu jin-jian State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of Macao

OBJECTIVE Garcinone E,a xanthone derivative isolated from mangosteen,has been reported to have cytotoxic capacities.We aim to study the anticancer effects of garcinone E on ovarian cancer cells.METHODS MTT assay,LDH release assay,Hoechst33342 staining,AnnexinⅤ/PI staining,and JC-1 staining were used to test the antitumor abilities on several ovarian cancer cell lines,and wound-healing assay,transwell assay,and gelatin zymography were carried out to examine the migratory and invasive effects of malignant ovarian cancer HEY cells.Relevant m RNA transcription and protein expression were evaluated by Western blotting.RESULTS Garcinone E treatment significantly inhibited the proliferation and caused cell death on ovarian cancer HEY,A2780 as well as A2780-palicitaxel resistant cells.It induced endoplasmic reticulum stress in HEY cells and activated IRE signaling pathway,which provided protection on HEY cells.Meanwhile,knocking down of IRE-1αby si RNA further activated caspase cascade and caused more cell death.On the other hand,garcinone E eliminated migrative ability of HEY cells by reducing the expression of Rho A and Rac,and it blockaded invasion by down-regulating the protein level of MMP-9 and-2,and up-regulating the protein level of TIMP-1 and-2.Garcinone E-reduced activities of MMP-9 and-2 were confirmed by gelatin zymography assay.CONCluSION Our finding demonstrated that garcinone E exerts anticancer activities by inducing apoptosis,and suppressing migration and invasion on ovarian cancer cells,which demonstrated certain therapeutic potential of garcinone E on ovarian cancer.

关键词： ovarian cancer garcinone E apoptosis metastasis ER stress

Suppression of AOM/DSS-induced colorectal cancer by scutellarin through downregulation of Wnt signaling pathway activity

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中国药理学与毒理学杂志 2021年第10期35卷 739-740页

作者： ZENG Sha ZHAO Hui CHEN Li SUN Qiang REN Shan LIU Mao-lun YANG Han TANG Shun lu jin-jian XU Hai-bo State Key Laboratory of Southwestern Chinese Medicine Resources Department of PharmacologyChengdu University of Traditional Chinese MedicineChengdu 611137China Institute of Chinese Medical Sciences University of MacaoMacaoChina

OBJECTIVE To investigate the therapeutic effect of scutellarin on colitis-associated cancer(CAC)and its underlying mechanism based on Wnt/β-catenin signaling pathway.METHODS The mouse model of CAC was established by azomethane oxide(AOM)and sodium dextran sulfate(DSS),followed by scutellarin treatment,with recording the body weight,diarrhea and hematochezia.After sacrificing the mice,the colorectal length and colorectal tumor were assessed.The levels of pro-inflammatory factors TNF-αand IL-6 in mice′s sera were measured by the enzyme-linked immunosorbent assay(ELISA).The colorectal lesions were appraised by hematoxylin and eosin(H&E)staining.Theβ-catenin level in CAC tissues was probed by immunofluorescent analysis.The apoptosis-related genes Bax and Bcl-2,and Wnt signaling pathway-related genesβ-catenin,GSK-3β,TCF4,c-Myc and cyclin D1 were detected by real-time quantitative RT-PCR(RT-qPCR).Finally,Western blotting analysis(WB)was employed to examine the expressions of the apoptosis and Wnt signaling pathway-related proteins.RESULTS Scutellarin significantly improved AOM/DSS-caused weight loss,colorectal length shortening,and tumor growth in mice(P<0.01).Meanwhile,colorectal lesions could be substantially alleviated by scutellarin.ELISA results showed that the levels of pro-inflammatory factors TNF-αand IL-6 were drastically lessened(P<0.01).Scutellarin also sharply inhibited the nuclear translocation ofβ-catenin,as evidenced by the reduction in the nuclear level ofβ-catenin protein.In addition,scutellarin attenuated the mRNA expression of Wnt signaling pathway-relatedβ-catenin,TCF4,c-Myc and cyclin D1,whereas it heightened GSK-3βmRNA level.These results were consolidated by WB analysis,which indicated that scutellarin could mitigate the protein levels of phospho-GSK-3β,β-catenin,TCF4,c-Myc and cyclin D1,with the increase in GSK-3βprotein in CAC tissue.Moreover,scutellarin could induce the apoptosis of CAC,demonstrated by enhanced expression of Bax and diminished expression of Bcl-2 in both mRNA and protein level

关键词： scutellarin colorectal cancer Wnt signaling pathway AOM/DSS

Inhibition of colorectal cancer by ursolic acid via noncanonical Hedgehog signaling pathway

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中国药理学与毒理学杂志 2021年第10期35卷 759-760页

作者： CHEN Li SUN Qiang ZENG Sha ZHAO Hui LIU Mao-lun YANG Han REN Shan MING Tian-qi lu jin-jian XU Hai-bo State Key Laboratory of Southwestern Chinese Medicine Resources Department of PharmacologyChengdu University of Traditional Chinese MedicineChengdu 611137China Institute of Chinese Medical Sciences University of MacaoMacaoChina

OBJECTIVE To identify the inhibitory effect of ursolic acid on the colorectal cancer HCT116 cells in vitro and in vivo,and to explore the underlying mechanism.METHODS The smoothened(SMO)gene-silenced human colorectal cancer HCT116^(hSMO)cell line was established by transfection with the lentivirus carrying SMO shRNA.The cytotoxic effect of ursolic acid on HCT116^(hSMO)cells was determined by MTT assay.The effect of ursolic acid on the migration of HCT116^(hSMO)cells was studied by wound healing assay.The effect of ursolic acid on apoptosis of HCT116^(hSMO)cells was explored by Hoechst33342/PI double staining and flow cytometry.The effects of ursolic acid on the expressions of apoptotic marker gene Bcl-2,Bax,caspase-3 and caspase-9 were measured by real-time quantitative RT-PCR(RT-qPCR)and Western blotting(WB)analysis.RT-qPCR and WB were used to examine the relationship between GLI1,c-Myc expression and PI3K/Akt pathway to further investigate the mechanism of GLI1 activation in HCT116^(hSMO)cells.The effects of ursolic acid on the expressions of GLI1,p-Akt,Akt,c-Myc,SHH and SUFU of noncanonical Hedgehog pathway were evaluated by RT-qPCR and WB assays.Xenograft nude mouse model bearing HCT116^(hSMO)cells was established and intraperitoneally treated with ursolic acid to investigate the effect on tumor growth in vivo.The body weight and tumor size of mice were assessed regularly every 2 d.The effect of ursolic acid on the apoptosis of tumor tissue was determined by TUNEL assay.The expressions of Bcl-2,Bax,GLI1,p-Akt,Akt,c-Myc,SHH,SUFU mRNA and proteins were measured by RT-qPCR and WB.The levels of Bcl-2,Bax,GLI1,p-Akt,c-Myc and SHH proteins in tumor tissues were also evaluated by immunohistochemistry.RESULTS Ursolic acid significantly inhibited the growth and migration of HCT116^(hSMO)cells in vitro,compared with the control(P<0.05).Meanwhile,ursolic acid also induced apoptosis of HCT116^(hSMO)cells in vitro(P<0.05).Furthermore,SC79(Akt activator)enhanced the expressions of p-Akt,GLI1 and c-Myc,which could be abolis

关键词： ursolic acid colorectal cancer noncanonical Hedgehog signaling apoptosis