A new cerebroside and its anti-proliferation effect on VSMCs from the radix of Cyperus rotundus L.

A new cerebroside,1-O-(β-D-glucopyranosyloxy)-(2S,3R,4E,8Z)-2-[(2′R)-2’-hydroxylignoceranoylamino]-4,8-tetradecene-3- diol was isolated from the 60%EtOH extract of traditional Chinese medical plant Cyperus rotundus L.Its structure was determined on the basis of spectroscopic data.This new compound showed anti-proliferation effect on vascular smooth muscle cells(VSMCs).

Effects of Ligustrazine Combined with Cisdichlorodiamine Platinum on Anti-proliferation,Cell Cycle and Apoptosis of Human Gastric Carcinoma SGC-7901 Cell Lines

[Objective] This study aimed to investigate the combination effects of ligustrazine and cis-dichlorodiamine platinum （DDP） on anti-proliferation, cell cycle and apoptosis of SGC-7901 cell lines in vitro. [Methed] SGC-7901 cells were treat- ed with ligustrazine and DDP alone or combined for 48 h for morphology assay. Anti-proliferative effects with the same treatment for 24, 48 and 72 h were assayed by MTT method, respectively. Cell cycle distribution and apoptosis assay of treated cells were performed in flow cytometry. Zhengjun Jin＇s protocol was used to assay the effect of drug combination. [Result] Ligustrazine significantly increased the prolif- eration inhibition rate of DDP on SGC-7901 cells in combination with DDP, com- paring with the effects of ligustrazine or DDP alone, and exhibited synergistic antitu- mor effect. The combination drug treatment induced cell cycle arrest occurred in S and G2 phase of the cell cycle and increased the apoptosis rate significantly. [Conclusion] Our results indicate that ligustrazine, as a low-toxic and natural herbal component, can significantly increase the anti-proliferative effect and apoptosis rate of antitumor drug DDP on human gastric carcinoma SGC：.-7901 cells.

Anti-nitric oxide production, anti-proliferation and antioxidant effects of the aqueous extract from Tithonia diversifolia

Objective: To determine the cytotoxicity, reduction in nitric oxide production and antioxidative activity of the aqueous leaf extract from Tithonia diversifolia(T. diversifolia) in an in vitro model.Methods: Leaves of T. diversifolia were collected from natural habitats and extracted with distilled water using the decoction method. The cytotoxic effect of the extract in terms of cell viability was determined using RAW264.7 cells and human peripheral blood mononuclear cells(PBMCs) via the mitochondrial respiration method using the MTT reagent. The effect of the extract on lipopolysaccharide(LPS)-induced nitric oxide production in RAW264.7 cells was measured using the Griess reagent. The chemical antioxidant was evaluated by ABTS- and DPPH-radical scavenging assays.Results: The half-maximal cytotoxic concentration values were 145.87 mg/m L and73.67 mg/m L for human PBMCs and RAW264.7 cells, respectively. In the presence of phytohemagglutinin-M, the IC_(50) on PBMCs proliferation was 4.42 mg/m L. The noncytotoxic range of the extracts inhibited LPS-induced nitrite production in RAW264.7 cells with an IC_(50) value of 11.63 mg/m L. To determine the anti-oxidative properties, the N-acetyl cysteine equivalent antioxidant capacity of the extract was(32.62 ± 1.87) and(20.99 ± 2.79)mg N-acetyl cysteine/g extract, respectively determined by the ABTS-radical and DPPHradical assay. However, the extract did not confer death protection in a hydrogen peroxideinduced RAW264.7 co-culturing model.Conclusions: Our study demonstrated the immunomodulation caused by the aqueous leaf extract of T. diversifolia, resulting from the inhibition of phytohemagglutinin-Minduced PBMCs proliferation and LPS-induced nitric oxide production in RAW264.7macrophages. Although the anti-oxidative activity was presented in the chemical-based anti-oxidant assay, the extract cannot protect cell death from stress conditions.

Anti-proliferation activity of terpenoids isolated from Euphorbia kansui in human cancer cells and their structure-activity relationship

Euphorbia kansui is a commonly used traditional Chinese medicine for the treatment of edema,pleural effusion,and asthma,etc.According to the previous researches,terpenoids in E.kansui possess various biological activities,e.g.,anti-virus,anti-allergy,antitumor effects.In this work,twenty five terpenoids were isolated from E.kansui,including thirteen ingenane-and eight jatrophane-type diterpenoids(with two new compounds,kansuinin P and Q)and four triterpenoids.Eighteen of them were analyzed by MTS assay for in vitro anticancer activity in five human cancer cell lines.Structure-activity relationship for 12 ingenane-type diterpenoids in colorectal cancer Colo205 cells were preliminary studied.Significant anti-proliferation activities were observed in human melanoma cells breast cancer MDA-MB-435 cells and Colo205 cells.More than half of the isolated ingenane-type diterpenoids showed inhibitory activities in MDA-MB-435 cells.Eight ingenane-and one jatrophane-type diterpenoids possessed much lower IC_(50) values in MDA-MB-435 cells than positive control staurosporine.Preliminary structure-activity relationship analysis showed that substituent on position 20 was important for the activity of ingenane-type diterpenoids in Colo205 cells and substituent on position 3 contributed more significant biological activity of the compounds than that on position 5 in both MDA-MB-435 and Colo205 cells.

Toxicarioside A inhibits SGC-7901 proliferation,migration and invasion via NF-κB/bFGF signaling

AIM:To investigate the inhibitory role of toxicarioside A on the gastric cancer cell line human gastric cancer cell line(SGC-7901) and determine the underlying molecular mechanism.METHODS:After SGC-7901 cells were treated with toxicarioside A at various concentrations(0.5,1.5,4.5,9.0 μg/mL) for 24 h or 48 h,cell viability was determined by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl2H-tetrazolium bromide assay,and the motility and invasion of tumor cells were assessed by the Transwell chamber assay.Immunofluorescence staining,reverse transcription polymerase chain reaction and Western blotting were performed to detect the expression of basic fibroblast growth factor(bFGF) and fibroblast growth factor receptor-1(FGFR1),and nuclear factorkappa B(NF-κB) activation was examined by electrophoretic mobility shift assay.RESULTS:The results showed that toxicarioside A was capable of reducing cell viability,inhibiting cell growth,and suppressing cell migration and invasion activities in a time-and dose-dependent manner in SGC-7901 cells.Further analysis revealed that not only the expression of bFGF and its high-affinity receptor FGFR1 but also the NF-κB-DNA binding activity were effectively blocked by toxicarioside A in a dose-dependent manner compared with the control group(P < 0.05 or P < 0.01).Interestingly,application of the NF-κB specific inhibitor,pyrrolidinedithiocarbamate(PDTC),to SGC-7901 cells significantly potentized the toxicarioside A-induced down-regulation of bFGF compared with the control group(P < 0.05).CONCLUSION:These findings suggest that toxicarioside A has an anti-gastric cancer activity and this effect may be achieved partly through down-regulation of NF-κB and bFGF/FGFR1 signaling.

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck,one of the most popular edible mushrooms,has attracted great attention due to its delicious taste and healthy properties.However,both its biological effects and the possible mechanism of action have not yet been known.We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck(MPE)against HepG2 human hepatocellular carcinoma cells.Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90%(P<0.01).The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax,Bcl-2 and cleaved caspase-3.On the other hand,MPE might trigger cell cycle arrest at G0/G1/S phases by managing p21,Cyclin D1,cyclin-dependent kinases-4(CDK4)and proliferating cell nuclear antigen(PCNA).Additionally,MPE downregulated TRAF-2 and p-p53,while upregulated p-ASK1 and p-p38.Therefore,it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.

Phylogenetic analysis and protective effects of thymol and its chromatographic fractions from a novel wild mushroom in combating oxidative stress

Mushrooms are good sources of phytochemicals that have antioxidant and anti-proliferative effects.This study identifi ed a unique isoform of 18S rRNA gene(864 bp)from a novel wild mushroom(SMK-1)(GenBank accession number:SUB3267363).Thin layer chromatographic(TLC)profiling of the methanolic extract of the dried mushroom fruiting bodies of SMK-1 revealed the presence of phenolic and fl avonoid fractions with retention factor(Rf)values of 0.955 and 0.927 respectively.The GC/MS chromatograms of the SMK-1 methanolic extract identifi ed the main bioactive compound was phenol,5-methyl-2-(1-methylethyl)(74.00%)(thymol).The radical scavenging activity for the fl avonoid fraction was greater than the phenolic fractions(Rf–phenolics fractions>Rf–fl avonoid fractions)with the antioxidant activity more than that of standard ascorbic acid.Also,the phenolic and flavonoid fractions of SMK-1 expressed cytotoxic effects in HeLa cells with IC50 values ranging from 5μg/mL to 80μg/mL in a dose-dependent manner.This present research highlights the presence of high thymol concentration in a novel wild mushroom that has antioxidant and anti-poliferative potential with therapeutic benefi ts.The application of thymol natural products from novel mushroom SMK-1 as nutrition supplements could inhibit oxidative stress triggered by numerous pathologies that may pave the way to develop a new therapeutic natural drug.

Essential oil from Saussurea costus inhibits proliferation and migration of Eca109 cells via mitochondrial apoptosis and STAT3 signaling

Objective:To investigate the effect and its underlying molecular mechanisms of essential oil from Saussurea costus in esophageal cancer cell line Eca109.Methods:The chemical composition of essential oil from Saussurea costus was investigated by gas chromatography-mass spectrometry(GC-MS).The anti-proliferative,anti-migrative,and apoptotic effects of essential oil from Saussurea costus against Eca109 cells were analyzed.Moreover,the expression of proteins associated with cell cycle,metastasis,and apoptosis was determined.Results:GC-MS analysis showed that essential oil from Saussurea costus was predominantly comprised of sesquiterpenes.Saussurea costus essential oil inhibited the viability of Eca109 cells in a dose-and time-dependent manner with IC_(50) values of(24.29±1.49),(19.16±2.27)and(6.97±0.86)μg/mL at 12,24,and 48 h,respectively.The expression levels of target proteins in the cell cycle(phase G_(1)/S),including cyclin D1,p21,and p53,were affected by Saussurea costus essential oil.The essential oil also downregulated the expression of metastasis-related proteins MMP-9 and MMP-2.Moreover,it induced apoptosis of Eca109 cells through the mitochondrial pathway,as well as inhibition of STAT3 phosphorylation.Conclusions:The essential oil from Saussurea costus exhibited anti-proliferative,anti-migrative,and apoptotic effects on Eca109 cells,and could be further explored as a potential anti-esophageal cancer agent.

Role of metabolomics assisting the pharmacological activity and molecular mechanism in Panax notoginseng

OBJECTIVE The strategy and techniques of metabolomics was applied for the pharmacology and molecular mechanism research of Panax notoginseng(PN)in traditional Chinese medicine.METHODS The global metabolic profiles of PN were investigated by the NMR-based metabolomics.The different parts of PN were scanned into metabolic profiles by 1H-NMR.The significant differences of these metabolic profiles were analyzed by PCA,PLS-DA,PLS-R,etc.The pharmacological effects including free radical scavenging activity(FRSA),anti-proliferation to human colorectal cancer cell line(HCT116),xanthine oxidase inhibition,were followed in vitro.Additionally,the molecular mechanism of xanthine degrading process by PN was attempted by 1H-NMR.RESULTS The NMR-based metabolic profiles of different parts(upper part of root,middle part of root,lower part of root,hairy root,leaf and stem)of PN presented significant differences by multivariate statistical analysis.The hairy root and leaf revealed highest anti-proliferative effect to HCT116;the leaf and stem of PN showed highest level of FRSA;the leaf,stem,hairy root effected the xanthine degrading 1 metabolic pathway.And the H-NMR based molecular mechanism experiment showed that the xanthine metabolic pathway degraded by PN depended on the direct inhibition to xanthine.CONCLUSION The metabolomics strategy provided complementary chemical profiling to medicinal herbs,which accelerated the development of pharmacology and mechanism of action in traditional medicine.The subsidiary parts of PN,as leaf,stem and hairy root,have the potential to develop new drugs in curing cancer,inflammation and gout.

Lunasin peptide promotes lysosome-mitochondrial mediated apoptosis and mitotic termination in MDA-MB-231 cells

Lunasin,a novel bioactive peptide,is well-known for its anti-proliferation activity.However,the mechanism of this effect is still poorly reported.Here,synthesized lunasin was used and its anti-proliferative function was observed at the concentration of 0.25 mg/m L in human breast cancer cell MDA-MB-231.Conjoint analysis of transcriptome and proteome of MDA-MB-231 cells was further performed.The results demonstrated that cysteinyl aspartate specific proteinase(CASP)3,CASP 7,and CASP 14 were significantly up-regulated after lunasin exposure,together with an increased Bax/Bcl-2 ratio from 22.9 to 210.6,which indicated that caspase-mediated mitochondria intrinsic apoptosis was highly activated.Moreover,lysosomal pathway was signifi cantly suppressed under lunasin exposure,suggesting that lysosome may cooperate with mitochondria to participate in apoptosis.In addition,lunasin also down-regulated genes involved in DNA replication in MDA-MB-231 cells.Overall,our study reveals that the anti-proliferation effect of lunasin peptide might be triggered via the inhibition of DNA replication and cell mitosis,as well as the promotion of lysosome-mitochondrial mediated cell apoptosis.

Extraction of Phenolic Compounds from Olive Leaf Extracts and Their Effect on Proliferation of Human Carcinoma Cell Lines

The aim of this work was to evaluate the effect of different olive leaf extracts (OLE) from different leaf growing stages on human carcinoma cell lines. OLE were tested in human carcinoma cell lines in vitro and cells were plated in 96-microtiter culture plates for each OLE concentration. Fresh (F) and freeze-dried (FD) leaves exhibited phenolic compounds in the range of 2.09 ± 0.10 to 8.44 ± 0.64 and 7.72 ± 0.56 to 24.65 ± 1.9 mg gallic acid equivalents/g leaves, respectively. OLE from several Portuguese olive tree cultivars were found to inhibit the growth of human carcinoma cell lines in a range of 2.09 - 8.44 μg phenolic compound/well (209 - 844 μg/ml) and 0.07 - 2.40 μg phenolic compounds/well (7 - 240 μg/ml) for fresh and freeze-dried leaves, respectively. Young (Y) leaves have revealed the highest cell growth inhibition ranging from about 95% for Cobran?osa, followed by 90% for Cobran?osa, 90% for Arbequina and 75% for Arbequina for cell lines A549, HeLa, A431 and OE21, respectively. The lowest cell growth inhibition (35%) was observed for Galega (Y) leaf extract on cell line A549. However, FD samples exhibited a distinctive pattern since cell growth inhibition was highest at highest extract dilution tested, for A431 (Galega Y) followed by A549 (Cobran?osa Y) with cell inhibition of 75% and 70%, respectively. The data presented in this work strongly suggest that OLEs inhibit the growth of human carcinoma cell lines.

Melatonin: A Powerful Integrative Adjunctive Agent for Oncology

Melatonin is an established hormone supplement and has been well recognized for its effect on the circadian cycle to improve sleep, REM (rapid eye movement), and aiding in jetlag recovery. The utility of melatonin extends beyond sleep aid, however. This hormone also possesses less well-known antioxidant action and even robust anticancer activity. Melatonin may be a key supplement for addressing age-related neurologic decline while serving as a valuable adjunctive cancer treatment that reduces drug resistance in tumors and downregulates angiogenesis. In immunotherapy, melatonin activates Natural Killer (NK) cells nested within tumoral tissue and does not have the side effect profile of other immunoreactive agents used for chemotherapy. Since melatonin is found in high concentrations in the brain and other hormone-linked tissues, the relevance of melatonin is increased for the treatment of estrogen-linked cancers. The immunomodulatory effect of melatonin may also help with chronic inflammation seen in patients with autoimmune disorders. All of these effects together represent a unique and versatile therapeutic agent for integrative medicine. No other commercially available drug possesses all of these therapeutic mechanisms while having a very minimal side effect profile and being considered overall to be safe to use. Currently, melatonin is underutilized in medicine, especially in the field of integrative oncology and represents a crucial supportive adjuvant to improve the lives of patients.

Butyrate-induced GPR41 Activation Inhibits Histone Acetylation and Cell Growth

Butyrate has been recently identified as a natural ligand of the G-protein-coupled receptor 41 （GPR41）. In addition, it is an inhibitor of histone deacetylase （HDAC）. Butyrate treatment results in the hyperacetylation of histones, with resultant multiple biological effects including inhibition of proliferation, induction of cell cycle arrest, and apoptosis, in a variety of cultured mammalian cells. However, it is not clear whether GPR41 is actively involved in the above-mentioned processes. In this study, we generated a stable cell line expressing the hGPR41 receptor in order to investigate the involvement of GPR41 on butyrate-induced biochemical and physiologic processes. We found that GPR41 activation may be a compensatory mechanism to counter the increase in histone H3 acetylation levels induced by butyrate treatment. Moreover, GPR41 had an inhibitory effect on the anti-proliferative, pro-apoptotic effects of butyrate. GPR41 expression induced cell cycle arrest at the Gl-stage, while its activation by butyrate can cause more cells to pass the G1 checkpoint. These results indicated that GPR41 was associated with histone acetylation and might be involved in the acetylation-related regulation of cell processes including proliferation, apoptosis, and the cell cycle.

Multi-omics approaches identify SF3B3 and SIRT3 as candidate autophagic regulators and druggable targets in invasive breast carcinoma

Autophagy is a critical cellular homeostatic mechanism,and its dysfunction is linked to invasive breast carcinoma(BRCA).Recently,several omics methods have been applied to explore autophagic regulators in BRCA;however,more reliable and robust approaches for identifying crucial regulators and druggable targets remain to be discovered.Thus,we report here the results of multi-omics approaches to identify potential autophagic regulators in BRCA,including gene expression(EXP),DNA methylation(MET)and copy number alterations(CNAs)from The Cancer Genome Atlas(TCGA).Newly identified candidate genes,such as SF3 B3,TRAPPC10,SIRT3,MTERFD1,and FBXO5,were confirmed to be involved in the positive or negative regulation of autophagy in BRCA.SF3 B3 was identified firstly as a negative autophagic regulator,and siRNA/shRNA-SF3 B3 were shown to induce autophagyassociated cell death in in vitro and in vivo breast cancer models.Moreover,a novel small-molecule activator of SIRT3,1-methylbenzylamino amiodarone,was discovered to induce autophagy in vitro and in vivo.Together,these results provide multi-omics approaches to identify some key candidate autophagic regulators,such as the negative regulator SF3 B3 and positive regulator SIRT3 in BRCA,and highlight SF3 B3 and SIRT3 as new druggable targets that could be used to fill the gap between autophagy and cancer drug development.

A new saikogenin from the roots of Bupleurum bicaule

AIM: To study the chemical constituents from the roots of Bupleurum bicaule Helm(Apiaceae). METHOD: Silica gel, Sephadex LH-20, MPLC Rp-C18 column chromatography, and HPLC were used for isolation of compounds. The structures were elucidated on the basis of 1D- and 2D-NMR technology and HRESI-MS. Compounds were evaluated in vitro for their inhibitory ability against the proliferation of rat mesangial cells by the MTT method. RESULTS: Twelve compounds were isolated, and their structures were identified on the basis of their spectroscopic and ico-chemical properties as 13, 28-epoxy-olean-11-en-3-one(1), saikogenin E(2), saikogenin G(3), 11α-methoxy-3β, 16β, 23, 28-tetrahydroxyolean-12-ene(4), saikogenin D(5), prosaikogenin F(6), prosaikogenin A(7), prosaikogenin G(8), prosaikogenin D(9), laccaic acid(10), methyl gallate(11), and ethyl gallate(12). Compounds 1, 2, 7, 8, and 10 were observed to have inhibitory activity against mesangial cell proliferationin to different degrees. CONCLUSION: Compound 1, 8, and 10 exhibit significant inhibitory effects on rat mesangial cell proliferation induced by Ang II.

GKK1032B from endophytic Penicillium citrinum induces the apoptosis of human osteosarcoma MG63 cells through caspase pathway activation

Chemical investigation of the culture extract of an endophytic Penicillium citrinum from Dendrobium officinale,afforded nine citrinin derivatives(1–9)and one peptide-polyketide hybrid GKK1032B(10).The structures of these compounds were determined by spectroscopic methods.The absolute configurations of 1 and 2 were determined for the first time by calculation of electronic circular dichroism(ECD)data.Among them,GKK1032B(10)showed significant cytotoxicity against human osteosarcoma cell line MG63 with an IC50 value of 3.49μmol·L–1,and a primary mechanistic study revealed that it induced the apoptosis of MG63 cells via caspase pathway activation.

New Synthetic Route of Two Active Isomeric Metabolites of Erlotinib and Their Bioactivity Studies against Several Tumor Cell Lines

The synthesis and differential antiproliferative activity of two active isomeric metabolites of erlotinib were in- vestigated. This synthetic process had demonstrated to avoid the unstable 4-chloroquinazoline intermediates and long procedures. New intermediates and final compounds were identified by IH NMR, 13C NMR and ESI-TOF MS, and their purities were determined by high performance liquid chromatography. In vitro proliferative assay indi- cated that these two metabolites possessed antiproliferative activity against some conventional tumor cell lines and EGFR tyrosine kinase over-expression tumor cell lines as compared to erlotinib control, and their antitumor activity in cellular level was first reported here.

A comparison of in vitro anticancerous activity and mechanism of ethanolic extracts from different Ganoderma genus

Five ethanolic extracts from the mycelia of Ganoderma lucidum,G.tsugae,G.oerstedii,G.subamboinense,and G.resinaceum were respectively studied on their anticancerous activities against leukemic HL-60 cell line in vitro.Results showed that all five extracts potently inhibited HL-60 proliferation.The extract from G.lucidum mycelia exerted the highest activity.Annexin V/PI bivariate flow cytometric analysis further revealed that the five extracts significantly induced early apoptosis in HL-60 cells.The results illustrate that not only G.lucidum but also other Ganoderma species can inhibit cancer cells,and their mechanisms are related to induction of apoptosis.