Sesamin synergistically potentiates the anticancer effects of γ-tocotrienol in mammary cancer cell lines

γ-Tocotrienol and sesamin are phytochemicals that display potent anticancer activity. Since sesamin inhibits the metabolic degradation of tocotrienols, studies were conducted to determine if combined treatment with sesamin potentiates the antiproliferative effects of γ-tocotrienol on neoplastic mouse (+ SA) and human (MCF-7 and MDA-MB-231) mammary cancer cells. Results showed that treatment with γ-tocotrienol or sesamin alone induced a significant dose-responsive growth inhibition, whereas combination treatment with these agents synergistically inhibited the growth of + SA, MCF-7 and MDA-MB-231 mammary cancer cells, while similar treatment doses were found to have little or no effect on normal (mouse CL-S1 and human MCF-10A) mammary epithelial cell growth or viability. However, sesamin synergistic enhancement of γ-tocotrienol-induced anticancer effects was not found to be mediated from a reduction in γ-tocotrienol metabolism. Rather, combined treatment with subeffective doses of γ-tocotrienol and sesamin was found to induce G1 cell cycle arrest, and a corresponding decrease in cyclin D1, CDK2, CDK4, CDK6, phospho-Rb, and E2F1 levels, and increase in p27 and p16 levels. Additional studies showed that the antiproliferative effect of combination treatment did not initiate apoptosis or result in a decrease in mammary cancer cell viability. Taken together, these findings indicate that the synergistic antiproliferative action of combined γ-tocotrienol and sesamin treatment in mouse and human mammary cancer cells is cytostatic, not cytotoxic, and results from G1 cell cycle arrest.     

Immunomodulatory effects of proanthocyanidin A-1 derived in vitro from Rhododendron spiciferum

The objective of this study was to evaluate the immunomodulatory effects of proanthocyanidin A-1 (PAA-1) from leaves of Rhododendron spiciferum (Ericaceae). In vitro tests showed that PAA-1 stimulated cell proliferation of splenocytes and peritoneal macrophages significantly enhanced the cytotoxicity of natural killer (NK) cells and increased CD4⁺ and CD8⁺ cell populations. PAA-1 also regulated the expression of Th1- and Th2-related cytokines. Moreover, this study showed that PAA-1 exhibited a significant effect on NBT dye reduction and lysosomal enzyme activity responses in macrophages, indicating effective phagocytic activation. These results revealed that PAA-1 exhibits immunomodulatory activity without a clear dose response.     

Ganoderic acid DM, a natural triterpenoid, induces DNA damage, G1 cell cycle arrest and apoptosis in human breast cancer cells

Ganoderic acid DM (GADM) is a triterpenoid isolated from Ganoderma lucidum, a well-known edible medicinal mushroom. In the present study, we found that GADM effectively inhibited cell proliferation and colony formation in MCF-7 human breast cancer cells, which was much stronger than that of MDA-MB-231 breast cancer cells. GADM both concentration- and time-dependently mediated G1 cell cycle arrest and significantly decreased the protein level of CDK2, CDK6, cycle D1, p-Rb and c-Myc in MCF-7 cells. Moreover, GADM obviously induced DNA fragmentation and cleavage of PARP which are the characteristics of apoptosis and decreased the mitochondrial membrane potential in MCF-7 cells. Besides, we also showed that GADM elicited DNA damage as measured by comet assay which is a sensitive method for DNA damage detection. γ-H2AX, a marker of DNA damage, was also slightly up-regulated after treated with GADM for 6h, suggesting that the G1 cell cycle arrest and apoptosis induced by GADM may be partially resulted from GADM-induced DNA damage. These results have advanced our current understandings of the anti-cancer mechanisms of GADM.     

The effect of Astragaloside IV on immune function of regulatory T cell mediated by high mobility group box 1 protein in vitro

High mobility group box 1 protein (HMGB1), a potent pro-inflammatory cytokine, contributes to the pathogenesis of diverse inflammatory and infectious disorders. Some studies have illustrated the potential effect of HMGB1 on regulatory T cells (Tregs). Astragaloside IV (AST IV) isolated from a Chinese herb, Astragalus mongholicus, is known to have a variety of immunomodulatory activities. However, it is not yet clear whether AST IV possesses potential regulatory effect on the pro-inflammatory ability of HMGB1 with subsequent activation of Tregs. This study was carried out to investigate the antagonistic effects of different doses of AST IV on the immune function of Tregs mediated by HMGB1 in vitro. Tregs isolated from the spleens of mice were co-cultured with HMGB1 and/or AST IV. Cell phenotypes of Tregs were analyzed, and the contents of various cytokines in the cell supernatants as a result of co-culture and the proliferation of CD4⁺CD25⁻ T cells were determined. Results showed that HMGB1 stimulation resulted in significantly down-regulation of expressions of Tregs cell phenotypes. However, AST IV can rival the suppressing effect of HMGB1 on immune function of Tregs with a dose-dependent in vitro. These results indicate that AST IV has the potential therapeutic action on inflammation augmented by HMGB1.     

Antiproliferative activity of Saponaria vaccaria constituents and related compounds

Total methanolic extracts of Saponaria vaccaria seed derived from several varieties, as well as various purified components obtained through successive chromatographic separations of total extracts were evaluated for their growth inhibitory activity in WiDr (colon), MDA-MB-231 (breast), NCI-417 (lung) and PC-3 (prostate) human cancer cells as well as the non-tumorigenic fibroblast BJ (CRL-2522) cell line using MTT colorimetric assay. Purified bisdesmosidic saponins segetoside H and I were further examined using microscopy and apoptosis assays. Bisdesmosidic saponins exhibited dose-dependent growth inhibitory and selective apoptosis-inducing activity. Growth inhibitory effects were particularly strong in a breast (MDA-MB-231) and a prostate (PC-3) cancer cell line. Total extracts exhibited a different preference being most active against a colon cancer cell line (WiDr). In a comparison of varieties, all of the total seed extracts exhibited similar dose-dependent activities, but with some variation in potency. Monodesmosidic saponins vaccarosides A and B, phenolic vaccarin, and cyclopeptide segetalin A, co-occurring seed substituents, did not exhibit activity. The non-tumorigenic fibroblast cell line BJ (CRL 2522) was growth inhibited but did not undergo apoptosis when treated with bisdesmosidic saponins at low micromolar concentrations. Saponin-rich extracts from Kochia scoparia seed and Chenopodium quinoa were also evaluated alongside Saponaria saponins but did not exhibit activity. Closely related Quillaja saponins exhibited activity but were less potent.     

Estrogenic and anti-estrogenic activities of hispolon from Phellinus lonicerinus (Bond.) Bond. et sing

Hispolon was the main antitumor active ingredient in Phellinus sensu lato species. In order to confirm the dual regulating estrogenic ingredient and obtain more effective natural estrogen replacement drugs, hispolon was separated from Phellinus lonicerinus (Bond.) Bond. et sing. Hispolon exhibited significant anti-proliferative effect against estrogen-sensitive ER (+) MCF-7 cells in the absence of estrogen, and exhibits antagonistic effects on 17β-estradiol (E₂)-induced MCF-7 cell proliferation when E₂ and the different concentrations of hispolon were treated simultaneously. Hispolon also inhibited the proliferation of estrogen-negative ER (−) MDA-MB-231 cells at the concentration of 5.00 × 10^− 5 M. The yeast two-hybrid experiments showed that hispolon had strong and non-selective effects on the estrogen receptor (ER) α and ERβ at a concentration of 1.00 × 10^− 6 M. The ERβ-binding ability of hispolon was larger than ERα in the concentration range of 1.00 × 10^− 9 M and 1.00 × 10^− 7 M. Hispolon could increase the body weight coefficient, serum E₂ and progesterone contents in immature female mice at dose of 9.10 × 10^− 6 mol/kg, and increase coefficient of thymus and spleen in mice. The Gscores of hispolon-ERα and hispolon-ERβ docked complexes were − 7.93 kcal/mol and − 7.79 kcal/mol in docking simulations. Hispolon presented dual regulating estrogenic activities, which showed estrogenic agonist activity at low concentration or lack of endogenous estrogen, and the estrogenic antagonistic effect was stimulated at high concentrations or too much endogenous estrogen. Hispolon could be used for treating the estrogen deficiency-related disease with the benefit of non-toxic to normal cells, good antitumor effects and estrogenic activity.     

Echinacoside promotes bone regeneration by increasing OPG/RANKL ratio in MC3T3-E1 cells

Echinacoside (ECH), isolated from Cistanche tubulosa (Schrenk) R. Wight stems, was subjected to in vitro experiments to investigate its bioactivities on proliferation, differentiation and mineralization of the osteoblastic cell line MC3T3-E1. MTT assay, the alkaline phosphatase (ALP) activity and calcium deposition were determined, and the secretion of collagen I (COL I), osteocalcin (OCN), osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were also assayed by enzyme-linked immunosorbent assay (ELISA). The results showed that ECH caused a significant increase in cell proliferation, ALP activity, COL I contents, OCN levels and an enhancement of mineralization in osteoblasts at the concentration range from 0.01 to 10 nmol·L^− 1 (p < 0.05), suggesting that ECH has a stimulatory effect on osteoblastic bone formation or has potential activity against osteoporosis. In addition, the ratio of OPG/RANKL also could be enhanced by ECH. These findings provide the potent evidence that ECH can promote bone regeneration in cultured osteoblastic MC3T3-E1 cells, which might be done by elevating the OPG/RANKL ratio, and potential evidence for echinacoside to be a promising drug or a lead compound in the development of disease-modifying drug to prevent osteoporosis.     

Chemical constituents of Morus alba L. and their inhibitory effect on 3T3-L1 preadipocyte proliferation and differentiation

Mulberry leaf, an important traditional Chinese medicine, possesses many biological activities, including effects of anti-obesity. However, which constituents of mulberry leaf are responsible for its anti-adipogenic action is unclear. This study primarily investigated the chemical constituents from mulberry leaf and their bioactivity on the proliferation and differentiation of 3T3-L1 preadipocytes. A new flavane derivative, (2S)-4′-hydroxy-7-methoxy-8-prenylflavan (1), together with twelve known compounds including three flavanes (2–4), three chalcones (5–7), two flavones (8–9), two benzofurans (10–11) and two coumarin (12–13) was isolated from mulberry leaf. The structure of the new compound was elucidated by various spectroscopic methods including UV, HR-ESI-MS, ¹H and ¹³C NMR and CD. The results of activity screening showed that compound 2, 6 and 7 inhibited the proliferation and differentiation of 3T3-L1 preadipocytes.     

落叶松胚性愈伤组织诱导培养基中激素的311-A最优回归设计筛选

采用311-A最优回归设计，对落叶松胚性细胞诱导中激素种类与浓度优化筛选，建立胚性愈伤组织量依2，4-D、BA、KT的多基本原则 式回归方程，分析了试验因子的主效应和互作效应，借助此方程获得了3因子的最佳配比组合以及最佳胚性愈伤组织发和亘，2，4-D为1.29mg.L^-1、BA为0.39mg.L^-1和KT为0.58mg.L^-1时，落叶松胚性愈伤组织的诱导量可达到13.9317mg.个^-1(外植体）。结果表明，这是一种简便、实用、科学的优化培养基的途径。     

野生唐古特白刺悬浮细胞系的建立及生长特性

以野生唐古特白刺的茎段、叶片为外植体,研究了NaClO对外植体的消毒效果和激素2,4-D、NAA和6-BA对愈伤组织诱导及愈伤组织形态的影响,并利用疏松型愈伤组织建立了白刺悬浮细胞系,对其生长特征进行了分析。结果表明:对茎段和叶片用2%的NaClO消毒10 min最适宜,时间过长或过短均会影响成活率;3种激素对愈伤组织的相对生长量和形态均有影响,其中,2,4-D是主要影响因子,且茎段诱导出的愈伤组织相对生长量比叶片的高;根据愈伤组织的生长速度和形态,适宜用浅黄色疏松型愈伤组织构建唐古特白刺悬浮细胞系,最佳试验组合为MS+2,4-D 1.5 mg·L^-1+NAA 0.2 mg·L^-1+6-BA 0.4 mg·L^-1,最佳继代接种量为7.5 mL母液,生长曲线呈"S"型,培养第7天细胞分裂指数达最大值(5.1%),培养第3天细胞活力最强(吸光值为0.69),存活率在细胞生长延迟期和稳定期较对数期下降略快,但均保持在84% 93%间。     

A new cytotoxic flavonoid from the fruit of Sinopodophyllum hexandrum

Constituents of the fruit of Sinopodophyllum hexandrum (Royle) Ying (Sinopodophylli Fructus) were investigated. A new flavonoid, 8,2′-diprenylquercetin 3-methyl ether along with 9 known compounds were isolated and identified. Among them, the new compound 8,2′-diprenylquercetin 3-methyl ether exhibited cytotoxic activities against MDA-231 and T47D breast cancer cell lines, quercetin, kaempferol and rutin were isolated from Sinopodophylli Fructus for the first time.     

Hormone-sensitive lipase is involved in the action of hydroxysafflor yellow A (HYSA) inhibiting adipogenesis of 3T3-L1cells

BackgroundSafflor yellow A (SY) has been demonstrated to be beneficial to cardiovascular system. Our previous study showed that hydroxysafflor yellow A (HSYA), a main component of SY, could increase peroxisome proliferator-activated receptor γ mRNA expression. In this study, we investigate the effect of HSYA on the proliferation and adipogenesis of mouse 3T3-L1 preadipocytes.MethodsThe proliferation and adipogenesis of 3T3-L1 cells treated with HSYA was studied by 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) spectrophotometry, Oil Red O staining and intracellular triglyceride assay methods. HSL mRNA expression and promoter activity were studied by real-time quantitative RT-PCR, transient transfection and dual luciferase reporter gene methods.ResultsHSYA (0.1 mg/L) significantly inhibited the proliferation of 3T3-L1 cells when compared with control cells in 8 h. This effect was further enhanced with the extension time (24 to 96 h) and an increase of concentration of HSYA (1–10 mg/L). The maximal inhibitory action was observed at 0.1 mg/L HSYA in 72 h (86 ± 11.8% vs. 100 ± 4.1%, p < 0.01). HSYA notably reduced the amount of intracellular lipid and triglyceride content in adipocytes to 85% (1 mg/L) and 75% (100 mg/L) on Day 4 following the differentiation, respectively, while increased HSL mRNA expression and promoter activities to 2.7 fold and 1.55 fold, respectively (p < 0.01), in differentiated 3T3-L1 adipocytes.ConclusionsHSYA inhibits the proliferation and adipogenesis of 3T3-L1 preadipocytes. The inhibitory action of HYSA on adipogenesis may be due to the promotion of lipolytic-specific enzyme HSL expression by increasing HSL promoter activity.     

Triterpenes and steroids from the leaves of Aglaia exima (Meliaceae)

A study on the leaves of Aglaia exima led to the isolation of one new and seven known compounds: six triterpenoids and two steroids. Their structures were elucidated and analyzed mainly by using spectroscopic methods; 1D and 2D NMR, mass spectrometry, UV spectrometry and X-ray. All the triterpenoids and steroids were measured in vitro for their cytotoxic activities against eight cancer cell lines; lung (A549), prostate (DU-145), skin (SK-MEL-5), pancreatic (BxPC-3), liver (Hep G2), colon (HT-29), breast (MCF-7) and (MDA-MB-231). The new cycloartane triterpenoid, 24(E)-cycloart-24-ene-26-ol-3-one 1, showed potent cytotoxic activity against colon (HT-29) cancer cell line (IC₅₀ 11.5 μM).     

Effect of alcoholic extract of Ayurvedic herb Tinospora cordifolia on the proliferation and myeloid differentiation of bone marrow precursor cells in a tumor-bearing host

The present study investigates the effect of in vivo administration of alcoholic extract of Tinospora cordifolia whole plant (ALTC) on the proliferation and myeloid differentiation of bone marrow hematopoietic precursor cells in mice bearing a transplantable T cell lymphoma of spontaneous origin designated as Dalton's lymphoma (DL). BMC obtained from ALTC administered DL-bearing mice showed an enhanced BMC proliferation and colony forming ability in vitro in response to L929 conditioned medium as a source of colony stimulating factor (CSF). The number of granulocyte-macrophages colony (CFU-GM) was predominantly higher in the cultures of BMC obtained from ALTC administered mice as compared to mice injected with PBS alone. An increase in the count of bone marrow derived macrophages (BMDM) from ALTC administered mice was also observed along with an increase in the count of tumor associated macrophages. The BMDM obtained from ALTC administered mice showed an enhanced response to signal of LPS for activation to produce IL-1 and TNF. This study indicates that the T. cordifolia can influence the myeloid differentiation of bone marrow progenitor cells and the recruitment of macrophages in response to tumor growth in situ.     

Cytotoxic dimeric indole alkaloids from Catharanthus roseus

Three new dimeric indole alkaloids (1-3), together with five known ones (4-8), were isolated from the whole plants of Catharanthus roseus. The structures and absolute configurations of new compounds were elucidated by means of NMR and CD analyses. All these compounds were evaluated for their in vitro cytotoxic activities against human breast cancer cell line MDA-MB-231.     

Effect of <Emphasis Type="Italic">Pinus radiate</Emphasis> bark extracts with different molecular weight distributions on cell growth of NIH/3T3 fibroblasts and dendrite retraction of B16 melanoma cells

Hot water extract (HWE) of Pinus radiata bark was separated into monomeric polyphenol (MPP), oligomeric proanthocyanidin (OPA), and polymeric proanthocyanidin (PPA) fractions by monitored chromatography using a Sephadex LH 20 column and an UV detector at 250 nm. The effects of these fractions on NIH/3T3 fibroblasts and B16 melanoma cells were examined by evaluating cell viability, melanogenesis (melanin content), morphological changes, and tyrosinase inhibitory activity. The polyphenolic fractions had a proliferation effect on fibroblasts, with cell growth increasing significantly (P < 0.01) even at the high concentration of 1250 μg/ml. At 125 μg/ml, HWE, MPP, and OPA had no effect on melanogenesis, whereas PPA significantly upregulated melanogenesis (P < 0.05). Melanogenesis was significantly upregulated in melanoma cells treated with these fractions at the high concentration of 600 μg/ml (P < 0.01). B16 melanoma cells cultured with the proanthocyanidin (PA)-rich fractions (HWE, OPA, and PPA) showed marked dendrite retraction, leading to significant morphological transformation. OPA in particular showed colored adhesion on the surface of melanoma cells. All four fractions significantly inhibited mushroom tyrosinase activity when compared to arbutin and ascorbic acid 2-glucoside (P < 0.05). The tyrosinase inhibitory activity of OPA and PPA did not differ significantly (P = 0.11), indicating that the inhibitory effects had a low correlation with molecular weight distribution. The inhibition kinetics of PPA determined using a Lineweaver-Burk plot indicated that PPA is a noncompetitive inhibitor of l-3,4-dihydroxyphenylalanine oxidation by mushroom tyrosinase.     

Somatic embryogenesis from cell suspension cultures of aspen clone

Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions.     

几种丛生竹愈伤组织诱导与防褐变技术研究

利用小佛肚竹、凤尾竹和孝顺竹幼竹的茎尖和带节茎段研究了愈伤组织诱导和控制褐变的方法.试验结果表明:在3种生长调节剂(2,4-D、KT、IBA)的组合中,以2,4-D(2～3 mg·L-1)效果最好,愈伤组织的诱导率最高,大部分外植体都能诱导出愈伤组织,生长良好;2,4-D 2 mg·L-1 KT0.5 mg·L-1能使部分茎段诱导出愈伤组织,而茎尖诱导的愈伤组织水渍化,易褐变;KT 4 mg·L-1能使个别茎段有愈伤组织产生,但易褐化;KT 4 mg·L-1 IBA0.5 mg·L-1能诱导出愈伤组织,但生长缓慢.添加抗氧化剂控制愈伤组织褐变的效果要优于吸附剂,其防褐化能力:抗坏血酸(5 mg·L-1)＞半胱氨酸(100 mg·L-1)＞PVP(1 g·L-1)＞活性炭(1 g·L-1).在培养基中加入抗坏血酸(5 mg·L-1),使小佛肚竹、凤尾竹、孝顺竹的茎尖和茎段的褐变率分别比对照下降了51.3%、43.3%、36.8%和62.7%、42.7%、30.8%.外植体在无菌水或半胱氨酸(100 mg·L-1)溶液中浸泡2～4 h也有利于控制褐化.暗培养有利于愈伤组织生长,对控制褐化也有一定作用.