可疑翼手参皂苷ColochirosideA体外抑制多种瘤细胞株增殖的研究

目的：研究可疑翼手参体内三萜皂苷ColochirosideA（cA）的体外抗肿瘤活性。方法：用稻瘟霉分生孢子模型对CA进行生物活性初筛,分别用MTT、SRB法检测CA对5种体外培养肿瘤细胞（BEL-7402、HO-8910、MDA—MB-435、MDA—MB-468、A431）和2种体外培养人血管内皮细胞（HMEC、HUVEC）的增殖抑制活性。结果：cA对稻瘟霉显示较强的抑制活性,最小活性浓度（MMDC）〈7．9ug／ml;对5种体外培养人肿瘤细胞显示显著抑制增殖活性,IC50为1．75ug／ml-3．66ug／ml;对2种体外培养人血管内皮细胞均显示显著抑制增殖活性,IC50值分别为2．55ug／ml和5．63ug／ml。结论：CA体外对多种肿瘤细胞具有抑制活性,有进一步研究开发的前景。     

四种抗肿瘤药物对鼻咽癌HNE-1细胞株增殖抑制作用的研究

[目的]研究4种常用抗肿瘤药物（长春新碱、平阳霉素、顺铂、氟尿嘧啶）对鼻咽癌细胞的增殖抑制作用。[方法]采用MTT法检测不同浓度长春新碱、平阳霉素、顺铂、5-Fu对鼻咽癌HNE-1细胞株增殖抑制率。[结果]长春新碱对细胞抑制率〈20％的浓度为〈50μg／ml,平阳霉素对细胞抑制率〈20％的浓度〈40μg／ml,顺铂、5-Fu对细胞抑制率与浓度呈正相关。[结论]4种抗肿瘤药物对鼻咽癌HNE-1细胞株均存在不同程度的增殖抑制作用。     

人参皂苷CK对人肺腺癌A549细胞株的增殖抑制作用及其机制

目的:探讨人参皂苷CK对人肺腺癌A549细胞增殖的影响及其作用机制。方法:不同浓度的人参皂苷CK作用于人肺腺癌A549细胞株,MTT比色法测定细胞增殖抑制率;倒置显微镜观察细胞形态学变化;流式细胞术分析细胞周期及细胞凋亡;ELISA法检测细胞内源性VEGF含量的变化。结果:人参皂苷CK对A549细胞增殖有抑制作用,其抑制作用呈浓度和时间依赖关系;倒置显微镜下可观察到细胞明显的形态学改变;流式细胞仪可检测到实验组细胞出现明显的凋亡峰,且细胞周期被阻滞在G0/G1期;实验组细胞培养液中的VEGF低于对照组(P＜0.05),且随着人参皂苷CK浓度增加和作用时间的延长,VEGF的含量降低(P＜0.05)。结论:人参皂苷CK可抑制人肺腺癌A549细胞的增殖并诱导其凋亡,并能抑制其内源性分泌VEGF。     

人肺癌细胞株A549自分泌抑制因子的初步研究

本文应用人肺癌细胞株A549,在无血清的萃取条件培养液中,37℃,5％CO_2,培养8天以后,收集培养上清,经12000rpm30分钟,去除沉淀,上清装入透析袋,用聚乙二醇包埋,快速浓缩而成粗提物,即称为该细胞的自分泌抑制因子(ACIF),并对它的作用进行初步探讨。我们利用人源5种传代肿瘤细胞株A549、K562、HL60、Smmc7721、BT325为靶细胞,以不同浓度的ACIF进行生长抑制试验,采用台酚兰染色计数法及~3H-TdR掺入法,两法结果基本一致,对亲源细胞A549,则随浓度升高抑制作用增强,而对其它ACIF的生物学特性及其体内作用,有待研究。     

环氧化酶-2抑制剂塞来昔布联合顺铂对A549人肺腺癌细胞株增殖影响的体外实验研究

目的观察环氧化酶-2（COX-2）抑制剂塞来昔布（Celecoxib）与化疗药物顺铂（DDP）联合应用对A549人肺腙癌细胞增殖的影响。方法应用MTS法检测Celecoxib与DDP联用对人肺腺癌A549细胞增殖的影响。结果在一定浓度范围内，Celecoxib和DDP均可抑制A549人肺腺癌细胞的生长，其抑制作用呈量-效关系。Celecoxib（0．35mg／L）与DDP联用可增强对A549细胞生长的抑制作用，DDP浓度≥1mg／L时两者具有协同或相加作用；且在一定浓度范围内，Celecoxib与DDP联用对细胞的生长抑制作用呈时-效关系。结论Celecoxib与DDP联合可增强对A549人肺腺癌细胞的生长抑制效应。     

The marine triterpene glycoside frondoside A exhibits activity in vitro and in vivo in prostate cancer

Despite recent advances in the treatment of metastatic castration‐resistant prostate cancer (CRPC), outcome of patients remains poor due to the development of drug resistance. Thus, new drugs are urgently needed. We investigated efficacy, toxicity and mechanism of action of marine triterpene glycoside frondoside A (FrA) using CRPC cell lines in vitro and in vivo. FrA revealed high efficacy in human prostate cancer cells, while non‐malignant cells were less sensitive. Remarkably, proliferation and colony formation of cells resistant to enzalutamide and abiraterone (due to the androgen receptor splice variant AR‐V7) were also significantly inhibited by FrA. The marine compound caused cell type specific cell cycle arrest and induction of caspase‐dependent or ‐independent apoptosis. Up‐regulation or induction of several pro‐apoptotic proteins (Bax, Bad, PTEN), cleavage of PARP and caspase‐3 and down‐regulation of anti‐apoptotic proteins (survivin and Bcl‐2) were detected in treated cells. Global proteome analysis revealed regulation of proteins involved in formation of metastases, tumor cell invasion, and apoptosis, like keratin 81, CrkII, IL‐1β and cathepsin B. Inhibition of pro‐survival autophagy was observed following FrA exposure. In vivo, FrA inhibited tumor growth of PC‐3 and DU145 cells with a notable reduction of lung metastasis, as well as circulating tumor cells in the peripheral blood. Increased lymphocyte counts of treated animals might indicate an immune modulating effect of FrA. In conclusion, our results suggest that FrA is a promising new drug for the treatment of mCRPC. Induction of apoptosis, inhibition of pro‐survival autophagy, and immune modulatory effects are suspected modes of actions.     

PE, a new sulfated saponin from sea cucumber, exhibits anti-angiogenic and anti-tumor activities in vitro and in vivo

Here, we examined the in vitro and in vivo anti-angiogenesis and anti-tumor activities of PE, a new marine-derived compound. Inhibition of angiogenesis was assessed in vitro using proliferation, migration, adhesion, tube-formation and apoptosis assays in PE-treated HMECs and HUVECs. In vivo, CAM assays were used to assess inhibition effect of PE on physiological angiogenesis, and immunofluorescent microscopy was used to examine tumor microvessel density and apoptosis in PE-treated mouse tumor models. Finally, Western blotting analyses were performed to examine the effect of PE on VEGF signaling in HMECs. The results showed that PE inhibited proliferation of HMECs and HUVECs with IC50 values of 2.22 +/- 0.31 microM and 1.98 +/- 0.32 microM, induced endothelial cell apoptosis at concentrations <2 microM, induced dose-dependent suppression of cell migration, cell adhesion and tube formation in HMECs and HUVECs, and showed anti-proliferative activities against several tumor cell lines (IC50 values of approximately 4 microM). In vivo, PE (5 nM/egg) suppressed spontaneous angiogenesis in our CAM assay, and induced marked growth inhibition in mouse sarcoma 180 and hepatoma 22 models. Specifically, PE treatment reduced mouse sarcoma 180 tumor volume by triggering apoptosis of both tumor and tumor-associated endothelial cells, preferentially targeting on endothelial cells comparable with tumor cells. Finally, PE treatment suppressed the active (phosphorylated) forms of VEGFR2, Akt, ERK, FAK and paxillin, which are involved in endothelial cell survival, proliferation, adhesion and migration. Our results indicate that PE exerts an anti-angiogenic activity associated with inhibition of VEGFR2 signaling, and an anti-tumor activity associated with decreased proliferation of tumor cells and increased apoptosis of both endothelial cells and tumor cells.     

Antitumor activities of an anthraquinone fraction isolated from in vitro cultures of Ophiorrhiza rugosa var decumbens

The biological effects of the anthraquinone fraction (AQf) isolated from in vitro cultures of Ophiorrhiza rugosa Wall. var decumbens (Rubiaceae) were evaluated. AQf showed differential activity on reactive oxygen species; it mediated the generation of superoxide radical and inhibited hydroxyl radical and lipid peroxidation. No considerable nitric oxide scavenging activity was observed for AQf. The AQf induced 50% cytotoxicity in Ehrlich ascites carcinoma and Dalton's lymphoma ascites at concentrations of 130 and 60 μg/mL, respectively. It effectively reduced the inflammation induced by carrageenan in mice. An AQf concentration of 200 mg/kg body weight reduced solid tumor progression in mice. It also prolonged the life span of ascites tumor-bearing mice compared with control mice.     

多抗甲素对树突状细胞瘤苗体外抗肿瘤活性的影响

 目的 研究多抗甲素（Polyactin A,PA）对树突状细胞（Dendritic cell,DC ）瘤苗体外抗肿瘤活性的影响。 方法 用GM-CSF、IL-4、TNF-α体外诱导扩增脐血单个核细胞（CBMCs）中的DC,24h后将凋亡、坏死的HeLa、HepG2细胞分别加入CBMCs共孵育,使DC接受肿瘤细胞全抗原刺激,成为DC瘤苗,之后再加入PA刺激活化DC瘤苗,3天后以CBMCs为效应细胞（另设未加PA组为对照）,以相应的肿瘤细胞为靶细胞,用MTT法测定不同效靶比下效应细胞对HeLa、HepG2细胞的杀伤率。 结果加入PA后,CBMCs对HeLa、HepG2细胞的杀伤率显著提高,明显高于未加PA组。 结论PA可增强DC瘤苗对相应肿瘤的杀伤率。     

Antitumor activity of total paeony glycoside against human chronic myelocytic leukemia K562 cell lines in vitro and in vivo

To explore the molecular mechanisms of human leukemia cells by total paeony glycoside (TPG), which is extracted from the root of Radix Paeoniae Rubra. The viability of K562 cells was assessed by MTT assay. Flow cytometry was used to detect apoptosis and cell cycle analysis. The changes in intracellular Ca²⁺ concentration were determined by fluorescent dye Fluo-3, and mitochondrial membrane potential was determined by the retention of the dye Rh123. The cytoplasmic Bax, Bcl-xL, and Bcl-2 protein expressions were determined by western blot. The mRNA expression of caspase-3, caspase-8, and caspase-9 was detected by RT-PCR. K562 cells were subcutaneously inoculated into nude mice to study the in vivo antitumor effects of TPG. The growth of K562 cells was inhibited and arrested in G0/G1 phase by TPG. TPG also caused apoptosis in K562 cells evidenced by cytosolic accumulation of cytochrome c, caspase-9, and caspase-3. TPG could down-regulate Bcl-2 and Bcl-xL and up-regulate Bax in K562 cells. TPG showed a significant decreased tumor volume and tumor weight in nude mice inoculated with K562 cells. TPG can be developed as a promising anti-chronic myeloid leukemia drug.     

Antitumor activity of histone deacetylase inhibitor trichostatin A in osteosarcoma cells

Background: Histone deacetylase (HDAC) inhibitors have been reported to induce cell growth arrest,apoptosis and differentiation of tumor cells. The present study aimed to examine the effects of trichostatin A(TSA), one such inhibitor, on the cell cycle, apoptosis and invasiveness of osteosarcoma cells. Methods: MG-63 cells were treated with TSA at various concentrations. Then, cell growth and apoptosis were determinedby 3-(4, 5-dimethyl-2-thiazolyl)-2H-tetrazolium bromide (MTT) and TUNEL assays, respectively; cell cyclingwas assessed by flow cytometry; invasion assays were performed with the transwell Boyden Chamber system.Results: MTT assays revealed that TSA significantly inhibited the growth of MG-63 cells in a concentrationand time dependent manner. TSA treated cells demonstrated morphological changes indicative of apoptosisand TUNEL assays revealed increased apoptosis of MG-63 cells after TSA treatment. Flow cytometry showedthat TSA arrested the cell cycle in G1/G2 phase and annexin V positive apoptotic cells increased markedly. Inaddition, the invasiveness of MG-63 cells was inhibited by TSA in a concentration dependent manner. Conclusion:Our findings demonstrate that TSA inhibits the proliferation, induces apoptosis and inhibits invasiveness ofosteosarcoma cells in vitro. HDAC inhibitors may thus have promise to become new therapeutic agents againstosteosarcoma.     

Anticancer activities of sesquiterpene lactones from Cyathocline purpurea in vitro

Purpose

Cyathocline purpurea has been traditionally used to treat various diseases including cancers for many years. However, these applications of C. purpurea have not been supported by pharmacological investigation. The objective of this study is to investigate the anticancer activities of three main constituents such as santamarine, 9β-acetoxycostunolide and 9β-acetoxyparthenolide isolated from C. purpurea in vitro.

Methods

Cell viability was determined by trypan blue exclusion and methylene blue assays. Colony formation was assessed by microtitration cloning assay. DNA synthesis was determined by tritiated thymidine incorporation assay. Cell cycle analysis was carried out by flow cytometry. Apoptosis was observed by DAPI staining assay and Caspase 3/7 activities was measured using Caspase-Glo^® 3/7 assay kit.

Results

Santamarine, 9β-acetoxycostunolide and 9β-acetoxyparthenolide inhibited the growth of L1210 murine leukaemia, CCRF-CEM human leukaemia, KB human nasopharyngeal carcinoma, LS174T human colon adenocarcinoma and MCF 7 human breast adenocarcinoma cells in vitro, with IC₅₀ in the range of 0.16–1.3 μg/mL. In L1210 model, santamarine and 9β-acetoxycostunolide inhibited L1210 cell growth, colony formation and [³H]-thymidine incorporation in time- and concentration-dependent manners. Flow cytometry studies showed that santamarine and 9β-acetoxycostunolide blocked L1210 cells in the G₂/M phase of the cell cycle. DAPI staining and caspase activity assays showed santamarine and 9β-acetoxycostunolide induced apoptosis and activated caspase 3 in L1210 cells.

Conclusions

These results indicated that santamarine, 9β-acetoxycostunolide and 9β-acetoxyparthenolide exhibit significant anticancer activities in vitro. The inhibitory effects of santamarine and 9β-acetoxycostunolide on L1210 cells are cytotoxic rather than just cytostatic. They block mitosis and reduce uptake of thymidine. The mechanism of the cytotoxicity of santamarine and 9β-acetoxycostunolide to L1210 cells could be related to alkylation of the sulfhydryl enzymes involved in nucleic acids and protein synthesis, as previously found for other sesquiterpenes with the α-methylene-γ-lactone moiety present in santamarine, 9β-acetoxycostunolide and 9β-acetoxyparthenolide. It may also be related to suppression of microtubular proteins. Santamarine and 9β-acetoxycostunolide induced apoptosis of L1210 cells via activation of caspase 3.     

Antitumor activity of a polysaccharide from longan seed on lung cancer cell line A549 in vitro and in vivo

In this study, a water-soluble longan seed polysaccharide (WLSP), with a molecular weight of 57 kDa, was isolated from longan seed. Gas chromatography (GC) analysis showed that WLSP was composed mainly of rhamnose (Rha), mannose (Man), arabinose (Ara), galactose (Gal), and glucose (Glc), with molar ratios of 2.4:1.5:2.3:5.6:6.5. The result in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that WLSP showed a dose-dependent antiproliferative effect on the proliferation of A549 human lung cancer cells, which is consistent with the amount of lactate dehydrogenase (LDH) release from A549 cells. Prompted by this antiproliferative effect, we further examined its antiproliferative mechanism and in vivo anticancer effect. Our results showed that WLSP had the ability to cause cell cycle arrest in G1 phase, activation of caspases 3 and 9, and cleavage of poly[ADP (ribose)] polymerase (PARP) in A549 cells. The result of this in vivo study showed that WLSP could suppress the growth of xenograft A549 tumors and induce apoptosis. Taken together, these results indicate that WLSP exert an anticancer effect in vitro and in vivo and may be useful for the prevention of lung tumorigenesis.