4-叔丁基-5-(12,4,-三唑-1-基)-2-苄亚氨基噻唑的体外抗癌活性研究

目的研究4-叔丁基-5-(1,2,4-三唑-1-基)-2-苄亚氨基噻唑类衍生物对宫颈癌细胞系Hela、肝癌细胞系Bel 7402、鼻咽癌细胞系CNE 2的体外抗癌活性。方法细胞抑制率采用四甲基偶氮唑蓝(MTT)法测定。每种试样设置5个浓度梯度(0.025、0.05、0.10、.25、0.5μmol/ml),每个浓度取4个平行样,并设置无药对照实验,用半数抑制浓度(IC50)评价目标化合物的抗癌活性。结果被测化合物对3种癌细胞均表现出一定的抑制作用,其中化合物5对Hela癌细胞和Bel 7402癌细胞的IC50值分别为0.076 1、0.062 8μmol/ml;化合物6对CNE 2癌细胞的IC50值为0.050 8μmol/ml。结论生物活性测试表明该类型氨基噻唑衍生物具有较好的抗癌活性,值得进一步研究和关注。     

氨甲喋呤对映体对ECV304细胞抑制作用及其机制

目的探讨氨甲喋呤[(±)MTX]及其对映体[(+)MTX、(-)MTX]对ECV304细胞的生长抑制作用并探讨其机制。方法采用培养的ECV304细胞,应用MTT比色法分析其活性;用光学显微镜观察细胞的形态学变化;碘化丙啶(PI)单染流式细胞术检测细胞周期。结果在0.1～150μmol·L-1范围内,(+)MTX、(-)MTX和(±)MTX作用于ECV304细胞24、48、72h,均抑制细胞ECV304增殖,但抑制强度依次为(+)MTX>(±)MTX>(-)MTX,倒置显微镜观察不同浓度(±)MTX、(+)MTX和(-)MTX作用ECV304细胞不同时间后,出现细胞体积变小、核固缩等形态学改变。用10μmol·L-1的(+)MTX、(-)MTX和(±)MTX作用ECV304细胞48h后,PI单染流式细胞术检测ECV304细胞周期的影响,表明MTX消旋体及单一体干扰ECV304细胞DNA合成。结论(+)MTX和(-)MTX对ECV304细胞的抗增殖作用具有化学结构的立体选择性,(+)MTX的抗ECV304细胞增殖作用明显强于(-)MTX。     

对苯二甲酸对NIH-3T3细胞间隙连接通讯功能的影响

目的 研究对苯二甲酸(TPA)对NIH-3T3细胞间隙连接通讯(GJIC)功能的影响。方法 采用噻唑蓝(MTT)比色法及划痕标记示踪技术(scrape-loading and dye transfer,SLDT),以0、125、250、500、1000、2000μg／ml的剂量染毒细胞,同时设立溶剂对照及阳性对照,观察TPA对GJIC的影响。结果 TPA对细胞GJIC功能抑制随着染毒剂量增大和染毒时间延长而逐渐增强,呈现剂量．效应和时间．效应关系,在250μg／ml或更高剂量染毒1h或更长时间时,细胞GJIC功能被明显抑制,在染毒8h时各剂量组细胞GJIC功能抑制效应均最显著;去除TPA后,GJIC功能有恢复倾向,但在12h内仍不能恢复至正常水平;加入哺乳动物肝脏微粒体酶(S9)能加重TPA对细胞GJIC功能抑制作用。结论 TPA原形及其代谢产物对NIH-3T3细胞GJIC功能均具有抑制作用。     

Synthesis and characterization of triethylene glycol dimethacrylate nanocapsules used in a self-healing bonding resin

Objectives

To date, the production of highly durable dentine bonding is still a challenge. Self-healing bonding resins may provide a new direction for the improvement of the bonding durability. The objective of the current study was to synthesize polyurethane nanocapsules encapsulated with the core material triethylene glycol dimethacrylate (TEGDMA) for use as a major component in a self-healing bonding resin.

Methods

TEGDMA nanocapsules were synthesized via interfacial polycondensation in a miniemulsion, and the TEGDMA nanocapsules were then characterized via Fourier-transform infrared (FTIR) spectrometer, field emission scanning electron microscopy (FESEM), and high-performance liquid chromatography (HPLC) to investigate the morphology, the average TEGDMA loading (DL%), and encapsulation efficiency (EE%). The mechanical property of dental adhesive with different concentrations (0, 3, 6, 9, and 12 wt%) of the TEGDMA nanocapsules were also measured, and the cytotoxicity was investigated using an MTT assay.

Results

FTIR confirmed that the TEGDMA nanocapsules were successfully synthesized. These nanocapsules showed a high drug load. The bond strength of the dental adhesive incorporated with 9 wt% TEGDMA nanocapsules was significantly higher compared with those of the other groups (P < 0.001). Moreover, the biocompatibility of the dental adhesive was not affected by the incorporation of the TEGDMA nanocapsules.

Conclusions

The current study demonstrated the successful synthesis of TEGDMA nanocapsules, and the overall properties of the dental adhesive were not compromised.     

Antitumor activity of extract and isolated compounds from Drechslera rostrata and Eurotium tonophilum

Total extracts of Drechslera rostrata and Eurotium tonophilum in addition of two isolated compounds from their cultures [di-2-ethylhexyl phthalate (H1) and 1,8-Dihydroxy-3-methoxy-6-methyl-anthraquinone (H2)] were tested for their antitumor activity using four human carcinoma cell lines. Antitumor activity was assessed by performing MTT assay to check the % cell viability. The % viability of HCT-116 (colon carcinoma), HeLa (cervical carcinoma), HEp-2 (larynx carcinoma) and HepG-2 (hepatocellular carcinoma) cells decreased after treatment with Drechslera rostrata and Eurotium tonophilum extracts, these effects were ranged from 059.0 ± ?0.1 to 217.0? ± ?0.3?µg/ml on all types of cancer cells. The best activity was recorded for Eurotium tonpholium extract (054.5?±?0.3, 059.0?±?0.5 and 059.0?±?0.1 for HEp-2, Hela, and HepG-2 respectively). The isolated compounds (H1&H2) were found to be responsible about the activities because they recorded the lowest IC₅₀ on tested cell lines with range of 9.5–20.3?μg/ml. Vinblastine sulphate was used as a reference standard and showed in vitro anticancer activity. This study demonstrated that all extracts and isolated compounds have antitumor activity against HCT-116, HeLa, HEp-2 and HepG-2 cells.     

大肠癌细胞和外周血淋巴细胞对化疗药物的体外敏感性

目的:探讨大肠癌细胞及外周血淋巴细胞对化疗药物体外敏感性的及二者的相关性。方法:用MTT法检测40份大肠癌标本及外周血淋巴细胞对氟尿嘧啶(5-FU)、奥沙利铂(L-OHP)、伊立替康(CPT)单药、两药及三药(全量或半量)应用的敏感性。结果:单药最有效的药物依次为伊立替康、奥沙利铂和氟尿嘧啶,敏感率分别为35.0%、27.5%和20.0%;三药联合应用的抑制效果显著优于两药联合(P<0.05),三药全量及半量联合应用效果差异无显著性(P>0.05);癌细胞及外周血淋巴细胞对3种化疗药物的敏感性有很好的相关性(r=0.969)。结论:MTT可用于为大肠癌患者选择合适的化疗药物,由氟尿嘧啶、奥沙利铂及伊立替康的联合应用具有高效协同抑制大肠癌细胞的作用。     

Gallic acid inhibits the growth of HeLa cervical cancer cells via apoptosis and/or necrosis

Gallic acid (GA) is widely distributed in various plants and foods, and its various biological effects have been reported. Here, we evaluated the effects of GA on HeLa cells in relation to cell growth inhibition and death. HeLa cell growth was diminished with an IC₅₀ of approximately 80 μM GA at 24 h whereas an IC₅₀ of GA in human umbilical vein endothelial cells (HUVEC) was approximately 400 μM. GA-induced apoptosis and/or necrosis in HeLa cells and HUVEC, which was accompanied by the loss of mitochondrial membrane potential (MMP; ΔΨ_m). The percents of MMP (ΔΨ_m) loss cells and death cells were lower in HUVEC than HeLa cells. All the tested caspase inhibitors (pan-caspase, caspase-3, -8 or -9 inhibitor) significantly rescued HeLa cells from GA-induced cell death. GA increased reactive oxygen species (ROS) level and GSH (glutathione) depleted cell number in HeLa cells. Caspase inhibitors reduced GSH depleted cell number but not ROS level in GA-treated HeLa cells. In conclusion, GA inhibited the growth of HeLa cells and HUVEC via apoptosis and/or necrosis. The susceptibility of HeLa cells to GA was higher than that of HUVEC. GA-induced HeLa cell death was accompanied by ROS increase and GSH depletion.     

Green approach for the biosynthesis of silver nanoparticles and its antibacterial and antitumor effect against osteoblast MG-63 and breast MCF-7 cancer cell lines

The proposed study aimed to biosynthesize the silver nanoparticles (AgNPs) using aqueous leaf extract of Artocarpus integer (Family-Moraceae) The aqueous extract of A. integer was challenged with silver nitrate solution to produce silver nanoparticles. The color change of the solution confirms the formation of nanoparticles. Characterization of the nanoparticles was performed through UV–Vis spectrophotometry which showed an absorption peak at 425 nm; the FTIR analysis revealed the presence of alkenes, amines, and amides associated with the nanoparticles. Transmission electron microscopy (TEM) revealed the particle size ranges from 5.76 nm to 19.1 nm and is spherical. Thermogravimetric analysis (TGA) showed the actual protein degradation of 9.262% (ΔY) at 71.89 °C. The AgNPs were further evaluated for antibacterial activity on Staphylococcus aureus, Bacillus cereus, Salmonella entertica and Escherichia coli which exhibited good antibacterial activity when compared with ciprofloxacin. Anticancer activity was assessed by MTT assay against osteoblast MG-63 cells which showed IC50 value at 90 μg/ml against MCF-7 cells and 70 μg/ml against MG-63 cells while on normal skin fibroblast showed IC50 value at 110 μg/ml upon 24 h of incubation.     

Ad．RGD—ING4对人鼻咽癌细胞CNE抑癌增效及其机制的研究

目的：研究带RGD的腺病毒介导的ING4对人鼻咽癌细胞CNE生长、凋亡及细胞周期的影响并探讨其可能调节机制。方法：以Ad．RGD-ING4及腺病毒空载体感染CNE细胞,RT—PCR法检测ING4基因的表达水平,Westernblot法检测目的蛋白的表达。用MTT试验检测ING4对CNE细胞生长情况的影响,用AnnexinV—PE／7一AAD双染色测定ING4对CNE细胞凋亡的影响,用PI单染色检测ING4对CNE细胞细胞周期的影响。RT—PCR检测p21、Bcl-2、Bax基因的表达水平的差异,Westernblot法检测Survivin蛋白、Cleaved—caspase3蛋白表达的差异。结果：CNE细胞感染Ad．RGD-ING472h后,ING4在CNE细胞中过表达,CNE细胞的生长受到明显抑制,细胞凋亡率明显升高,G2／M期出现明显阻滞。RT-PCR结果显示,感染Ad．RGD-ING4的CNE细胞中Bcl-2表达下降,p21、Bax表达升高,差异具有统计学意义（均P〈0．01）。Westernblot结果显示Survivin蛋白表达下降,Cleaved—caspase3蛋白表达升高。结论：Ad．RGD-ING4可以对鼻咽癌细胞株CNE的起到抑癌增效作用,这一作用可能是通过下调Bcl-2、Survivin表达及上调p21、Bax、caspase3实现的。