当归红芪超滤物对重离子12C6+辐射肝癌细胞DNA损伤修复的影响

将人肝癌H22细胞分成4组,分别为对照组、药物组(100 mg/L)、辐射组(2 Gy)及联合组(100 mg/L药物 + 2 Gy照射),采用CCK-8法、单细胞凝胶电泳、γ-H2AX免疫荧光原位杂交技术以及Western Blotting印迹法,研究当归红芪超滤物(Radix Angelicae Sinensis and Radix Hedysari, RAS-RH)对重离子¹²C⁶⁺辐射引起人肝癌H22细胞DNA损伤修复的影响和其可能的机制。结果表明,在0～72 h和给药剂量为5～200 mg/L范围内,RAS-RH对人肝癌H22细胞的增殖抑制作用具有时间和剂量依赖性,其20%抑制浓度IC₂₀为(117.6±2.15)mg/L;单细胞凝胶电泳显示联合组头部DNA含量低于辐射组,而尾部DNA含量、尾距TM、Olive尾距OTM均高于辐射组;γ-H2AX免疫荧光原位杂交技术发现RAS-RH不增加重离子¹²C⁶⁺辐射引起的DNA损伤,但在2—12 h,DNA双链断裂的γ-H2AX foci修复作用被RAS-RH抑制,DNA损伤持续存在;Western Blotting显示RAS-RH通过下调Ku70/80及Rad51的蛋白表达,抑制γ-H2AX的聚集。以上结果说明RAS-RH对人肝癌H22细胞的辐射增敏作用可能是下调DNA损伤修复相关因子Ku70/80及Rad51的表达。     

The anti-tumor effects of cordycepin-loaded liposomes on the growth of hepatoma 22 tumors in mice and human hepatoma BEL-7402 cells in culture

Liposomes have successfully been used for decades to encapsulate and protect drugs that are prone to deactivation in the body. The present study aimed to demonstrate the use of liposomes to encapsulate cordycepin, an adenosine analog that quickly loses its activity in vivo. The cordycepin-loaded liposomes were prepared by the ammonium sulfate gradient approach, and its in vitro and in vivo antitumour activities were evaluated using BEL-7402 cells and hepatocellular carcinoma H22 transplanted tumors, respectively. An MTT assay was used to observe the cytotoxicity of cells treated with cordycepin and cordycepin-loaded liposomes in vitro. High-content screening (HSC) was carried out using Hoechst 33342 to detect apoptotic cells and the ratio of cells in different cell cycle stages. The data demonstrated that both the cordycepin and the cordycepin-loaded liposomes resulted in clear cytotoxicity with IC50 values of 18.97 and 29.39?μg/mL, respectively. The latter showed significantly strong inhibitory effects on H22 tumor growth in mice, while the former did not show any inhibitory effects on tumor growth. In addition, the HSC assay showed that the cordycepin-loaded liposomes resulted in a higher rate of apoptosis than the cordycepin alone in BEL-7402 cells. Further data analysis revealed that the cells treated with cordycepin-loaded liposomes were predominately arrested at the G2/M phase (p?p?in vivo anti-tumor activity of cordycepin.     

介电电泳细胞分析芯片结构设计及富集效率优化

为了研制高富集效率的介电电泳细胞分析芯片,首先从介电电泳力出发,推导了悬浮细胞所受的介电电泳力公式。通过对比常规微电极的电场强度分布,选择叉指式阵列微电极构建介电电泳芯片;通过模拟不同结构参数下微通道中的电场分布对芯片结构参数进行优化设计。针对Hep G2肝癌细胞,分别分析了细胞受介电电泳力、流体力以及重力作用下的运动情况,获得了Hep G2肝癌细胞富集的初步优化条件。为了对模拟结果进行验证,采用微加工技术制作了介电电泳细胞分析芯片。以Hep G2肝癌细胞为待测样品,当芯片所施加正弦交流电压为5 V,频率为4 MHz时,获得了88.89%的富集效率。     

华蟾素注射液对人肝癌HepG-2细胞增殖及周期的影响

Objective: The aim of our study was to investigate the effect of Cinobufacini injection on the proliferation and cell cycle of human hepatoma HepG-2 cells. Methods: Cell proliferation was assessed by MTT assay, cell cycle distributionwas detected by the flow cytometry (FCM). The expression of Cyclin A, CDK2 mRNA levels were examined by RT-PCR.Quantitative colorimetric assay was used to analyze Cyclin NCDK2 activity in HepG-2 cells. Results: Cinobufacini injection significantly inhibited HepG-2 cells proliferation in dose- and time-dependent ways; FCM analysis showed Cinobufacini injection induced cell cycle arrest at S phase; RT-PCR assay showed Cinobufacini injection down-regulated Cyclin A, CDK2expression at mRNA levels; Quantitative colorimetric assay showed Cinobufacini injection deceased Cyclin A/CDK2 activity in HepG-2 cells. Conclusion: Cinobufacini injection can inhibit human hepatoma HepG-2 cells growth, induce cell apoptosis and induce cell cycle arrest at S phase, the mechanism of which might be partly related to the down-regulation of Cyclin A,CDK2 mRNA expression and inhibition of Cyclin A/CDK2 activity.     

Selective anti-hepatoma treated with titanium oxide nanoparticles in vitro

Effects of titanium oxide (TiO₂) nanoparticles on Bel-7402 human hepatoma hepatoma cells and L-02 human hepatocytes at different times were observed. Using cell culture, cell growth curves of Bel-7402 cells and L-02 cells treated with TiO₂ nanoparticles were examined by MTT assay, and the cellular ultrastructure was observed by an analytical transmission electron microscope (ATEM). It is found that OD value of Bel-7402 cell treated with TiO₂ nanoparticles for 48–144h is obviously lower than that of control group (p<0.01). However the growth curve of L-02 cells is almost not affected by TiO₂ nanoparticles. ATEM and energy dispersive X-ray (EDX) analyses show that there are obvious vacuoles increased heterolysosome, and particles with high electron density which are confirmed to be TiO₂ nanoparticles in Bel-7402 cytoplasm. More interestingly, it is alse found that TiO₂ nanoparticle obviously inhibits the proliferation of hepatoma cells by altering lysosome activity and destroying cytoplasm structure. The inhibition on proliferation of hepatocytes by TiO₂ nanoparticles is much slighter. The results demonstrate that TiO₂ nanoparticle has different killing effects on cancer cell and normal cell. CAO Xian-ying: Born in 1962 Funded by the National Natural Science Foundation of China (No:39770225)     

The 4‐acetylantroquinonol B isolated from mycelium of Antrodia cinnamomea inhibits proliferation of hepatoma cells

BACKGROUND: Antrodia cinnamomea is known for its antihepatoma activity, yet the identity of its active compound was unclear. In this study, a 5‐ton fermenter was used to prepare sufficient mycelium of A. cinnamomea for active compound isolation and identification. RESULTS: Using antiproliferative activity toward HepG2 cells as guidance in the isolation process, 4‐acetylantroquinonol B was purified and identified to be the major bioactive compound of A. cinnamomea cultivated by submerged fermentation. The median effective doses (EC₅₀) of 4‐acetylantroquinonol B for HepG2 cells were 0.10 ± 0.00 and 0.08 ± 0.00 µg mL^?1 for 72 and 96 h treatments, respectively. The selective indices of 4‐acetylantroquinonol B were 100 and 125 for 72 and 96 h treatments, respectively, indicating that this compound had high selective activity for hepatoma cells. CONCLUSION: 4‐Acetylantroquinonol B is the major antihepatoma constituent of Antrodia cinnamomea mycelium produced by submerged fermentation. Copyright © 2010 Society of Chemical Industry     

抗癌药物筛选的新方法--紫杉醇药效的压电细胞芯片研究初探

以人肝癌细胞株HepG2为研究对象,以压电细胞芯片技术为研究手段,进行了紫杉醇药效的压电细胞芯片实时检测;同时与传统的MTT比色方法进行对比,得到了较为一致的实验结果,即药物浓度与细胞的生长抑制率呈正相关.与传统方法相比,压电细胞芯片技术更省时,更动态,是一种体外筛选药物的好方法.     

小黑药亲电成分干预混合游离脂肪酸诱导的人肝癌细胞株HepG2脂肪变性研究

采用GSH功能化磁珠靶向敲出小黑药亲电成分并用LC-MS表征,基于游离脂肪酸(FFA)诱导的HepG2细胞脂肪变性模型,评价小黑药亲电成分降低肝细胞脂质累积和氧化应激的作用和机制。结果表明:小黑药亲电成分主要集中在乙酸乙酯部位;在0.5~2.0mg/mL浓度下,小黑药石油醚部位、乙酸乙酯部位、水部位均具有降低脂肪变性细胞脂质累积和ROS生成的活性,其中乙酸乙酯部位在各浓度下效果最好;乙酸乙酯部位在各浓度下具有降低细胞内TG、TC水平和提高细胞内抗氧化酶活性的作用;经过GSH功能化磁珠靶向敲出亲电化合物后,乙酸乙酯部位在2.0μg/mL浓度下对细胞内TG水平的降低和细胞内抗氧化酶水平的提高效果显著减弱。乙酸乙酯部位萃取物上调Nrf2及下游NQO1、HO-1、GCLC基因,下调脂质合成基因SREBP1c、ACC1、FAS的表达,促进脂质分解基因PPARα、CPT1A的表达;而靶向敲出亲电成分后,乙酸乙酯部位萃取物调控脂代谢和抗氧化相关基因的作用明显下降。LC-MS表征亲电成分敲出前后乙酸乙酯部位样品,发现了7个变化的主要质谱峰,推测其为主要的亲电化合物。小黑药中亲电化合物可以改善脂肪酸诱导的脂肪变性,其机制主要与抗氧化和脂代谢相关基因的调节有关。