Initial study on apoptosis in HepG-2 Human heptocarcinoma cell line by CSS

Objective To discuss on mechanism of the killing and apoptosis inducing effect induced by total alkaloid in the CSS(Capparis spinosa L.saponin,CSS)on human hepatocarcinoma cell Line HepG-2.Methods The killing effect of the CSS on human hepatocarcinoma cell Line HepG-2 was observed by MTT method.Morphological observation of the HepG-2 cells was completed by fluorescence microscope.This test was signed to observe the changes of the cell cycle of HepG-2 cells affected by the CSS by PI single-staining,and to observe if there were typical apoptosis peaks.The apoptosis inducing effect and changing of mitochondria membrane potential of the CSS on the HepG-2 cells were studied by flow cytometry.The effect of intracellular Ca2+ level of CSS on the HepG-2 cells was measured by laser confocal microscope.Results CSS has growth inhibiting on the HepG-2 and seems to be enhanced with the increasing concentration of CSS,and its IC50 value was 46.16 μg·mL-1.The HepG-2 cells are characteristic apoptosis morphologic changed,and the apoptosis percentage is increased to 66.652% in the 50 μg·mL-1 dosage group.The cells cycle has been changed obviously that the progresses of cells cycle of G1 period and G2 period in high dosage group have been blocked,and the cellular proportion in G2 period is decreased by the function of CSS for 24 h.The mitochondria membrane potential of HepG-2 cells induced by CSS is decreased in various degrees.In addition,the intracellular Ca2+ level is increased by the function of CSS in the middle and high dose groups.Conclusions The CSS has obviously killing and apoptosis inducing effect on human hepatocarcinoma cell Line HepG-2 by the mechanism of decreasing the mitochondria membrane potential and increasing the intracellular Ca2+ level.

Study on preliminary mechanism of apoptosis in HepG-2 by CSA

Objective To study on the mechanism of killing and apoptosis inducing effect of total alkaloid in the CSA(Capparis spinosa L.alkaloid,CSA)on human hepatocarcinoma cell Line HepG-2.Methods The killing effect of the CSA on human hepatocarcinoma cell Line HepG-2 was measured by MTT method.Morphological observation of the HepG-2 cells was completed by fluorescence microscope.The apoptosis inducing effect and changing of mitochondria membrane potential of the CSA on the HepG-2 cells were measured by flow cytometry.In addition,effect of intracellular Ca2+ level of the CSA on the HepG-2 cells was studied by laser confocal microscope.Results The CSA has obvious cytotoxicity on the HepG-2 and seems to be dose-dependent,and its IC50 value is 162.4 μg·mL-1.The HepG-2 cells have characteristic morphologic changes of apoptosis by the function of CSA,and the apoptosis percentage is higher than the natural one.The progress of cells cycle from S phase to G2 phase has been blocked,and the mitochondria membrane potential is markedly decreased,and the intracellular Ca2+ level is increased by the function of CSA.Conclusions The CSA has obviously killing and apoptosis inducing effect on human hepatocarcinoma cell Line HepG-2 by the mechanism of decreasing the mitochondria membrane potential and increasing the intracellular Ca2+ level.

Effect on apoptosis、mitochondrial membrane potential and Ca^(2+) concentration in HepG-2 by CSEO

Objective To study on the mechanism of growth inhibiting and apoptosis inducing effect of total alkaloid in the CSEO(Capparis spinosa L.essential oil,CSEO)on human hepatocarcinoma cell Line HepG-2.Methods The growth inhibiting effect of the CSEO on human hepatocarcinoma cell Line HepG-2 was measured by MTT method.Morphological observation of the HepG-2 cells was completed by fluorescence microscope.The changing of mitochondrion membrane potential induced by CSEO was observed by staining with Rhodamine123.Effect of the CSEO on intracellular Ca2+ level of the HepG-2 cells was measured by laser confocal microscope.Results The CESO has obvious growth inhibiting effect on the HepG-2 and seems to be dose-dependent,and its IC50 is 127.5 μg·mL-1.The characteristic apoptosis morpha of HepG-2 cells has been observed,and the apoptosis percentage increase to 44.447% in the 300 μg·mL-1 dosage group.In addition,the progress of cells cycle of G1 period has been blocked,and the cellular proportion in S and G2 period is decreased in the 75 μg·mL-1 and 150 μg·mL-1 dosage groups by the function of CSEO for 48 h.The mitochondria membrane potential(Δψm)effected by CESO is decreased,while the curve moves toward left.In addition,the intracellular Ca2+ level is increased by the function of CESO in the middle and high dose groups.Conclusions The CESO has obviously growth inhibiting and apoptosis inducing effect on human hepatocarcinoma cell Line HepG-2 by the mechanism of decreasing the mitochondria membrane potential and increasing the intracellular Ca2+ level.

The expression of Lin28B was co-regulated by H3K4me2 and Wnt5a/β-catenin/TCF7L2

Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells. The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited. In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed. DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured. Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells. Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp. The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis. The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated. Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling. Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area. In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression. H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B. In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression. Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.

IGF1和IGF2表达与非小细胞肺癌淋巴结转移的相关性

目的研究胰岛素样生长因子1和2(IGF1、IGF2,合称为IGFs)在非小细胞肺癌(NSCLC)组织中的表达,探讨其与NSCLC区域淋巴结转移的关系。方法采用免疫组织化学和免疫印迹技术检测IGF1和IGF2在65例NSCLC、29例癌旁肺组织和19例良性肺病变组织中的表达。结果IGF1和IGF2在NSCLC中的表达率显著高于良性肺组织。IGF1和IGF2在伴有区域淋巴结转移肺癌中的表达显著高于无淋巴结转移肺癌组织。IGF1和IGF2在伴有N2或≥5个转移淋巴结肺癌组织中的表达显著高于在仅伴有N1或<5个转移淋巴结肺癌组织。IGF2在伴有多组转移淋巴结肺癌组织中的表达显著高于仅有一组转移淋巴结肺癌组织。结论IGFs不仅在NSCLC中过表达,而且与区域淋巴结转移的部位、数量及组数相关联,提示IGFs在NSCLC发生和发展中可能起重要作用,IGFs为NSCLC生物学行为尤其是淋巴结转移的判断提供一个新的有意义的指标。

Effects of suppressing glucose transporter-1 by an antisense oligodeoxynucleotide on the growth of human hepatocellular carcinoma cells

BACKGROUND:The glucose transporter-1(Glut-1),a key ratelimiting factor in the transport and metabolism of glucose in cancer cells,is over-expressed in many human cancer cells and this overexpression is correlated with poor biological behavior. The increased levels of Glut-1 expression in hepatocellular carcinoma(HCC)cells functionally affect tumorigenicity.This study was undertaken to investigate effects of suppressing Glut-1 by an antisense oligodeoxynucleotide(AS-ODN)on the growth of human hepatocellular carcinoma(HepG-2)cells. METHODS:We used AS-ODN targeting against the Glut-1 gene in a HepG-2 cell line.There were four experimental groups: empty pcDNA3.1 vector(mock transfection),pcDNA3.1-anti-Glut(+),pcDNA3.1-Glut(+),and non-transfected HepG-2 cells. The Glut-1 mRNA expression was detected by RT-PCR and the Glut-1 protein expression by Western blotting after cell culture, and the glucose uptake was detected after glucose stimulation in each group. RESULTS:Compared with non-transfected HepG-2 or Glut-1 pcDNA3.1,a down-regulation of Glut-1 mRNA in HepG-2 cells transfected with anti-Glut-1 pcDNA3.1 was noted(P<0.05).Glut-1 protein in HepG-2 cells transfected with Glut-1 AS-ODN was decreased compared with non-transfected HepG-2,Glut-1 pcDNA3.1,or empty vectors. Glucose uptake by the HepG-2 cells transfected with AS-ODN was decreased at 1 hour after glucose stimulation.CONCLUSIONS:The application of Glut-1 AS-ODN can down-regulate the expression of Glut-1 at mRNA and protein,and inhibit glucose uptake partially in HepG-2 cells.The Glut-1 gene maybe a potential therapeutic target for HCC.

咖啡酸对硝基苯乙酯对肝癌HepG-2细胞株裸鼠增殖瘤的抑制作用

通过用肝癌HepG-2细胞株建立BALB/c裸鼠移植瘤模型,观察3、6、12 mg/kg剂量的CAPE-NO_2对裸鼠移植瘤的抑制率,并检测了小鼠胸腺和脾脏指数、腹腔巨噬细胞吞噬作用、脾脏T淋巴细胞增殖反应、NK细胞活性、血清NO及IL-1、TNF-α含量。结果,3、6、12 mg/kg剂量的CAPE-NO_2对裸鼠移植瘤的抑制率分别为50.67%、55.58%和71.31%;与模型组相比,CAPE-NO_2组脾脏指数、胸腺指数、腹腔巨噬细胞吞噬作用、脾脏T淋巴细胞增殖反应、NK细胞活性、IL-1的水平升高(P<0.01),而血清NO、TNF-α水平降低(P<0.01),呈剂量依赖效应。上述结果表明,CAPE-NO_2对肝脏HepG-2细胞株荷瘤小鼠移植瘤有很好的抑制作用,与其能提高荷瘤小鼠的免疫功能密切相关。

增塑盐改性准固态电解质染料敏化纳米晶太阳能电池

实验使用一种增塑性试剂—双（三氟甲基磺酰）亚胺锂改性PEO-PVDF基聚合物电解质设计并制备了一系列不同浓度双（三氟甲基磺酰）亚胺锂[LiN（SO2CF3）2,LiTFSI]改性的PEO基聚合物电解质。在聚合物电解质中LiTFSI起着增塑剂的作用,经其改性后,聚合物电解质的玻璃化转变温度降低,有利于高分子链段运动和离子传输,进而提高离子的电导率。最终对经增塑盐改性的电解质的特性及其所组装的染料敏化纳米晶太阳电池的光电转换性能进行了研究。

B细胞淋巴瘤／白血病-2相关X蛋白和细胞周期蛋白依赖性激酶抑制蛋白27在中心型软骨肉瘤组织的表达及临床意义

目的研究中心型软骨肉瘤（Central chondrosarcoma）中B细胞淋巴瘤／白血病12相关X蛋白（bax）及细胞周期蛋白依赖性激酶抑制蛋白27（p27^kipl）蛋白的表达与病理分级及预后的关系，并探讨两者表达的临床意义。方法选取60例确诊的中心型软骨肉瘤患者，术后随访超过5年，按照病理分型及预后对其病理标本分别进行分组，并选用20例非肿瘤软骨标本作为对照，采用免疫组织化学法检测上述标本中bax和p27^kipl蛋白的表达。结果60例中心型软骨肉瘤中，bax蛋白各级别间阳性表达率分别为Ⅰ级90％、Ⅱ级78．9％、Ⅲ级66．7％，差异有统计学意义（P〈0．05），在不同预后组中bax蛋白阳性表达率分别为治愈组80．0％、复发组75．0％、转移组75．0％，差异无统计学意义（P〉0．05）；p27^kipl州蛋白各级别阳性表达率分别为Ⅰ级90％、Ⅱ级70．9％、Ⅲ级61．9％，差异有统计学意义（P〈0．05），在不同预后分组中p27^kipl蛋白阳性表达率分别为治愈组80．4％、复发组66．7％、转移组37．5％，差异亦有统计学意义（P〈0．05）。结论bax及p27^kipl蛋白的表达与中心型软骨肉瘤的病理分级明显相关，但bax不能作为判断预后的指标，而p27^kipl蛋白则可作为判断预后的指标。