mTOR kinase inhibitor CC-223 suppresses human breast cancer cell viability

AIM: To investigate the inhibitory effect of CC-223, an inhibitor of mammalian target of rapamycin (mTOR) kinase, on the viability of human breast cancer cells and its mechanism. METHODS: The inhibitory effect of CC-223 on the viability of MCF-7 cells and MDA-MB-231 cells was measured by CCK-8 assay. The cell cycle distribution of breast cancer cells was examined by flow cytometry. The expression of cell cycle-related proteins and oncoproteins c-Myc and survivin was analyzed by Western blot. RESULTS: CC-223 significantly inhibited the viability of both MCF-7 and MDA-MB-231 cells (P<0.05). CC-223 induced cell cycle arrest in both G₁ phase and G₂/M phase in the MCF-7 cells (P<0.05). However, low concentration of CC-223 treatment resulted in the accumulation of MDA-MB-231 cell cycle in the G₂/M phase, and the cell number in G₁ phase was unaffected. Treatment with CC-223 for 24 h clearly inhibited the protein levels of cyclin B1, cyclin D1 and phosphorylated cell division cycle protein 2 in the breast cancer cells (P<0.05). CC-223 suppressed the expression of c-Myc and survivin in both MCF-7 cells and MDA-MB-231 cells (P<0.05). CONCLUSION: CC-223 inhibits cell viability by blocking cell cycle progression and down-regulating expression of c-Myc and survivin in both MCF-7 and MDA-MB-231 cells.     

Expression and function of circular RNA_0000231 in non-small-cell lung cancer

AIM: To investigate the expression and function of circular RNA_0000231 (circ_0000231) in non-small-cell lung cancer (NSCLC). METHODS: RT-qPCR was used to detect the expression of circ_0000231 in the NSCLC tissues and cell lines. circ_0000231 small interfering RNA (si-circ_0000231) or negative control siRNA of circ_0000231 (NC) was transfected into the NSCLC cells. The proliferation and apoptosis of the NSCLC cells were detected by CCK-8 assay, colony formation assay and flow cytometry, respectively. The expression of cyclin D1 (CCND1) and anti-apoptotic protein Bcl-2 were determined by RT-qPCR and Western blot. RESULTS: The expression of circ_0000231 in the NSCLC tissues and cell lines was significantly up-regulated compared with precancerous tissues and lung epithelial cells BEAS-2B (P<0.05). After transfection of NSCLC cells with si-circ_0000231, the cell viability, colony formation numbers were significantly decreased, and the apoptotic rate in si-circ_0000231 group was significantly increased as compared with NC group (P<0.01). In addition, the results of RT-qPCR and Western blot showed that transfection of si-circ_0000231 inhibited the expression of CCND1 and Bcl-2 (P<0.01). CONCLUSION: The expression of circ_0000231 is significantly increased in the NSCLC tissues and cells. Knock-down of circ_0000231 expression significantly inhibits the proliferation of NSCLC cells.     

尼罗罗非鱼精巢支持细胞分离培养体系建立及优化

本研究分离培养尼罗罗非鱼(Oreochromis niloticus)精巢支持细胞(Sertoli cells, SCs)，建立并优化尼罗罗非鱼精巢支持细胞分离培养体系。无菌条件下，取发育至第Ⅲ期的尼罗罗非鱼精巢，PBS清洗后，剪碎精巢组织，0.25%胰蛋白酶消化，用含10%犊牛血清(Newborn bovine serum, NBS)的L-15培养液终止消化，过滤、离心，获得细胞。差速贴壁法获得SCs后，在26℃、无CO2饱和湿度恒温培养箱中培养。分别采用含10% NBS的L-15、M199、F12培养液，含5%、10%、15% NBS的L-15培养液，含1%罗非鱼血清的L-15+10% NBS培养液培养尼罗罗非鱼SCs。各组每2 d取细胞计数，绘制SCs生长曲线；甲基绿染色，倒置显微镜下观察SCs的形态。尼罗罗非鱼SCs培养1~2 d，细胞贴壁；培养3~5 d，细胞完全贴壁并迅速增殖。单个SCs呈不规则多边形，其核位于胞质中央，呈卵圆形，胞质中可见吞噬颗粒和空泡，空泡聚集于支持细胞胞质的两极或散布于核的四周。SCs在L-15培养液中贴壁生长，在F12、M199培养基中较难贴壁。与F12、M199培养液相比，L-15培养液更有助于尼罗罗非鱼SCs生长增殖(P<0.01)。与添加5%和15% NBS相比，在L-15培养基中添加10% NBS更有助于尼罗罗非鱼SCs生长增殖(P<0.05)。与未添加尼罗罗非鱼血清相比，在10% NBS+L-15培养中添加1%尼罗罗非鱼血清，能显著促进SCs生长增殖(P<0.05)。研究表明，采用胰酶消化差速贴壁法获得尼罗罗非鱼精巢支持细胞，10% NBS+1%罗非鱼血清+L-15培养液培养，能显著促进尼罗罗非鱼精巢支持细胞生长增殖。     

甲状腺素对猪小肠上皮细胞miR-let-7a基因表达及细胞增殖的影响

miR-let-7a在动物细胞的分化、增殖与凋亡等方面发挥越来越重要的作用。甲状腺激素(TH)作用非常广泛,机体的每个细胞几乎都是TH作用的靶细胞,其可以促进组织分化、生长和成熟。本实验用甲状腺素(T4)浓度分别为(0、0.02、0.03、0.05、0.075、0.1、0.2μmol/L)在体外培养猪的小肠上皮细胞。结果表明:T4处理组的细胞体积形态相对于空白对照组没有明显变化;当T4添加浓度为0.03μmol/L时,细胞的增殖率显著低于其他组(P0.05);当T4浓度为0~0.03μmol/L时,let-7a的表达随着添加剂量的增加而升高,浓度从0.03~0.2μmol/L变化时,let-7a的表达呈现降低趋势,浓度为0.03μmol/L时表达量极显著高于其他组(P0.01)。let-7a的表达量与细胞增殖呈负相关。     

Identification and establishment of sorting method for isolating CD11b+ myeloid cells in human hepatic metastases from colorectal cancer

AIM: To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b⁺ myeloid cells in hepatic metastases from colorectal cancer.METHODS: Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion, and then the CD11b⁺ myeloid cells were isolated by flow cytometry. The sorted cells were identified by immunocytochemistry. The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining. The cell purity was evaluated by flow cytometry.RESULTS: Sufficient numbers of CD11b⁺ cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion. The purity of the cells was confirmed by statistical analysis (P<0.05). The positive rates of the cells before and after sorting were significantly different (P<0.01). The positive cells were verified by immunocytochemical method. Meanwhile, the sorted cells had complete morphology and good activity.CONCLUSION: The CD11b⁺ myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity, good activity and complete morphology.     

低溶胀壳聚糖/丝素蛋白复合膜的制备及性能测试

为改善单纯壳聚糖膜用作医用创面敷料在湿环境下的力学性能,将壳聚糖(CS)和家蚕丝素蛋白(SF)混合,采用延流法制备壳聚糖/丝素蛋白(CS/SF)复合膜。物理与机械性能测试表明:CS/SF复合膜的溶胀率较单纯CS膜显著降低,且其溶胀能力具有pH依赖性;CS/SF复合膜的水蒸气透过率较单纯CS膜略有下降,但仍保持在2 043～3 363 g/(m2.d);CS/SF复合膜在湿环境下的力学强度较单纯CS膜显著提高,当加入SF的质量分数为15%时,复合膜的力学性能最好,其拉伸强度为34.5 MPa±1.8 MPa,比单纯CS膜提高近18%,而其断裂伸长率也由初始的83.1%±6.7%提高到119.3%±8.2%。CS/SF复合膜具有较好的药物释放性能,对抗感染药物盐酸万古霉素表现出在初始1 h内的突释行为和随后较长时期的缓释行为,十分有利于降低感染和加速伤口愈合。此外,CS/SF复合膜对STO小鼠胚成纤维细胞具有良好的生物相容性。各项性能测试显示CS/SF复合膜作为医用敷料具有潜在的应用前景。     

丛枝菌根真菌和糖醇螯合钙对草莓采后果实硬度和细胞壁酶活及其关键基因表达的影响

以盆栽‘红颜’草莓为试材，研究了摩西管柄囊霉（Funneliformis mosseae）和糖醇螯合钙不同浓度（0.10%、0.15%、0.20%）处理对草莓采后果实硬度和细胞壁酶活的变化，同时，初步探究了不同处理对草莓果实软化关键基因FaPG1、FaβGal4的表达水平的影响。结果表明，与对照相比，以糖醇螯合钙单独施用（0.15%和0.20%）、接种丛枝菌根真菌（Arbuscular mycorrhizal fungi, AMF）配施糖醇螯合钙均能显著提高草莓果实硬度。同时，不同处理均抑制多聚半乳糖醛酸酶（PG）、果胶甲酯酶（PME）、β-半乳糖苷酶（β-Gal）活性的上升，草莓果实软化关键基因FaPG1和FaβGal4在处理后均下调表达。在接种AMF条件下，随着糖醇螯合钙浓度的增加，果实硬度逐渐增强，其中以接种AMF联合施用0.20%糖醇螯合钙溶液效果最好。     

Cell direct reprogramming：a new technique for treating diabetes

Diabetes is characterized by an absolute or relative deficiency in β-cell mass, which cannot be reversed with existing therapeutic strategies. The restoration of the endogenous islet β-cells can stabilize the level of blood glucose. The islet β-cells can be obtained from the directional differentiation of stem cells, but the process is complex and has the risk of teratomas generation. Cell direct reprogramming, one terminal differentiated cell can transdifferentiate into another kind of terminal differentiated cell, which is other than directional differentiation from stem cells. Direct reprogramming gives rise to the generation of islet β-cells from one terminal differentiated cell, may be preferable for diabetes therapy because of its unique advantage.     

Inhibitory Effects of AURKB Gene on Apoptosis and Cancer Cell Growth in HCT 116 Cells

[Objectives]To explore the inhibitory effect of AURKB gene in apoptosis and cancer cell growth in HCT 116 cells.[Methods]The in vitro cytology studies were carr...