Divergence of Intracellular Trafficking of Sphingosine Kinase 1 and Sphingosine-1-Phosphate Receptor 3 in MCF-7 Breast Cancer Cells and MCF-7-Derived Stem Cell-Enriched Mammospheres

Breast cancer MCF-7 cell-line-derived mammospheres were shown to be enriched in cells with a CD44+/CD24– surface profile, consistent with breast cancer stem cells (BCSC). These BCSC were previously reported to express key sphingolipid signaling effectors, including pro-oncogenic sphingosine kinase 1 (SphK1) and sphingosine-1-phosphate receptor 3 (S1P3). In this study, we explored intracellular trafficking and localization of SphK1 and S1P3 in parental MCF-7 cells, and MCF-7 derived BCSC-enriched mammospheres treated with growth- or apoptosis-stimulating agents. Intracellular trafficking and localization were assessed using confocal microscopy and cell fractionation, while CD44+/CD24- marker status was confirmed by flow cytometry. Mammospheres expressed significantly higher levels of S1P3 compared to parental MCF-7 cells (p < 0.01). Growth-promoting agents (S1P and estrogen) induced SphK1 and S1P3 translocation from cytoplasm to nuclei, which may facilitate the involvement of SphK1 and S1P3 in gene regulation. In contrast, pro-apoptotic cytokine tumor necrosis factor α (TNFα)-treated MCF-7 cells demonstrated increased apoptosis and no nuclear localization of SphK1 and S1P3, suggesting that TNFα can inhibit nuclear translocation of SphK1 and S1P3. TNFα inhibited mammosphere formation and induced S1P3 internalization and degradation. No nuclear translocation of S1P3 was detected in TNFα-stimulated mammospheres. Notably, SphK1 and S1P3 expression and localization were highly heterogenous in mammospheres, suggesting the potential for a large variety of responses. The findings provide further insights into the understanding of sphingolipid signaling and intracellular trafficking in BCs. Our data indicates that the inhibition of SphK1 and S1P3 nuclear translocation represents a novel method to prevent BCSCs proliferation.     

Mutant p53 Mediates Sensitivity to Cancer Treatment Agents in Oesophageal Adenocarcinoma Associated with MicroRNA and SLC7A11 Expression

TP53 gene mutations occur in 70% of oesophageal adenocarcinomas (OACs). Given the central role of p53 in controlling cellular response to therapy we investigated the role of mutant (mut-) p53 and SLC7A11 in a CRISPR-mediated JH-EsoAd1 TP53 knockout model. Response to 2 Gy irradiation, cisplatin, 5-FU, 4-hydroxytamoxifen, and endoxifen was assessed, followed by a TaqMan OpenArray qPCR screening for differences in miRNA expression. Knockout of mut-p53 resulted in increased chemo- and radioresistance (2 Gy survival fraction: 38% vs. 56%, p < 0.0001) and in altered miRNA expression levels. Target mRNA pathways analyses indicated several potential mechanisms of treatment resistance. SLC7A11 knockdown restored radiosensitivity (2 Gy SF: 46% vs. 73%; p = 0.0239), possibly via enhanced sensitivity to oxidative stress. Pathway analysis of the mRNA targets of differentially expressed miRNAs indicated potential involvement in several pathways associated with apoptosis, ribosomes, and p53 signaling pathways. The data suggest that mut-p53 in JH-EsoAd1, despite being classified as non-functional, has some function related to radio- and chemoresistance. The results also highlight the important role of SLC7A11 in cancer metabolism and redox balance and the influence of p53 on these processes. Inhibition of the SLC7A11-glutathione axis may represent a promising approach to overcome resistance associated with mut-p53.     

Molecular Mechanisms of the Deregulation of Muscle Contraction Induced by the R90P Mutation in Tpm3.12 and the Weakening of This Effect by BDM and W7

Point mutations in the genes encoding the skeletal muscle isoforms of tropomyosin can cause a range of muscle diseases. The amino acid substitution of Arg for Pro residue in the 90th position (R90P) in γ-tropomyosin (Tpm3.12) is associated with congenital fiber type disproportion and muscle weakness. The molecular mechanisms underlying muscle dysfunction in this disease remain unclear. Here, we observed that this mutation causes an abnormally high Ca²⁺-sensitivity of myofilaments in vitro and in muscle fibers. To determine the critical conformational changes that myosin, actin, and tropomyosin undergo during the ATPase cycle and the alterations in these changes caused by R90P replacement in Tpm3.12, we used polarized fluorimetry. It was shown that the R90P mutation inhibits the ability of tropomyosin to shift towards the outer domains of actin, which is accompanied by the almost complete depression of troponin’s ability to switch actin monomers off and to reduce the amount of the myosin heads weakly bound to F-actin at a low Ca²⁺. These changes in the behavior of tropomyosin and the troponin–tropomyosin complex, as well as in the balance of strongly and weakly bound myosin heads in the ATPase cycle may underlie the occurrence of both abnormally high Ca²⁺-sensitivity and muscle weakness. BDM, an inhibitor of myosin ATPase activity, and W7, a troponin C antagonist, restore the ability of tropomyosin for Ca²⁺-dependent movement and the ability of the troponin–tropomyosin complex to switch actin monomers off, demonstrating a weakening of the damaging effect of the R90P mutation on muscle contractility.     

LabVIEW 7 Express平台上的液压测试系统

为了满足工程实践对先进液压测试技术的需要，研究了基于美国NI公司最新推出的LabVIEW 7 Express虚拟仪器开发平台的液压测试系统的软硬件结构，介绍了测试系统实现网络通讯、数据库链接和多种编程语言联合作业的方法，提出了现代液压测试系统的技术方案。     

STUDY OF INTERFACES AND SURFACES OF YBa_2Cu_3O_(7-x)-Ag

Interfaces and surfaces of YBa_2Cu_3O_(7-x)(YBCO)-Ag have been studied by SEM-EDXand AES.No effect of Ag on 123 structure in X-ray diffraction pattern was observedfor 0.4 mol Ag doped YBCO.AES analysis indicated that Ag segregated on surfaceof YBCO and resulted in decrease of YBCO-metal lead resistance.In addition,solutionand segregation of Ag as elemental state were often appeared on interfaces and surfacesof high temperature annealed YBCO,whether elemental Ag or compound Ag_2O andAgNO_3 adopted as doping material.     

LD7铝合金热变形行为的实验研究

在G1eeble-1500热模拟试验机上对LD7铝合金试样在变形程度为60％，变形温度为360～480℃、变形速率为0．0l～1s^-1的条件下进行等温压缩试验，然后对其进行固溶处理。根据试验结果分析热变形参数对合金流动应力和固溶之后显微组织的影响，可以得到如下结论：LD7铝合金是动态回复型合金，合金的流动应力随温度的升高而降低，随应变速率的升高而升高；合金的晶粒尺寸随温度的升高而增大，随变形程度和变形速率的增大而减小。     

LD7铝合金时效工艺的研究

通过试验系统地研究了ＬＤ７铝合金的单级和两级时效工艺。结果表明，与传统的单级时效相比，两级时效在不降低室温强度，硬度的前提下，可显著提高合金的电导率及高温性能。     

用相变方法制取高纯度超细钽粉的研究

报导了通过相变热处理使Ｋ２ＴａＦ７晶体参数发生变化，以利于制取高纯度超细钽粉的实验依据和结果。结果表明，Ｋ２ＴａＦ７在２００～２５０℃发生可逆相变；相变后Ｋ２ＴａＦ７晶体的粒径可由原来的１３５１．８μｍ减小至７０．８μｍ，极大地提高了比表面积；相变后，杂质极大挥发，提高了Ｋ２ＴａＦ７的纯度。     

稀土Y对纳米粉90W-Ni-Fe合金性能和微观结构的影响

采用SEM和金相仪器对添加不同Y量的纳米粉90W-7Ni-3Fe合金试样断口进行形貌观察和W晶粒测试：并对烧结态试样的相对密度、抗拉强度、延伸率等性能进行测定与分析。结果表明：当Y量在0．2％～0．6％（质量百分数，下同）时，试样相对密度为99.2％～99．6％，抗拉强度为1000MPa-1100MPa，延伸率为16％-19．5％：添加0．4％Y后，W晶粒从原来的20μm～25μm减小到12μm，W晶粒形状由不加稀土时的球形变为近球形或多边形。添加0．4％Y后，在界面上形成了W13．07Ni2.96Fe1.52Y23．65Ox（摩尔数比）的中间相，其阻止了W原子在粘结相中的扩散，阻止了W晶粒长大，细化了W晶粒，提高了合金的性能。