蛋白质组学在肝癌研究中的应用

肝癌是世界上流行最广的恶性肿瘤之一,但发病机制至今仍不明了,且缺乏有效的早期诊断方法和治疗靶点。随着人类基因组计划的实施和推进,生命科学研究已进入了后基因组时代。研究的重点从基因组学转向蛋白质组学(proteomics)。蛋白质组学将成为研究肿瘤及相关疾病的最有效方法之一。本文着重就蛋白质组学在肝癌研究的应用作一综述。     

Analysis of the in vitro synergistic effect of 5-fluorouracil and cisplatin on cervical carcinoma cells

5-Fluorouracil (5-FU) is currently being used as an anticancer drug to reduce tumor bulk in order to increase the operability rate and postoperative survival in patients with cervical cancer, which has been combined with cisplatin (CP) because of its superior activities observed in human carcinoma cells. However, the combined anticancer effect of 5-FU and CP in cervical carcinoma cells is poorly understood. Therefore, we conducted a study to investigate whether anticancer drugs 5-FU and CP may exhibit the combined antiproliferative effect in cervical carcinoma cells. Using proteomics analysis, including two-dimensional gel electrophoresis and matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS), we investigated the antiproliferative effect-related proteins after treatment with 5-FU and/or CP. Our experiments showed that the combination of 5-FU and CP engaged both the apoptotic pathways: the membrane death receptor-mediated apoptosis pathway and the mitochondrial apoptotic pathway. Moreover, the combination of 5-FU and CP resulted in remarkable increasing susceptibility to apoptosis. We suggest that the combination of 5-FU and CP suppresses the growth of cervical carcinoma cells by synergistic effect with the induction of apoptosis. In vitro synergistic effect of 5-FU and CP supports the basis of the clinical application of the combination chemotherapy to the patients with cervical cancer.     

四物汤对放射线致血虚证小鼠骨髓蛋白质表达的影响

目的 :观察四物汤对血虚证小鼠骨髓蛋白质表达的影响 ,为阐明四物汤补血作用的分子机理提供理论和实验依据。方法 :采用 3.5Gy⁶⁰Coγ射线全身 1次性照射 ,制备小鼠血虚证模型。利用双向电泳、图像分析、胶内酶切、质谱鉴定等蛋白质组学技术 ,结合生物信息学 ,分离、分析、鉴定蛋白质。结果 :四物汤可以逆转放射线致血虚证小鼠骨髓 10个上调和 5个下调的蛋白质。其中 8个蛋白质可能分别是淋巴细胞特异性蛋白质 1、蛋白酶体26SATP酶亚组分 4、造血细胞蛋白质酪氨酸磷酸酶、H-ras、3-磷酸甘油醛脱氢酶、生长因子受体结合蛋白 14及Lgals12。结论 :四物汤能调节骨髓蛋白质表达 ,并可能由此促进造血细胞的生长和分化 ,发挥补血作用。     

Biomarkers in neonatology: the new “omics” of bronchopulmonary dysplasia

Bronchopulmonary dysplasia (BPD) is a complex disorder resulting from gene–environmental interactions. An improved understanding of the pathogenesis of this most common chronic lung disease in infants has been made by utilizing animal models and correlating with human data. Currently, while some (vitamin A, caffeine) pharmacotherapeutic options are being utilized to ameliorate this condition, there is still no specific or effective treatment for BPD. It would be helpful for prognostication and targeted potential novel therapeutic strategies to identify those babies accurately who are at risk for developing this disease. A reliable biomarker would have the capacity to be detected in the initial phase of the disease, to allow early interventions to avoid or minimize the detrimental effects of the disease. This review will focus on human studies performed with the “omic” techniques, specifically genomics, epigenomics, microbiomics, transciptomics, proteomics and metabolomics, and summarize the information available in the literature, as it pertains to biomarker identification for BPD. Using “omics” technologies, investigators have reported markers that have the potential to be used as biomarkers of BPD: SPOCK2, VEGF ?624C?>?G, VEGF ?460T?>?C, mast cells specific markers, miR-219 pathway, miR-152, ?30a-3p, ?133b, ?206, ?7, lactate, taurine, trimethylamine-N-oxide, gluconate, myoinositol and alterations in surfactant lipid profile.     

Hyaluronic acid‐dependent protection against UVB‐damaged human corneal cells

Within ultraviolet radiation, ultraviolet B (UVB) is the most energetic and damaging to humans. At the protein level, UVB irradiation downregulates the expression of antioxidant enzymes leading to the accumulation of reactive oxygen species (ROS). Due to lacking of a global analysis of UVB‐modulated corneal proteome, we investigate in vitro the mechanism of UVB‐induced corneal damage to determine whether hyaluronic acid (HA) is able to reduce UVB irradiation‐induced injury in human corneal epithelial cells. Accordingly, human corneal epithelial cell lines (HCE‐2) were irradiated with UVB, followed by incubation with low molecular weight HA (LMW‐HA, 100 kDa) or high molecular weight HA (HMW‐HA, 1,000 kDa) to investigate the physiologic protection of HMW‐HA in UVB‐induced corneal injury, and to perform a global proteomic analysis. The data demonstrated that HA treatment protects corneal epithelial cells in the UVB‐induced wound model, and that the molecular weight of HA is a crucial factor. Only HMW‐HA significantly reduces the UVB‐induced cytotoxic effects in corneal cells and increases cell migration and wound‐healing ability. In addition, proteomic analysis showed that HMW‐HA might modulate cytoskeleton regulation, signal transduction, biosynthesis, redox regulation, and protein folding to stimulate wound healing and to prevent these UVB‐damaged cells from cell death. Further studies evidenced membrane‐associated progesterone receptor component 1 (mPR) and malate dehydrogenase (MDH2) play essential roles in protecting corneal cells from UVB irradiation. This study reports on UVB‐modulated cellular proteins that might play an important role in UVB‐induced corneal cell injury and show HMW‐HA to be a potential substance for protecting corneal cells from UVB‐induced injury. Environ. Mol. Mutagen. 54:429–449, 2013. © 2013 Wiley Periodicals, Inc.     

Noticeable Decreased Expression of Tenascin-X in Calcific Aortic Valves

Calcification of aortic valves results in valvular aortic stenosis and is becoming a common valvular condition in elderly populations. An understanding of the molecular mechanisms of this valve lesion is important for revealing potential biomarkers associated with the development and progression of this disease. In order to identify proteins that are differentially expressed in calcific aortic valves (CAVs) compared with those in adjacent normal valvular tissues, comprehensive analysis of differentially expressed proteins in the tissues was done by a quantitative proteomic approach with isobaric tag for absolute and relative quantitation labeling followed by nanoliquid chromatography matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. The proteomic analysis revealed 105 proteins differentially expressed in CAVs in contrast to adjacent normal valvular tissues with high confidence. Significantly increased expression (>_1.3-fold) was found in 34 proteins, whereas decreased expression (<0.77-fold) was found in 39 proteins in CAVs. Among them, α-2-HS-glycoprotein showed the greatest increase in expression (6.54-fold) and tenascin-X showed the greatest decrease in expression (0.37-fold). Numerous extracellular matrix proteins such as collagens were identified as proteins with significantly decreased expression. Panther pathway analysis showed that some of the identified proteins were linked to blood coagulation and integrin signaling pathways. Cluster analysis of the 105 proteins differentially expressed in CAVs based on the expression pattern revealed that tenascin-X was clustered with proteins controlling collagen structure and function, especially collagen fibrillogenesis, such as decorin and fibromodulin. We confirmed decreased levels of these proteins in CAVs by Western blot analyses. These results indicated that massive destruction of the extracellular matrix occurs in CAVs.