Antitumor Activity of Choerospondias axillaris Fruit Extract by Regulating the Expression of SNCAIP and SNCA on MDA-MB-231 Cells

Objective: Cancer is a huge problem of disease globally. Today, the percentage of people die from cancer is more than a combination of various diseases. In females, most common types of malignancies that occur are breast and cervical. The present focus has been shifted on medicinal plants as a form of therapy and there is a constant need to identify new therapeutic agents. Choerospondias axillaris (C. axillaris), an underutilized fruit, has been used in the remedy of various diseases. In the present communication, we evaluated the molecular mechanism of C. axillaris methanol extract in regulating cell death in human breast cancer cells (MDA-MB-231). Methods: Methanol extract of C. axillaris was prepared and compounds were screened by Gas chromatography-mass spectrometry. The effect of fruit extract was determined on MDA-MB-231 cells by MTT ((3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) assay and to analyse the molecular mechanism of human breast cancer cells after treating with fruit extract, protein profiling study was performed by two-dimensional gel electrophoresis. Results: A total 9 differentially expressed proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS/MS) analysis. Among 9 identified proteins, synphilin-1 protein was found to be significantly downregulated, validated by western blot and RT-qPCR analysis. Possible interacting partners of synphilin-1 (SNCAIP) were analyzed for their possible role in cancer by the in-silico method. Conclusion: Our data implicate that the presence of bioactive compound(s) in C. axillaris fruits might play an important role in inhibiting the proliferation of breast carcinoma cells and Synphilin-1 protein may play a role of apoptotic function.     

Proteomic Profiling of Small Extracellular Vesicles Isolated from the Plasma of Vietnamese Patients with Non-Small Cell Lung Cancer Reveals Some Potential Biomarkers

Background: Considering the poor prognosis of non-small cell lung cancer (NSCLC), the objective of this study was to examine the potential of plasma-derived vesicles as a source of lung cancer-specific proteins. Extracellular vesicle (EV) cargos are specific to the source cells, hence they have the potential of being a source of cancer-specific proteins.  Methods: The proteins differently expressed in cancer were determined and derived from EVs isolated from the plasma of NSCLC patients at the National Lung Hospital. To this end, purification was done using gel filtration chromatography and ultracentrifugation. In addition, nano liquid chromatography mass spectrometry (LC–MS/MS) was used for analyzing. Results: Fifty-seven EV-derived proteins related to NSCLC were highlighted in this research. Some of them have not been addressed before, such as EEF1A1 (elongation factor 1-α1), KPNB1 (Importin subunit beta 1), SRC (proto-oncogene tyrosine-protein kinase) and ACTC1 (actin, alpha cardiac muscle 1). This list was further confirmed through a comparison with ExoCarta and Vesiclepedia. Conclusion: This study is the first work to show the involvement of several novel proteins of small EV (EEF1A1, KPNB1, SRC, and ACTC1) in the progression of NSCLC. The results suggested that they could serve as novel biomarkers for non-small cell lung cancer in the future.     

Pan-cancer proteomic map of 949 human cell lines

1. Download : Download high-res image (220KB)
2. Download : Download full-size image

     

Exosomes secreted by prostate cancer cells under hypoxia promote matrix metalloproteinases activity at pre-metastatic niches

Approximately, 30 000 men die from prostate cancer (PCa) every year in the United States, mainly due to the metastasis. Thus, the key events associated with PCa metastasis are under rigorous investigation, with recent studies showing that preparation of pre-metastatic niches (PMN) in distant organs is an important step. However, the molecular basis for PMN preparation is still unclear. Hypoxia in primary tumors promotes aggressiveness; however, its precise role in metastasis is not clear. We recently reported that exosomes secreted by PCa cells under hypoxia promote stemness and invasiveness in naïve PCa cells; however, whether these extracellular vesicles also influence PMN remains unknown. In the present study, we isolated exosomes from human PCa PC3 cells under normoxic (21% O₂, exosomes secreted under normoxic condition [Exo^Normoxic]) and hypoxic (1% O₂, exosomes secreted under hypoxic condition [Exo^Hypoxic]) conditions, and characterized their effect (10 µg exosomes, intraperitoneal (IP) treatment every 48 hours for 4 weeks) on key biomarkers associated with PMN in nude mice. Whole animal fluorescence imaging showed that Exo^Hypoxic treatment promotes matrix metalloproteinases (MMPs) activity in several putative metastatic sites. Histological studies confirmed that Exo^Hypoxic treatment enhanced the level of MMP2, MMP9, and extracellular matrix proteins (fibronectin and collagen) as well as increased the number of CD11b+ cells at selective PMN sites. Furthermore, proteomic profiling of exosomes by liquid chromatography/mass spectrometry identified cargo proteins in Exo^Normoxic and Exo^Hypoxic as well as distinct canonical pathways targeted by them. These results suggest that exosomes secreted by PCa cells under hypoxia plausibly remodel distant PMN, and thus, could be a potential target to control metastatic PCa.     

Two-dimensional gel electrophoresis (2D-GE) image analysis based on CellProfiler: Pseudomonas aeruginosa AG1 as model

Two-dimensional gel electrophoresis (2D-GE) is an indispensable technique for the study of proteomes of biological systems, providing an assessment of changes in protein abundance under various experimental conditions. However, due to the complexity of 2D-GE gels, there is no systematic, automatic, and reproducible protocol for image analysis and specific implementations are required for each context. In addition, practically all available solutions are commercial, which implies high cost and little flexibility to modulate the parameters of the algorithms. Using the bacterial strain, Pseudomonas aeruginosaAG1 as a model, we obtained images from 2D-GE of periplasmic protein profiles when the strain was exposed to multiple conditions, including antibiotics. Then, we proceeded to implement and evaluate an image analysis protocol with open-source software, CellProfiler. First, a preprocessing step included a bUnwarpJ-Image pipeline for aligning 2D-GE images. Then, using CellProfiler, we standardized two pipelines for spots identification. Total spots recognition was achieved using segmentation by intensity, whose performance was evaluated when compared with a reference protocol. In a second pipeline with the same program, differential identification of spots was addressed when comparing pairs of protein profiles. Due to the characteristics of the programs used, our workflow can automatically analyze a large number of images and it is parallelizable, which is an advantage with respect to other implementations. Finally, we compared six experimental conditions of bacterial strain in the presence or absence of antibiotics, determining protein profiles relationships by applying clustering algorithms PCA (Principal Components Analysis) and HC (Hierarchical Clustering).     

基于蛋白质组学分析色胺酮抑制小鼠乳腺癌的作用机制

目的:采用非标记定量(Label-free)蛋白质组学技术研究色胺酮抗小鼠体内乳腺癌的作用机制。方法:采用超高效液相色谱-质谱联用技术检测色胺酮抗小鼠乳腺癌的表达蛋白,选择Ionoptics nano UPLC C18色谱柱(0.075 mm×250 mm,1.6μm),流动相0.1%甲酸水溶液-0.1%甲酸乙腈溶液梯度洗脱,正离子模式,扫描范围m/z 100～1 700,使用MaxQuant 1.6.5.0进行数据库检索。采用Label-free高分辨质谱的蛋白质组学技术筛选4T1乳腺癌小鼠模型组与色胺酮(100 mg·kg~(-1))口服给药组之间的差异表达蛋白,进行色胺酮抗乳腺癌的蛋白质组学研究。结果:共鉴定出3 997个蛋白质,其中有2 911个蛋白可定量。模型组与色胺酮组共750个差异表达蛋白,其中286个蛋白上调,464个蛋白下调。基因本体分析表明,这些差异表达蛋白主要参与增殖、细胞迁移、凋亡、免疫、血管生成和炎症调节等生物学过程。京都基因与基因组百科全书通路分析进一步表明,这些蛋白主要集中于T细胞受体,B细胞受体,Toll样受体,核转录因子-κB(NF-κB),Ras蛋白,白细胞介素-17,肿瘤坏死因子,磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-Akt)和丝裂原活化蛋白激酶(MAPK)等信号通路。结论:与色胺酮抗4T1乳腺癌作用密切相关的差异表达蛋白包括上调蛋白白细胞分化抗原14(CD14),前列腺素G/H合酶2(PTGS2),泛素蛋白连接酶E3和下调蛋白CD44,70 kDa热休克蛋白1A(HSPA1A),巨噬细胞移动抑制因子(MIF),NF-κB,核糖体蛋白S6激酶α-4(RPS6KA4)和高迁移率族蛋白B1(HMGB1),提示色胺酮主要通过调节肿瘤炎症微环境来达到抑制小鼠乳腺癌的作用。     

Searching for prognostic biomarkers for small renal masses in the urinary proteome

Renal cell carcinoma (RCC) is frequently diagnosed incidentally as an early-stage small renal mass (SRM; pT1a, ≤4 cm). Overtreatment of patients with benign or clinically indolent SRMs is increasingly common and has resulted in a recent shift in treatment recommendations. There are currently no available biomarkers that can accurately predict clinical behavior. Therefore, we set out to identify early biomarkers of RCC progression. We employed a quantitative label-free liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) proteomics approach and targeted parallel-reaction monitoring to identify and validate early, noninvasive urinary biomarkers for RCC-SRMs. In total, we evaluated 115 urine samples, including 33 renal oncocytoma (≤4 cm) cases, 30 progressive and 26 nonprogressive clear cell RCC (ccRCC)-SRM cases, in addition to 26 healthy controls. We identified six proteins, which displayed significantly elevated expression in clear cell RCC-SRMs (ccRCC-SRMs) relative to healthy controls. Proteins C12ORF49 and EHD4 showed significantly elevated expression in ccRCC-SRMs compared to renal oncocytoma (≤4 cm). Additionally, proteins EPS8L2, CHMP2A, PDCD6IP, CNDP2 and CEACAM1 displayed significantly elevated expression in progressive relative to nonprogressive ccRCC-SRMs. A two-protein signature (EPS8L2 and CCT6A) showed significant discriminatory ability (areas under the curve: 0.81, 95% CI: 0.70–0.93) in distinguishing progressive from nonprogressive ccRCC-SRMs. Patients (Stage I–IV) with EPS8L2 and CCT6A mRNA alterations showed significantly shorter overall survival (p = 1.407 × 10⁻⁶) compared to patients with no alterations. Our in-depth proteomic analysis identified novel biomarkers for early-stage RCC-SRMs. Pretreatment characterization of urinary proteins may provide insight into early RCC progression and could potentially help assign patients to appropriate management strategies.     

Marine omega-3 fatty acid supplementation in non-alcoholic fatty liver disease: Plasma proteomics in the randomized WELCOME* trial

1. Download : Download high-res image (274KB)
2. Download : Download full-size image

     

基于蛋白质组学大黄异病同治急性中风大鼠脑物质的基础及相关机制

目的:基于定量蛋白质组学和生物信息学分析,探索大黄异病同治急性中风的物质基础及机制。方法:采用线栓法制备缺血再灌注的缺血性中风(IS)大鼠模型和胶原酶来诱导的出血性中风(ICH)模型。将60只SD大鼠随机分为缺血性中风假手术组(Sham1),缺血性中风组(IS),出血性中风+大黄治疗组(DH1),出血性中风假手术组(Sham2),出血性中风(ICH)组和出血性中风+大黄治疗组(DH2),每组10只。IS,Sham1和DH1组在24 h后,ICH,Sham2和DH2组在48 h后,生理盐水灌注后取脑组织行定量蛋白质组学分析,鉴定差异表达蛋白(DEPs)。对共性DEPs进行生物信息学分析,并对相关的DEPs进行蛋白免疫印迹验证。结果:大黄调节急性中风疾病相关共性DEPs 21个(上调12个,下调9个)。京都基因和基因组百科全书(KEGG)分析显示,富集了肌萎缩侧索硬化症(ALS)通路,通路中包含神经丝蛋白轻链多肽(Nefl),神经丝蛋白中链多肽(Nefm),神经丝蛋白重链多肽(Nefh)。大黄异病同治急性中风共性机制主要包括能量代谢、离子稳态、突触相关蛋白的调节、细胞周期及神经形成。共性DEPs验证,大黄治疗后,GTP结合蛋白REM2(Rem2),酪氨酸3-单加氧酶(Th),Nefl和神经调制蛋白(Gap43)表达量与相应模型组比较差异均有统计学意义(P0.05)。其中,治疗后Nefl表达为下调,而Rem2,Th和Gap43表达为上调,此结果与蛋白质组学检测结果一致。结论:该研究建立了大黄-异病同治-差异蛋白质表达谱,能量代谢、离子稳态、突触相关蛋白调节、细胞周期及神经形成是其共性机制。