scholarly journals An integrated transcriptomics and proteomics study of Head and Neck Squamous Cell Carcinoma – methodological and analytical considerations.

2015 ◽  
Author(s):  
Anupama Rajan Bhat ◽  
Manoj Kumar Gupta ◽  
Priya Krithivasan ◽  
Kunal Dhas ◽  
Jayalakshmi Nair ◽  
...  

High throughput molecular profiling and integrated data analysis with tumor tissues require overcoming challenges like tumor heterogeneity and tissue paucity. This study is an attempt to understand and optimize various steps during tissue processing and in establishing pipelines essential for integrated analysis. Towards this effort, we subjected laryngo-pharyngeal primary tumors and the corresponding adjacent normal tissues (n=2) to two RNA and protein isolation methods, one wherein RNA and protein were isolated from the same tissue sequentially (Method 1) and second, wherein the extraction was carried out using two independent methods (Method 2). RNA and protein from both methods were subjected to RNA-seq and iTRAQ based LC-MS/MS analysis. Transcript and peptide identification and quantification was followed by both individual -ome and integrated data analysis. As a result of this analysis, we identified a higher number of total, as well as differentially expressed (DE) transcripts (1329 vs 1134) and proteins (799 vs 408) with fold change ≥ 2.0, in Method 1. Among these, 173 and 86 entities were identified by both transcriptome and proteome analysis in Method 1 and 2, respectively, with higher concordance in the regulation trends observed in the former. The significant cancer related pathways enriched with the individual DE transcript or protein data were similar in both the methods. However, the entities mapping to them were different, allowing enhanced view of the pathways identified after integration of the data and subsequent mapping. The concordant DE transcripts and proteins also revealed key molecules of the pathways with important roles in cancer development. This study thus demonstrates that sequential extraction of the RNA and proteins from the same tissue allows for better profiling of differentially expressed entities and a more accurate integrated data analysis.

2019 ◽  
Vol 16 (2) ◽  
pp. 148-155
Author(s):  
Asma Tariq ◽  
Rana Muhammad Mateen ◽  
Iram Fatima ◽  
Muhammad Waheed Akhtar

Objective: The aim of the present study was to build protein profiles of untreated breast cancer patients of invasive ductal carcinoma grade II at tissue level in Pakistani population and to compare 2-D profiles of breast tumor tissues with matched normal tissues in order to evaluate for variations of proteins among them. Materials & Methods: Breast tissue profiles were made after polytron tissue lysis and rehydrated proteins were further characterized by using two-dimensional gel electrophoresis. On the basis of isoelectric point (pI) and molecular weight, proteins were identified by online tool named Siena 2-D database and their identification was further confirmed by using MALDI-TOF. Results: Among identified spots, 10 proteins were found to be differentially expressed i.e.; COX5A, THIO, TCTP, HPT, SODC, PPIA, calreticulin (CRT), HBB, albumin and serotransferrin. For further investigation, CRT was selected. The level of CRT in tumors was found to be significantly higher than in normal group (p < 0.05). The increased expression of CRT level in tumor was statistically significant (p = 0.010) at a 95% confidence level (p < 0.05) as analyzed by Mann-Whitney. CRT was found distinctly expressed in high amount in tumor tissue as compared to their matched normal tissues. Conclusion: It has been concluded that CRT expression could discriminate between normal tissue and tumor tissue so it might serve as a possible candidate for future studies in cancer diagnostic markers.


2020 ◽  
Author(s):  
Zixi Chen ◽  
Jinfen Wei ◽  
Yuchen Yuan ◽  
Ying Cui ◽  
Yanyu Zhang ◽  
...  

AbstractBackgroundMetabolism reprogramming and immune evasion are the most fundamental hallmarks for cancer survival. The complex interactions between metabolism and immune systems in tumors and their microenvironment is complicated. Researching on the correlation changes between metabolic and immune related-genes in normal and tumor tissues would help to reveal these complex interactions.MethodsIn this study, the mRNA profiles across 11 cancer types was obtained from The Cancer Genome Atlas (TCGA). Then, the spearman’s correlation coefficient was calculated between metabolic and immune related-genes for each sample group.ResultsOur results showed that the number of correlated gene pairs was reduced significantly in tumor tissues compared with those of normal tissue, especially in KIRC, KIRP and STAD. Functional enrichment analysis for the universal (the pairs appeared in more than 2 cancer types) and specific (the pairs only in one specific cancer type) gene pairs across cancer types revealed top pathways which appeared in tumor and normal samples, such as phosphatidylinositol signaling system and inositol phosphate metabolism. Thereinto, the pairs in normal tissues missing in tumors may indicate they are important factors affecting immune system, such as, DGKs and PIP4ks. The correlation analysis between immune checkpoint and metabolism genes also showed a reduced correlation in tumor and had the tissue specificity, such as, FUT8 was strongly correlated with PDCD1 in the HC of STAD and they had a weaker correlation in other normal tissues and tumor types.ConclusionsOur study provides a novel strategy for investigating interaction of tumor immune and metabolism in microenvironment and offers some key points for exploring new targets including metabolic targets and immunomodulator of immune checkpoints.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Meini Wu ◽  
Wenliang Li ◽  
Fengchang Huang ◽  
Jing Sun ◽  
Kang ping Li ◽  
...  

AbstractLong non-coding RNAs (lncRNAs) act as competing endogenous RNAs (ceRNAs) to compete with microRNAs (miRNAs) in cancer occurrence and development. However, the differential expression of RNAs and their ceRNA network during the development of colon cancer (CC) remains unclear. This study was aimed at comprehensive analysis of the lncRNAs and their ceRNA networks associated with CC. Whole transcriptome sequencing was performed on colorectal and adjacent normal tissues at different pathological stages. Forty-nine lncRNAs were differently expressed between the CC tissues and their adjacent normal tissues at all stages. Aberrant expression of lncRNA CDKN2B-AS1 and lncRNA MIR4435-2HG was confirmed by TCGA database. Moreover, 14 lncRNAs were differentially expressed between early and advance stages of the tumor tissues, and 117 miRNAs were specifically expressed in stage III & IV. Weighted gene co-expression network analysis of 17105 differently expressed mRNAs revealed that the mRNAs shown in module pink, midnight blue, black, and light cyan were related to TNM and pathological stage, and that these mRNAs were enriched in cancer related functions and pathways. As DElncRNA showed a trend of change similar to that of the DEmRNA and opposite to that of DEmiRNA, ceRNA network was constructed with 3 DEmiRNAs, 5 DElncRNAs, and 130 DEmRNAs. Real time PCR revealed that expression of MEG3 was decreased in the tumor tissues belonging to stage III and IV as compared to that in stage I. Moreover, hsa-miR-324-5p was upregulated, while FGFR3, PLCB4, and IKBKB were downregulated in the tumor tissues as compared to that in the adjacent normal tissues. Thus, this study revealed differentially expressed lncRNA between different stages of CC as well as suggested that lncRNA CDKN2B-AS1, MIR4435-2HG, and MEG3 may act as diagnostic biomarkers for the development of CC.


2020 ◽  
Author(s):  
Lin Chen ◽  
Heng Sun ◽  
Jie Kong ◽  
Haijiang Xu ◽  
Xiyan Yang

Abstract BackgroundSoil salt stress seriously restricts the yield and quality of cotton worldwide. To investigate the molecular mechanism of cotton response to salt stress, a main cultivated variety Gossypium hirsutum L. acc. Xinluzhong 54 was used to perform transcriptome and proteome integrated analysis. ResultsThrough transcriptome analysis of cotton treated with salt stress for 0 h (T0), 3 h (T3) and 12 h (T12), we identified 8,436, 11,628 and 6,311 differentially expressed genes (DEGs) inT3 / T0, T12 / T0 and T12 / T3, respectively. A total of 459 differentially expressed proteins (DEPs) were identified by proteomic analysis, of which 273, 99 and 260 DEPs were identified in T3 / T0, T12 / T0 and T12 / T3, respectively. Metabolic pathways, biosynthesis of secondary metabolites, photosynthesis and plant hormone signal transduction were the main enrichment pathways by annotation of DEGs or DEPs. Detail analysis of the DEGs or DEPs revealed that complex signal pathways, such as ABA and JA signal, calcium signal, MAPK signal cascade, transcription factors, followed by activation of antioxidant and ion transporters, were identified to participate in regulating salt response in cotton.ConclusionsOur results not only contribute to understand the mechanism of cotton response to salt stress, but also provide nine candidate genes, which might be used for molecular breeding to improve salt-tolerance in cotton.


2021 ◽  
Author(s):  
Yili Ren ◽  
Beibei Zhang ◽  
Chenkai Xu ◽  
Lei Zhang

Abstract Background and purpose: Gastric cancer is a type of highly heterogeneous malignant tumor and the prognosis of gastric cancer is hard to be improved due to limited knowledge on the molecular mechanism of heterogeneity. Single-cell sequencing technology is recently widely used for the investigation of both inter-tumoral heterogeneity and intra-tumoral heterogeneity. The present study aims to explore the potential oncogene by analyzing the single-cell data in the GSE167297 dataset.Methods: The GSE167297 dataset was downloaded from the GEO database, followed by quality control to remove data with lower quality. The division on cell subtypes was determined by the characteristic marker expressed in each cell subpopulation. Wilcoxon rank-sum test was used to screen out differentially expressed genes. Survival analysis was performed to evaluate the prognostic value of G-protein subunit g 11 (GNG11) gene which was significantly overexpressed in deep tumor tissues of diffuse gastric cancer.Results: In both normal tissues and tumor tissues, subtypes of immune cells and stromal cells were identified, with a higher proportion of infiltrated macrophages observed in deep tumor tissues. EPCAM was found significantly highly expressed in a cell subpopulation from gastric tumor tissues. 515 differentially expressed genes (| log2FC | > 2 and FDR < 1e-5) were screened out between normal tissues and tumor tissues. 86 differentially expressed genes (| log2FC | > 1 and FDR < 0.01) were screened out between superficial and deep tumor tissues, in which GNG11 was most highly expressed in deep tumor tissues (mean expression value: 0.1247, FC value: 52.2109). Disease-specific survival analysis on GNG11 results showed that the HR [95%CI] in the constructed univariate Cox proportional risk model was 4.419 [1.399-13.96] and the P-value in the log-rank test was 0.0056.Conclusion: Differentially expression profiles were provided both extratumorally and intratumorally, indicating a higher infiltration of macrophages in deep tumor tissues. Additionally,GNG11 was screened out to be a significant risk factor in STAD patients.


2020 ◽  
Author(s):  
Xinghao Chen ◽  
Hanqi Liu ◽  
Shijie Wang ◽  
Chao Zhang ◽  
Minsheng Yang ◽  
...  

Abstract BackgroundThe red-leaved poplar cultivars ‘Quanhong’ and ‘Xuanhong’ are bud mutations of Populus deltoides cv. ‘Zhonglin 2025’. These cultivars are valued for their beautiful shape, lack of flying catkins, and ornamental leaf colors. The molecular mechanism that leads to different leaf colors between these red-leaved poplars and ‘Zhonglin 2025’ remains unclear.ResultsIn this study, we analyzed growth and physiological indicators in the ‘Quanhong’, ‘Xuanhong’, and ‘Zhonglin 2025’ poplar cultivars, and performed transcriptome and proteome analysis of their leaves. The results showed that plant height and ground diameter were significantly lower in both red-leaved poplars than in ‘Zhonglin 2025’, indicating that their growth and development were markedly inhibited. The ratio of anthocyanin to total chlorophyll and photosynthetic capacity of the leaves were higher and lower, respectively, in the red-leaved cultivars than in ‘Zhonglin 2025’. At the transcript and protein levels, 6,792 differentially expressed genes (DEGs) and 2,786 differentially expressed proteins (DEPs) were screened in the ‘Quanhong’ cultivar, respectively, and 4,398 DEGs and 2,333 DEPs were screened in the ‘Xuanhong’ cultivar, respectively. We screened 769 DEGs/DEPs in ‘Quanhong’ and 399 DEGs/DEPs in ‘Xuanhong’ in an integrated transcriptomics/proteomics analysis. Based on the results of this integrated analysis, 15 and 11 genes/proteins involved in anthocyanin synthesis were further identified in ‘Quanhong’ and ‘Xuanhong’, respectively, including the CHS , F3H , and DFR genes. Among the 120 transcription factors, 3 (HY5, HYH, and TTG2), may be directly involved in the regulation of anthocyanin synthesis in both red-leaved poplars.ConclusionsBy comparing the proteomes and transcriptomes of red-leaved poplar cultivars and ‘Zhonglin 2025’, we identified the key genes and proteins related to red leaf coloration. The findings of this study provided insights that may aid further studies of the molecular mechanism of leaf red coloration in red-leaved poplars.


2021 ◽  
Vol 2021 ◽  
pp. 1-14
Author(s):  
Yu Hu ◽  
Hongyan Zai ◽  
Wei Jiang ◽  
Yuanbing Yao ◽  
Zhenglin Ou ◽  
...  

Background. Extracellular vesicles (EVs) can deliver miRNAs between cells and play a crucial role in hepatoblastoma progression. In this study, we explored the differentially expressed miRNAs related to tumor cell-derived EVs and the mechanism by which EVs regulate hepatoblastoma progression. Methods. Bioinformatics analysis was performed to explore the differentially expressed miRNAs between the hepatoblastoma and adjacent normal tissues. TEM, NTA, and western blotting were conducted to identify EVs. The expression of miR-126-3p, miR-126-5p, miR-30b-3p, miR-30b-3p, SRY, IL-1α, IL-6, and TGF-β was detected by RT-qPCR. Immunofluorescence (IF) was used to analyze the expression of PKH67, and flow cytometry was applied to assess the ratio of CD44+ CD90+ CD133+ cells. ELISA was used to evaluate the levels of IL-6 and TGF-β. A xenograft mouse model was constructed to detect the function of EVs with downregulated miR-126. IHC was performed to calculate β-catenin levels in tumor tissues. Results. miR-126 was upregulated in hepatoblastoma. EVs derived from hepatoblastoma cells significantly increased the ratio of CD44+ CD90+ CD133+ cells and increased the expression of IL-6, Oct4, SRY, and TGF-β in bone marrow mesenchymal stem cells (BMSCs), while EVs with downregulated miR-126 reversed these phenomena. miR-126 downregulation notably attenuated hepatoblastoma tumor growth and decreased the ratio of CD44+ CD90+ CD133+ cells and increased the expression of IL-6, Oct4, SRY, TGF-β, and β-catenin in tumor tissues of mice. Furthermore, EVs with downregulated miR-126 inhibited the differentiation of BMSCs into cancer stem cells. Conclusions. Exosomal miR-126 derived from hepatoblastoma cells promoted the tumorigenesis of liver cancer through inducing the differentiation of BMSCs into cancer stem cells.


2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Qiancheng Qiu ◽  
Yazhen Li ◽  
Zhiqiang Fan ◽  
Fen Yao ◽  
Wenjun Shen ◽  
...  

Purpose. Human papillomavirus (HPV) antigens had been found in colorectal cancer (CRC) tissue, but little evidence demonstrates the association of HPV with oncogene mutations in CRC. We aim to elucidate the mutated genes that link HPV infection and CRC carcinogenesis. Methods. Cancerous and adjacent noncancerous tissues were obtained from CRC patients. HPV antigen was measured by using the immunohistochemical (IHC) technique. The differentially expressed genes (DEGs) in HPV-positive and HPV-negative tumor tissues were measured by using TaqMan Array Plates. The target genes were validated with the qPCR method. Results. 15 (31.9%) cases of CRC patients were observed to be HPV positive, in which HPV antigen was expressed in most tumor tissues rather than in adjacent noncancerous tissues. With TaqMan Array Plates analyses, we found that 39 differentially expressed genes (DEGs) were upregulated, while 17 DEGs were downregulated in HPV-positive CRC tissues compared with HPV-negative tissues. Four DEGs (MMP-7, MYC, WNT-5A, and AXIN2) were upregulated in tumor vs. normal tissues, or adenoma vs. normal tissue in TCGA, which was overlapped with our data. In the confirmation test, MMP-7, MYC, WNT-5A, and AXIN2 were upregulated in cancerous tissue compared with adjacent noncancerous tissue. MYC, WNT-5A, and AXIN2 were shown to be upregulated in HPV-positive CRC tissues when compared to HPV-negative tissues. Conclusion. HPV-encoding genome may integrate into the tumor genomes that involved in multiple signaling pathways. Further genomic and proteomic investigation is necessary for obtaining a more comprehensive knowledge of signaling pathways associated with the CRC carcinogenesis.


2020 ◽  
pp. mcp.RA120.002384
Author(s):  
Juntuo Zhou ◽  
Bing Liu ◽  
Zhongwu Li ◽  
Yang Li ◽  
Xi Chen ◽  
...  

The histopathological subtype of lung adenocarcinoma (LUAD) is closely associated with prognosis. Micropapillary or solid predominant LUAD tends to relapse after surgery at an early stage, whereas lepidic pattern shows a favorable outcome. However, the molecular mechanism underlying this phenomenon remains unknown. Here, we recruited 31 lepidic predominant LUADs (LR: low-risk subtype group) and 28 micropapillary or solid predominant LUADs (HR: high-risk subtype group). Tissues of these cases were obtained and label-free quantitative proteomic and bioinformatic analyses were performed. Additionally, prognostic impact of targeted proteins was validated using The Cancer Genome Atlas databases (n=492) and tissue microarrays composed of early-stage LUADs (n=228). A total of 192 differentially expressed proteins were identified between tumor tissues of LR and HR and three clusters were identified via hierarchical clustering excluding eight proteins. Cluster 1 (65 proteins) showed a sequential decrease in expression from normal tissues to tumor tissues of LR and then to HR and was predominantly enriched in pathways such as tyrosine metabolism and ECM-receptor interaction, and increased matched mRNA expression of 18 proteins from this cluster predicted favorable prognosis. Cluster 2 (70 proteins) demonstrated a sequential increase in expression from normal tissues to tumor tissues of LR and then to HR and was mainly enriched in pathways such as extracellular organization, DNA replication and cell cycle, and high matched mRNA expression of 25 proteins indicated poor prognosis. Cluster 3 (49 proteins) showed high expression only in LR, with high matched mRNA expression of 20 proteins in this cluster indicating favorable prognosis. Furthermore, high expression of ERO1A and FEN1 at protein level predicted poor prognosis in early-stage LUAD, supporting the mRNA results. In conclusion, we discovered key differentially expressed proteins and pathways between low-risk and high-risk subtypes of early-stage LUAD. Some of these proteins could serve as potential biomarkers in prognostic evaluation.


2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Lin CHEN ◽  
Heng SUN ◽  
Jie KONG ◽  
Haijiang XU ◽  
Xiyan YANG

Abstract Background Soil salt stress seriously restricts the yield and quality of cotton worldwide. To investigate the molecular mechanism of cotton response to salt stress, a main cultivated variety Gossypium hirsutum L. acc. Xinluzhong 54 was used to perform transcriptome and proteome integrated analysis. Results Through transcriptome analysis in cotton leaves under salt stress for 0 h (T0), 3 h (T3) and 12 h (T12), we identified 8 436, 11 628 and 6 311 differentially expressed genes (DEGs) in T3 vs. T0, T12 vs. T0 and T12 vs. T3, respectively. A total of 459 differentially expressed proteins (DEPs) were identified by proteomic analysis, of which 273, 99 and 260 DEPs were identified in T3 vs. T0, T12 vs. T0 and T12 vs. T3, respectively. Metabolic pathways, biosynthesis of secondary metabolites, photosynthesis and plant hormone signal transduction were enriched among the identified DEGs or DEPs. Detail analysis of the DEGs or DEPs revealed that complex signaling pathways, such as abscisic acid (ABA) and jasmonic acid (JA) signaling, calcium signaling, mitogen-activated protein kinase (MAPK) signaling cascade, transcription factors, activation of antioxidant and ion transporters, were participated in regulating salt response in cotton. Conclusions Our research not only contributed to understand the mechanism of cotton response to salt stress, but also identified nine candidate genes, which might be useful for molecular breeding to improve salt-tolerance in cotton.


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