scholarly journals Analysis of mutations in tumor and normal adjacent tissue via fluorescence detection

Author(s):  
Jennifer E. Kay ◽  
Sheyla Mirabal ◽  
William E. Briley ◽  
Takafumi Kimoto ◽  
Theofilos Poutahidis ◽  
...  
2016 ◽  
Author(s):  
Patricia Bortman Rozenchan ◽  
Cristina V. de Carvalho ◽  
Tatiana Bonetti ◽  
Carina Melo ◽  
M Mitzi Brentani ◽  
...  

2012 ◽  
Vol 30 (5_suppl) ◽  
pp. 315-315
Author(s):  
Wei Meng ◽  
Arnab Chakravarti ◽  
Tim Lautenschlaeger

315 Background: DNA methylation and histone modification are widely studied epigenetic events that can decrease gene expression levels. Promoter hypermethylation has been proposed as a potential diagnostic or prognostic biomarker in various cancers. We studied DNA promoter methylation of 5 cancer-associated genes (MRE11, APX1, ERCC1, RASSF1A and RASSF2A ) in paired FFPE bladder cancer tissues and normal adjacent tissue. The MRE11/Rad50/NBS1 complex serves as a single-strand DNA nuclease which participates in the repair of DNA double-strand breaks and replication errors. APX1 plays a key role in regulating H2O2 levels and H2O2 signaling. DNA repair machinery, ERCC1 protein levels in tumor tissue have been shown to predict response to platinum-based chemotherapy. RASSF1 is thought to be a tumor suppressor gene, while the function of RASSF2 is less well understood. Methods: 16 bladder cancer cases with available paraffin-embedded tumor and matched normal adjacent tissue specimens (32 tissue samples) were analyzed. DNA was extracted by Ambion RecoverAll Total Nucleic Acid Isolation kit. FFPE DNA bisulfite modification was performed using Zymo EZ DNA Methylation Kit. Methylation Specific-High Resolution Melting (MS-HRM) analysis was used to assess methylation status. MS-HRM monitors the melting behavior of PCR amplicons by using a DNA intercalating fluorescent dye. Results: MRE11 and APX1 promoters were not methylated in either cancer or normal adjacent samples. The ERCC1 gene was heavily methylated in 94% (30/32) of all cancer and wild type samples. RASSF1A and RASSF2A promoter methylation was significantly different between cancer and normal adjacent tissue. 50% (8/16) RASSF1A and 25% (4/16) RASSF2A had promoter methylation in cancer tissues, while only 6% (1/16) RASSF1A and 0% (0/16) RASSF2A had promoter methylation in normal adjacent tissues. 69% (11/16) of bladder cancer tissues were positive for RASSF1A or RASSF2A promotor methylation while only 1/16 normal adjacent tissue samples was positive for either promotor methylation. Conclusions: The results show that cancer and non-cancer tissue have different RASSF1A and RASSF2A promoter methylation patterns.


2002 ◽  
Vol 26 (3) ◽  
pp. 238-244 ◽  
Author(s):  
Christopher J Sweeney ◽  
Mark S Marshall ◽  
Darlene S Barnard ◽  
Douglas K Heilman ◽  
Steven D Billings ◽  
...  

Cancer ◽  
1981 ◽  
Vol 47 (12) ◽  
pp. 2883-2886 ◽  
Author(s):  
Herman Baker ◽  
Oscar Frank ◽  
Thomas Chen ◽  
Susan Feingold ◽  
Barbara Deangelis ◽  
...  

Oncogene ◽  
2009 ◽  
Vol 28 (49) ◽  
pp. 4375-4385 ◽  
Author(s):  
C Peña ◽  
J M García ◽  
M J Larriba ◽  
R Barderas ◽  
I Gómez ◽  
...  

2020 ◽  
Vol 2020 ◽  
pp. 1-6 ◽  
Author(s):  
Michael A. Liss ◽  
Yidong Chen ◽  
Ronald Rodriguez ◽  
Deepak Pruthi ◽  
Teresa Johnson-Pais ◽  
...  

Objective. To perform a proof of concept microbiome evaluation and PD-L1 expression profiling in clear-cell renal cell carcinoma (cc-RCC) with associated tumor thrombus (TT). Methods. After IRB approval, six patients underwent radical nephrectomy (RN) with venous tumor thrombectomy (VTT). We collected fresh tissue specimens from normal adjacent, tumor, and thrombus tissues. We utilized RNA sequencing to obtain PD-L1 expression profiles and perform microbiome analysis. Statistical assessment was performed using Student’s t-test, chi-square, and spearman rank correlations using SPSS v25. Results. We noted the tumor thrombus to be mostly devoid of diverse microbiota. A large proportion of Staphylococcus epidermidus was detected and unknown if this is a surgical or postsurgical contaminant; however, it was noted more in the thrombus than other tissues. Microbiome diversity profiles were most abundant in the primary tumor compared to the thrombus or normal adjacent tissue. Differential expression of PD-L1 was examined in the tumor thrombus to the normal background tissue and noted three of the six subjects had a threshold above 2-fold. These three similar subjects had foreign microbiota that are typical residents of the oral microbiome. Conclusion. Renal tumors have more diverse microbiomes than normal adjacent tissue. Identification of resident oral microbiome profiles in clear-cell renal cancer with tumor thrombus provides a potential biomarker for thrombus response to PD-L1 inhibition.


2016 ◽  
Vol 82 (16) ◽  
pp. 5039-5048 ◽  
Author(s):  
Camilla Urbaniak ◽  
Gregory B. Gloor ◽  
Muriel Brackstone ◽  
Leslie Scott ◽  
Mark Tangney ◽  
...  

ABSTRACTIn the United States, 1 in 8 women will be diagnosed with breast cancer in her lifetime. Along with genetics, the environment contributes to disease development, but what these exact environmental factors are remains unknown. We have previously shown that breast tissue is not sterile but contains a diverse population of bacteria. We thus believe that the host's local microbiome could be modulating the risk of breast cancer development. Using 16S rRNA amplicon sequencing, we show that bacterial profiles differ between normal adjacent tissue from women with breast cancer and tissue from healthy controls. Women with breast cancer had higher relative abundances ofBacillus,EnterobacteriaceaeandStaphylococcus.Escherichia coli(a member of theEnterobacteriaceaefamily) andStaphylococcus epidermidis, isolated from breast cancer patients, were shown to induce DNA double-stranded breaks in HeLa cells using the histone-2AX (H2AX) phosphorylation (γ-H2AX) assay. We also found that microbial profiles are similar between normal adjacent tissue and tissue sampled directly from the tumor. This study raises important questions as to what role the breast microbiome plays in disease development or progression and how we can manipulate this for possible therapeutics or prevention.IMPORTANCEThis study shows that different bacterial profiles in breast tissue exist between healthy women and those with breast cancer. Higher relative abundances of bacteria that had the ability to cause DNA damagein vitrowere detected in breast cancer patients, as was a decrease in some lactic acid bacteria, known for their beneficial health effects, including anticarcinogenic properties. This study raises important questions as to the role of the mammary microbiome in modulating the risk of breast cancer development.


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