Methods in molecular biology
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Technological advances in the form of next-generation sequencing allow sequencing of large numbers of different DNA sequences in a single/parallel reaction compared to conventional sequencing. It is a powerful tool which has enabled comprehensive characterization of esophageal squamous cell carcinoma. Whole-genome sequencing is the most comprehensive but expensive, whereas whole-exome sequencing is cost-effective, but it only works for the known genes. Thus, second-generation sequencing methods can provide a complete picture of the esophageal squamous cell carcinoma genome by detecting and discovering different type of alterations in the cancer which may lead to the development of effective diagnostic and therapeutic approaches for esophageal squamous cell carcinoma.
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Tissue microarray (TMA) is widely used for identifying the expression of markers in many tissues from patients with esophageal squamous cell carcinoma. The technology is mostly used in immunohistochemical studies to test the expression of markers and oncoproteins in signalling pathway as well as targeting proteins involved in therapies for esophageal squamous cell carcinoma. Appropriate use of TMA sections needs consideration of labor, planning, and expertise involved. For the best performance, it is important to design the layout of the TMA as well as use whole-slide scanning for interpretation of the TMA sections.
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Macroscopic examination of the surgical specimen of esophageal squamous cell carcinoma by pathologist is important for quality clinical management, research, as well as education purposes. The process includes dissection of the specimen, identification of the lesion, measurements, and taking appropriate samples for histopathological examination. ⋯ In addition, examination of proximal resection margin and circumferential resection margin are important in the cancer. A standardized approach for macroscopic examination by professionals is needed for accurate diagnosis and to optimize the use of the surgical specimen with esophageal squamous cell carcinoma.
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Whole-slide imaging (WSI) contributes to medical education, collaboration, quality assurance, examination, and consultation in pathology. The images obtained from WSI are of high quality and could be stored indefinitely. ⋯ The operation and basic principles of whole-slide imaging of esophageal squamous cell carcinoma are also presented. Common use of WSI will occur with modifications of the whole-slide imaging scanners to adapt to the workflows in diagnostic and research laboratories.
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MicroRNAs (miRNAs) are 20-22 nucleotides long single-stranded noncoding RNAs. They regulate gene expression posttranscriptionally by base pairing with the complementary sequences in the 3'-untranslated region of their targeted mRNA. Aberrant expression of miRNAs leads to alterations in the expression of oncogenes and tumor suppressors, thereby affecting cellular growth, proliferation, apoptosis, motility, and invasion capacity of gastrointestinal cells, including cells of esophageal squamous cell carcinoma (ESCC). ⋯ Consequently, expression profiles of miRNAs could be useful as diagnostic, prognostic, and prediction biomarkers in ESCC. Herein, we describe the quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and microarray methods for detection and quantitate miRNAs in ESCC. In addition, we summarize the roles of miRNAs in ESCC pathogenesis, progression, and prognosis.