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The expression level of microRNA (miR)-92a has been proven to increase during the development of colorectal adenocarcinoma (CA) and has been verified at the cellular, plasma and fecal levels by various quantitative methods. However, a method to quantitate the expression level using tissue sections has not been established. To do this, in situ hybridization (ISH) and multispectral imaging microscopy (MSI) were introduced to quantitate miR-92a expression on the microscopic level. ISH of miR-92a was first performed on 34 tissue samples of CA and adenomas with high-grade and low-grade intraepithelial neoplasms, while 31 paralesional normal tissue samples were defined as the control. Subsequently, a MSI technique was applied to quantitate the hybridization signal in terms of optical density (OD) at the visible wavelength. A t-test with unequal variance was used to examine the statistical significance between the groups. Despite all 34 tissue sections demonstrating at least partial positivity of miR-92a expression following ISH, visual grading was inconclusive. As such, the signal of ISH was transformed in terms of OD and further analyzed by employing the MSI system. A statistically significant difference was observed between the expression levels of miR-92a in CA and the paralesional normal controls. By contrast, a poor correlation was revealed between visual and spectral grading. The co-utilization of ISH and MSI generated a legible observation in the expression level of miR-92a, revealing the dynamic change in miR-92a expression in the progression of the disease and providing important information for further functional investigation.

Quantitation of microRNA-92a in colorectal adenocarcinoma and its precancerous lesions: Co-utilization of in situ hybridization and spectral imaging