Comparison of RNAscope and immunohistochemistry for evaluation of the UPK2 status in urothelial carcinoma tissues

Abstract

Background UPK2 exhibits excellent specificity for urothelial carcinoma (UC). UPK2 evaluation can be useful in making the correct diagnosis of UC. However, UPK2 detection by immunohistochemistry (IHC) has relatively low sensitivity. This paper aimed to compare the diagnostic sensitivity of RNAscope and IHC for evaluation of the UPK2 status in UC. Methods Tissue blocks from 127 conventional bladder UCs, 45 variant bladder UCs, 24 upper tract UCs and 23 metastatic UCs were selected for this study. IHC and RNAscope were used to detect the UPK2 status in UCs. Then, comparisons of the two methods were undertaken. Results There was no significant difference between RNAscope and IHC for the evaluation of the UPK2 positivity rate in UC (68.0% vs. 62.6%, P = 0.141). Correlation analysis revealed a moderate positive correlation for detection of UPK2: RNAscope vs. IHC (P < 0.001, R = 0.441). Our results showed a trend toward a higher positive UPK2 rate detected by RNAscope (53.3%) than by IHC (35.6%) in variant bladder UCs. Disappointingly, the P value did not indicate a significant difference (P = 0.057). Conclusions RNAscope for UPK2 appeared to perform similarly to IHC, with a marginally higher positive rate, suggesting it could be used as an alternative or adjunct to UPK2 IHC.

A Multi-Tissue Comparison and Molecular Characterization of Canine Organoids

“…Desmoglein 3 (DSG3, FWER = 2.28 × 10 -3 ), which is a basal cell marker (Elbadawy et al, 2019), and Loricrin cornified envelope precursor protein (LORICRIN, FWER = 1.38 × 10 -2 ), an intermediate cell marker (Lin et al, 2013), were upregulated in organoids compared to tissues ( Figure 2A ). Uroplakin (UPK) proteins are specific to urothelial cells (Lu et al, 2022), and both Uroplakin 2 (UPK2, FWER = 1.70 × 10 -2 ) and Uroplakin 3A (UPK3A, FWER = 4.74 × 10 -3 ) were upregulated in organoids ( Figure 2B ). UPK3A is an umbrella cell marker, while Keratin 5 (KRT5) is indicative of basal cells and was bladder-specific and upregulated in the canine bladder organoids (Elbadawy et al, 2022; Kim et al, 2020a).…”

Recent Blood and Intracranial Molecular Biomarkers to Moderate Depression and Major Depressive Disorder

“…With the development of IHC technology, it plays an increasingly important role in clinical medicine, especially in the detection and treatment of cancer [35]. In the research of thyroid cancer, IHC technology plays an important clinical application [36]. Papillary thyroid cancer (PTC) is a common thyroid malignancy.…”

“…In molecular diagnosis based on RNA ISH, RNA is easily degraded, which greatly limits the development of RNA ISH technology [36]. The RNAscope technology, as an improvement of the RNA in situ hybridization (ISH) method, is a quantitative in situ gene expression measurement technology [38].…”

“…However, compared with the newly discovered RNAscope technology, IHC has shown great limitations. The biggest limitation is that the accuracy of IHC is affected by the density of targeted cells and tissues and the sensitivity limits of the detection system [36]. Taking the detection of breast cancer as an example, when immunohistochemistry (IHC) is used in clinical experiments to determine the disease state of patients, it is found that this method is highly subjective, requires a large amount of tumor tissue, and is prone to false negative results.…”

“…The current laboratory application further expands the application of RNAscope detection technology in pathological diagnosis and differential diagnosis. Especially for a set of highly specific, diagnostic IHC markers, RNAscope can detect some specific, organ-specific biomarkers, and may also serve as another example in the event that IHC fails to show a detectable signal [36]. Therefore, considering the heterogeneity of individual tissues, IHC can be used in combination with RNAscope technology.…”