| Objective: Estrogen receptor α(ERα)plays a significant role in the development of breast cancer.The main method used to determine the expression of ERα in breast cancer tissue is immunohistochemistry(IHC),but IHC results are affected by many factors.This study was designed to detect ERα expression in breast cancer tissues by RNAscope technique,and to compare the results of RNAscope with IHC results,then to further explore the clinical application of RNAscope technology.Methods: Collection of human breast cancer specimens(72 cases,47 cases of ERα positive and 25 cases of ERα negative).The collected samples were divided into three parts,one part for RNA extraction,RT-qPCR,one part for RNAscope in situ hybridization and IHC,one part for spare.Then the standard curve was constructed and the absolute quantitative RT-qPCR was performed.The results of absolute quantitative RT-qPCR and the results of RNAscope were compared with IHC.Results: The results of RT-qPCR and RNAscope of ERα-positive breast cancer tissues were significantly higher than those of ERα-negative breast cancer.The results of RT-qPCR were not consistent with the results of IHC,and the results of RNAscope were in good agreement with IHC results.In the analysis of inconsistent tissue specimens,we found that 3 IHC-based ERα negative but RNAscope-based positive cases would be diagnosed as positive after a second IHC test and 2 IHC-based ERα positive but RNAscope-based negative cases would be classified as negative,one IHC-based ERα negative but RNAscope-based positive case would be identified as negative.Conclusion:1.RNAscope results were in good agreement with IHC results,and RT-qPCR results were not obviously consistent with IHC results;and RNAscope operation is simpler than RT-qPCR operation,RNAscope also has visualization,compared with RT-qPCR,it has obvious advantages.2.In order to improve the diagnosis of ERα,RNAscope may be a complementary method for IHC. |
