• An investigation on the role of ERBB4 and estrogen signaling pathway genes and microRNAs in papillary thyroid cancer
  • Javad Yaghmoorian Khojini,1 Mohammad Mokhtari,2,* Seyed Morteza Javadirad,3
    1. cellular and molecular biology and microbiology department, Faculty of biologic science and technologies , University of Isfahan
    2. Genetics Department, Faculty of biological science, Tarbiat Modares University
    3. cellular and molecular biology and microbiology department, faculty of biologic science and technologies , University of Isfahan


  • Introduction: Thyroid cancer is the most common endocrine malignancy worldwide and is among the main causes of cancer mortality. Surprisingly, prevalence of thyroid cancer in women is 3 times more than in men. It is proven that in cancers, expression of many genes in vital pathways alters, which can be the cause of the disease or the reason of its aggressive behaviors. ERBB4 pathway mainly plays its role in mitogenesis and differentiation, and aberrations and mutations in its genes associate with severe health issues such as cancers. On the other hand, estrogen dependent signaling pathways impact on metabolic activities, angiogenesis, and endocrine system function. Regulation disruptions of estrogen signaling pathways leads to severe disorders such as breast and ovarian cancer and metabolic disorders. Non-coding RNA based regulation is a newly discovered way of genome regulation which includes microRNAs, lncRNAs, siRNAs, and piRNAs. MicroRNAs or miRNAs are 21-23 nucleotide non-coding RNAs with clear function in cell division, differentiation, and apoptosis. The RNAs are responsible for post-transcriptional regulation, typically through sequence-specific interactions with 3’ untranslated region of mRNAs. In this project the expression level alterations of genes ERBB4 and estrogen signaling pathway genes and their respective microRNAs will be investigated in papillary thyroid carcinoma (PTC) patients.
  • Methods: In silico analysis Using GEO2R tool, PTC microarray data set (GSE3678) was analyzed and a set of genes with meaningfully altered expression was chosen. Then after, the network of these genes was drawn and the central nodes was depicted using STRING database. Using Reactome database, pathways holding the most gene hits was selected and involving genes was considered as the possible targets of expression analysis. Furthermore, possible microRNAs targeting each selected gene was predicted using mirwalk database. The most repeating microRNAs targeting multiple genes of each pathway was selected as the aim of expression studies. In vitro analysis The expression levels of selected genes and microRNAs was examined using qRT-PCR primers quantitatively. Moreover knockdown and Knock in of the mentioned genes was perform in cell culture and the behavior of the cells was analyzed. SiRNA transfection of selected microRNAs was performed and the regulation of genes by respective microRNAs was confirmed using luciferase assay and western blot.
  • Results: In silco results Out of 25000 genes 163 genes with meaning full expression alteration was selected and after visualization of their network and related pathways, ERBB4, ERBB3, GABRB2, ELMO1, WWOX, and LRRK2 Genes was selected to be studied in ERBB4 signaling pathway. Also ERBB4, TFF3, TGFA, CITED1, and JUN genes of the estrogen signaling pathway was chosen to be study. To investigate the regulatory roles of microRNAs on these pathways in PTC hsa-miR-204-5p, hsa-miR-335-5p, hsa-miR-211-5p, hsa-miR-1285-3p, hsa-miR-612, hsa-miR-205-5p was considered for the expression analysis studies.
  • Conclusion: Laboratory investigation of this study is to be done and hopefully, the results can be a great help in detection and treatment of patients suffering this catastrophic disease, as well as base for future researchers.
  • Keywords: Papillary thyroid cancer, microRNA, qRT-PCR, western blotting