Cholangiocarcinoma: tumoral suppressor miR-101-3P as a potential diagnostic and prognostic biomarker

• Grant number: 21/06315-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Effective date (Start): January 01, 2022
• Effective date (End): November 30, 2022
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Principal Investigator: Dorotéia Rossi Silva Souza
• Grantee: Sérgio Luís Ferreira Júnior
• Host Institution: Faculdade de Medicina de São José do Rio Preto (FAMERP). Secretaria de Desenvolvimento Econômico (São Paulo - Estado). São José do Rio Preto , SP, Brazil

Abstract

A malignant neoplasm that affects the bile ducts, cholangiocarcinoma (CCA) has a low survival rate due to the difficulties in detecting this type of cancer in its early stages. In this context, microRNAs (mRNAs) with inhibitory potential on tumor proliferation, such as mir-101-3p, is a potential therapeutic target involving diagnosis, prognosis, and treatment for CCA. Objectives - To evaluate the expression of miR-101-3p involved in oncogenesis in patients with CCA and its relationship with survival, lifestyle habits, and comorbidities. Methodology - 65 patients with CCA and 30 control subjects without clinical, biochemical, and histopathological signs of the disease, undergoing cholecystectomy, will be studied. The demographic and clinical profiles of both groups will be obtained by questionnaire and electronic medical records. For analysis of miR-101-3p expression, RNA will be extracted from tumor tissue embedded in paraffin (patients) and fresh cystic duct tissue (controls), using RealiaPrep" FFPE Total RNA Miniprep System (Promega) and triazole, respectively. Concentration and purity will be analyzed by NanoDrop-ND-1000 (Thermo Scientific) and the integrity of the sample in 1% agarose gel. The synthesis of complementary DNA (cDNA) will be performed via reverse transcription, using a specific kit (Taq Man - Applied Biosystems). Expression analysis of miR-101-3p will be performed by real-time reverse transcription-polymerase chain reaction (RT-qPCR). The transcription level will be calculated by the 2-””the Ct method. Data will be analyzed statistically, according to the characteristics of the variables, assuming a significance level for a value of P<0.05.(AU)