Validation of a transfection protocol to study the functionality of small RNAs in the U-2932 cell line of diffuse large cell B lymphoma

Abstract

[eng] Transfection is a process that allows the entry of foreign material (DNA, RNA or drugs) into the cell. Reporting a problem in the transfection of cells in suspension and in particular the cells of haematological origin, which produces a difficulty in the progress of investigations. Especially in diffuse large cell lymphoma B (LDCGB), one of the most aggressive non-Hodgkin's lymphomas and common in the world. Among the main transfected compounds are siRNAs and miRNAs. The latter can influence the differentiation of B lymphocytes and oncogenesis in several types of lymphoma. This work aims to find the best method to transfect the U-2932 cell line (LDCGB) by comparing efficiency values, viability via MTT assay and functionality through miRNA mimics, among several transfection methods (electroporation, lipofection and Viromer). Among the main results, it was found that the efficiency achieved by electroporation was very low (<2%), accompanied by a lot of cell damage. While for Lipofectamine RNAiMAX average efficiency values between 30-40% were reported. For Viromer Green the highest transfection efficiency was found (> 75%) at 24 h and (> 90%) for incubation during 48 h. Viromer Green did not affect the cell viability, observed by the MTT and the tripan blue assays. Transfection with this agent was functional, evidenced by the decreased expression of protein kinase 9 (PTK9), used as a positive control. This work reports for the first time the transfection of this cell line with Lipofectamine RNAiMAX, and with the two types of Viromer.