Optimization of multiplex RT-PCR for selected isoforms of metallothionein genes and influence of cisplatin on prostatic cell lines

Abstract

This study deals with the issue of metallothioneins as potential tumor markers using the multiplex reverse transcription polymerase chain reaction (RT-PCR) method, for which specific primers were designed, with the subsequent sequencing of their products. The following step involved a study of expression using the PCR method for selected MT1A, MT2A and MT3 genes after treating cell lines with cisplatin. Experiments were carried out on PNT1A, LNCaP and DU145 prostatic cell lines. Despite a very high similarity of the individual isoforms of metallothioneins, a triplex of three genes for two cell lines, PNT1A and DU145, was created. Upon a statistical evaluation of data obtained via the method of polymerase chain reaction in real time, it was found that, after the application of cisplatin, there was an increase in metallothionein expression.This study deals with the issue of metallothioneins as potential tumor markers using the multiplex reverse transcription polymerase chain reaction (RT-PCR) method, for which specific primers were designed, with the subsequent sequencing of their products. The following step involved a study of expression using the PCR method for selected MT1A, MT2A and MT3 genes after treating cell lines with cisplatin. Experiments were carried out on PNT1A, LNCaP and DU145 prostatic cell lines. Despite a very high similarity of the individual isoforms of metallothioneins, a triplex of three genes for two cell lines, PNT1A and DU145, was created. Upon a statistical evaluation of data obtained via the method of polymerase chain reaction in real time, it was found that, after the application of cisplatin, there was an increase in metallothionein expression.