Expression Pattern of Genes Associated With Tumor Microenvironment in Prostate Cancer

Abstract

Aim: To assess relative expression (RE) levels of CAF-, TAM-specific, immune defense-associated genes in prostate tumors and to show correlation of RE with clinical, pathological and molecular characteristics, with the aim to define clinically significant specific alterations in a gene expression pattern. Methods: RE of 23 genes was analyzed by a quantitative polymerase chain reaction in 37 freshly frozen samples of prostate cancer tissues of a different Gleason score (GS) and at various tumor stages, compared with RE in 37 paired conventionally normal prostate tissue (CNT) samples and 20 samples of prostate adenomas. Results: Differences in RE were shown for 11 genes out of 23 studied, when tumor samples were compared with corresponding CNTs. 7 genes, namely ACTA2, CXCL14, CTGF, THY1, FAP, CD163, CCL17 were upregulated in tumors. 4 genes, namely CCR4, NOS2A, MSMB, IL1R1 were downregulated in tumors. 14 genes demonstrated different RE in TNA at different stages: CXCL12, CXCL14, CTGF, FAP, HIF1A, THY1, CCL17, CCL22, CCR4, CD68, CD163, NOS2A, CTLA4, IL1R1. RE changes of 9 genes — CXCL12, CXCL14, HIF1A, CCR4, CCL17, NOS2A, CTLA4, IL1R1, IL2RA — were found in tumors with different GS. Moreover, 9 genes showed differences in RE in TNA, dependently on the presence or absence of the TMPRSS2/ERG fusion and 7 genes showed differences in RE of groups with differential PTEN expression. Significant correlations were calculated between RE of 9 genes in adenocarcinomas and the stage, and GS; also, between RE of 2 genes and the fusion presence; and between RE of 4 genes and PTEN expression. Conclusions: Several gene expression patterns were identified that correlated with the GS, stage and molecular characteristics of tumors, i.e. presence of the TMPRSS2/ERG fusion and alterations in PTEN expression. These expression patterns can be used for molecular profiling of prostate tumors, with the aim to develop personalized medicine approaches. However, the proposed profiling requires a more detailed analysis and a larger cohort of patients with prostate tumor.

The gene expression pattern as a tool for assessment of components of microenvironment and response to anti-cancer therapy of prostate tumors

“…An analysis of the gene expression pattern, assessed by a quantitative PCR method, is a powerful approach to solve important questions of modern molecular biology and medicine. Earlier, we used such analysis for the profiling of prostate cancers and creation of a diagnostic panel [1][2][3][4]. Next, we wanted to explore the possibility to use the gene expression analysis to predict the efficacy of anti-cancer therapy and to evaluate components of a tumor microenvironment.…”

“…A collection of prostate tissues. Samples of 37 prostate adenocarcinomas (PAC) of different stages and Gleason score and paired conventional normal tissues (CNT), and 20 benign prostate tumors (adenomas, AD) as well were collected as described earlier [2][3][4].…”

“…RE levels of investigated genes were detected by qPCR, using Maxima SYBR Green Master mix (Thermo Fisher Scientific, USA) with a Bio-Rad CFX96 Real-Time PCR Detection System (USA) under the following conditions: 95 °C -10 min, following 40 cycles of 95 °C -15 s, 60 °C -30 s, elongation 72 °C -30 s. Primers for genes were selected from a qPrimerDepot database (https://primerdepot.nci.nih.gov/). Four reference genes (TBP, HPRT, ALAS1, and TUBA1B) were used for the gene expression normalization [3,4]. Two main models for the calculation of RE levels were used, namely the Livak method 2 −∆Ct and the 2 −∆∆Ct calculation that show relative quantities and RE fold changes, accordingly [12].…”

“…The Kolmogorov-Smirnov test was used to analyze the normality of a distribution. The Kruskal-Wallis test and following the Dunn-Bonferoni post hoc test were performed to determine RE differences by multiple comparisons between experimental groups [1,4]. The Benjamini-Hochberg procedure with false discovery rate (FDR) 0.10-0.25 was used, when multiple comparisons were performed [13].…”

Approach to the Creation of a Gene Expression Panel for Prostate Cancer

“…Further studies of markers of CSI in order to assess the course of these diseases and the effectiveness of therapeutic measures are in the focus of interest [5]. The aim of our research was to study the dynamics of CSI markers in patients with coronary heart disease concurrent with AIT under the influence of polyphenol resveratrol [6].…”