Повышенная экспрессия генов AKR1C2 и AKR1B1 связана с развитием резистентности к доцетакселу при раке предстательной железы.

Prostate cancer (PC) is a highly prevalent and socially impactful malignancy that affects males globally. In Russia, the incidence of PC is also notably high, with 40,785 newly reported cases in 2017

PC3 cells were cultured in RPMI-1640 medium at a temperature of 37C with a 5% CO2. To generate resistant cell sublines, we employed a gradual increase in DoX concentration in the medium. Cells that survived at a DoX concentration of 10 nM were deemed resistant. We performed RNA extraction using the RNeasy Micro Kit (Qiagen) and library preparation using the KAPA RNA HyperPrep Kit with RiboErase (Roche). Next-generation sequencing (NGS) was conducted on the NextSeq 2000 instrument. Analysis of differential gene expression was carried out utilizing the edgeR in the R. To perform reverse transcription, we utilized the Mint kit (Evrogen) and conducted quantitative polymerase chain reaction (qPCR) on the Applied Biosystems 7500 instrument with three technical replicates. The HPRT1 gene was used for normalization. The relative mRNA expression level for each comparison was determined using the dCT method. Statistical significance of gene expression differences was assessed with a criterion of p<0,05.

The key findings related to the varying expression of AkRs genes in different experimental cell groups during DoX treatment are summarized in Table 1. When comparing the DoX 4 nM and 10 nM groups, the results showed statistically significant findings for the AKR1C2 and AKR1B10 genes. Importantly, the differential expression of AKR1C2 and AKR1B10 exhibited a remarkable increase in DoX resistant cells compared to cells treated with DoX initially. Specifically, AKR1C2 expression showed a 195-fold increase, while AKR1B10 expression showed a 10.63-fold increase. Furthermore, in the comparison between resistant cells and the control group, the expression of AKR1C2 was enhanced by a 3.2-fold increase, while AKR1B10 expression showed a 2.19-fold increase. Additionally, when comparing the DoX 8nM and DoX 10nM experimental cell groups, there was a notable and statistically significant upregulation of AKR1B10 and AKR1C2 expression. Notably, the differential expression of AKR1C2 showed a remarkable 10-fold increase in this comparison.

The evaluation focused on the changes in gene expression levels of AKR1C2 and AKR1B10 after being exposed to DoX treatment. The findings demonstrated a significant rise in AKR1C2 expression within the DoX 10 nM group, as compared to both the DoX 4 nM group (p = 0.001) and the DoX 8 nM group (p = 0.002) (Fig. 1). In particular, the AKR1C2 expression in the DoX 10nM group showed a significant 13.82-fold rise compared to the DoX 4nM group. Moreover, there was a notable 4.88-fold increase in AKR1C2 expression in the DoX 10nM group in comparison to the DoX 8nM group.

Figure 1 - Boxplots of relative expression of AKR1C2 mRNA levels among different experimental groups

Note: the y-axis represents the 2-ΔCT values

DOI:10.18454/jbg.2023.22.10.2

Additionally, the findings demonstrated a notable increase in AKR1B10 expression within the DoX 10nM group when compared to both the DoX 4nM group (p = 0.0002) and the DoX 8nM group (p = 0.02) (Fig. 2). In the DoX 10nM group, the expression of AKR1B10 showed a significant 6.3-fold increase compared to the DoX 4nM group. Furthermore, a notable 1.84-fold increase in AKR1B10 expression was observed in the DoX 10nM group compared to the DoX 8nM group.

Figure 2 - Boxplots of relative expression of AKR1B10 mRNA levels among different experimental groups of cells

Note: the y-axis represents the 2-ΔCT values

DOI:10.18454/jbg.2023.22.10.3

As a result, qPCR was utilized to quantitatively examine AKR1C2 and AKR1B10 gene transcripts, and it confirmed significant differences between the groups under investigation. This further validated the findings obtained from NGS and demonstrated a noteworthy rise in the expression levels of two AkRs members, AKR1C2 and AKR1B10, during the progression of DoX resistance in crPC.

Reviewing the literature, it has been observed that the overexpression of the AKR1B10 gene is frequently found in different tumor types and may be linked tothe disease

Chemotherapy resistance continues to be a major obstacle in the clinical use of these drugs. Uncovering potential therapeutic targets involved in resistance could greatly enhance the efficacy of these agents and ultimately enhance patient survival rates. In the study conducted on PC cells as a model, the researchers examined the differential gene expression of AkRs during the development of resistance to the taxane drug, DoX. The expression levels of these genes have been found to be higher in different tumor types, suggesting their potential involvement in chemoresistance. The study results indicate a strong statistical correlation between the overexpression of two specific genes, AKR1C2 and AKR1B10, and the development of resistance to DoX in PC3 cells.