Microarray and in silico analysis of DNA repair genes between human testis of patients with nonobstructive azoospermia and normal cells

Abstract

DNA repair processes are critical to maintaining genomic integrity. As a result, dysregulation of repair genes is likely to be linked with health implications, such as an increased prevalence of infertility and an accelerated rate of aging. We evaluated all the DNA repair genes (322 genes) by microarray. This study has provided insight into the connection between DNA repair genes, including RAD23B, OBFC2A, PMS1, UBE2V1, ERCC5, SMUG1, RFC4, PMS2L5, MMS19, SHFM1, INO80, PMS2L1, CHEK2, TRIP13, and POLD4. The microarray analysis of six human cases with different nonobstructive azoospermia revealed that RAD23B, OBFC2A, PMS1, UBE2V1, ERCC5, SMUG1, RFC4, PMS2L5, MMS19, SHFM1, and INO80 were upregulated, and expression of PMS2L1, CHEK2, TRIP13, and POLD4 was downregulated versus the normal case. For this purpose, Enrich Shiny GO, STRING, and Cytoscape online evaluation was applied to predict proteins’ functional and molecular interactions and then performed to recognize the master pathways. Functional enrichment analysis revealed that the biological process (BP) terms “base‐excision repair, AP site formation,” “nucleotide‐excision repair, DNA gap filling,” and “nucleotide‐excision repair, preincision complex assembly” was significantly overexpressed in upregulated differentially expressed genes (DEGs). BP analysis of downregulated DEGs highlighted “histone phosphorylation,” “DNA damage response, detection DNA response,” “mitotic cell cycle checkpoint signaling,“ and “double‐strand break repair.” Overrepresented molecular function (MF) terms in upregulated DEGs included “Oxidized base lesion DNA N‐glycosylase activity,” “uracil DNA N‐glycosylase activity,” “bubble DNA binding” and “DNA clamp loader activity.” Interestingly, MF investigation of downregulated DEGs showed overexpression in “heterotrimeric G‐protein complex,” “5′‐deoxyribose‐5‐phosphate lyase activity,” “minor groove of adenine‐thymine‐rich DNA binding,” and “histone kinase activity.” Our findings suggest that these genes and their interacting hub proteins could help determine the pathophysiology of germ cell abnormalities and infertility.

Whole Exome Sequencing and In Silico Analysis of Human Sertoli in Patients with Non-Obstructive Azoospermia

“…The attached cells were selected for Microarray studies. Our experiments selected highly pure Vimentin+, Sox9+, Gata4+,Vasa-, and UTF1-human Sertoli cell cultures [ 59 ].…”

Undifferentiated and Differentiated Spermatogonial Stem Cells

“…A high quality and relevance of gene prioritization can be seen in this network, which contains five of the eight canonical markers. As well as finding novel regulators of SSC self-renewal, we suggest the refined network could be used to identify target genes for male infertility and testicular cancer treatment [16,29,[31][32][33]. DOI: http://dx.doi.org/10.5772/intechopen.112964…”

Functions and mechanism of noncoding RNA in regulation and differentiation of male mammalian reproduction

“…93 It is also plausible that TEs temporarily relaxed at the time when MILI vanished during the prelepotonema and leptonema phases. [93][94][95][96][97] The TE promoters in the nucleus may also be methylated as a result of MIWI2. The L1 element may be derepressed and its expression level can be increased without having any impact on mouse spermatogenesis when two piRNAs, NCT1/2, which are transiently expressed from one cluster on chromosome 2, lose their function.…”

“…In fact, CpG methylation is significantly impacted by Miwi2 and Mili mutations for both L1 and IAP retrotransposons 93 . It is also plausible that TEs temporarily relaxed at the time when MILI vanished during the prelepotonema and leptonema phases 93–97 …”