Long noncoding RNA HOTTIP is associated with male infertility and promotes testicular embryonal carcinoma cell proliferation

Abstract

Background It has been proposed that lncRNAs, widely transcribed from genomes, play pivotal regulatory roles in a variety of biological processes, but their function in regulating spermatogenesis in human males is rarely reported. Methods QRT‐PCR was adopted to detect HOTTIP expression level in testicular tissues from hypospermatogenesis (Hypo) patients or controls. The proliferation levels of NT2 and 293T were measured via CCK‐8 and EdU detection. Meanwhile, luciferase reporter gene assay and bioinformatics analysis were carried out to identify a target of HOTTIP. Additionally, the underlying mechanism of HOTTIP’s function was investigated using western blotting and RIP analysis. Results The research results manifested that the expression of HOTTIP in testicular tissues from Hypo patients was prominently reduced in comparison with that in control testicular tissues. Interestingly, it was noted that HOTTIP exhibited a high expression in testicular embryonal carcinoma cell line NT2 compared with that in normal control cell line 293T. It was denoted in cell function evaluation that cell proliferation was impeded by downregulated HOTTIP but evidently stimulated by overexpressed HOTTIP. Moreover, HOTTIP was capable of positively modulating HOXA13 expression via the competitive binding to miR‐128‐3p. Conclusion Therefore, HOTTIP acting as ceRNAs to promote testicular embryonal carcinoma cell proliferation.

Transcript Isoforms of SLC7A11-AS1 Are Associated With Varicocele-Related Male Infertility

“…Since the role of SLC7A11-AS1 was so far not evaluated in relation to oxidative stress, we used the advantage of NT2 and NCCIT cell lines to assess the function of SLC7A11-AS1 in the regulation of its neighboring protein-coding gene, SLC7A11. It is noteworthy that these two cell lines (NT2 and NCCIT) as testicular germ cell carcinoma cell lines have been previously used for functional analysis of lncRNAs in male infertility (Lü et al, 2015;Su et al, 2019).…”

Long Non-Coding RNAs: Role in Testicular Cancers

“…Both SPRY4-IT1 and HOTTIP were shown to increase testicular cancer proliferation ( 100 , 102 ), so their inhibition or downregulation could lead to a decrease of tumor aggressiveness and size. This would make adjuvant chemotherapy dispensable for localized masses, which could be more easily removed via surgical excision.…”

“…The oncogenic role of HOTTIP is not solely mediated by WDR5 action, but also by the interaction with several different chromatin regulators, oncogenic pathways, and miRNAs depending on the cancerous setting in which is involved ( 102 ). In testicular embryonal carcinoma, HOTTIP was described to promote cell proliferation via the competitive binding to miR‐128‐3p ( 102 ).…”

“…In fact, HOTTIP was found upregulated in testicular embryonal carcinoma cells compared to the control, and its knock-out in those cells resulted in a decrease of the proliferative rate ( 102 ). Bioinformatic considerations based on HOTTIP structure, later validated in vitro , proved that its effect on cell proliferation was mediated by miR‐128‐3p, that is sponged by HOTTIP ( 148 ).…”

“…Furthermore, HOTTIP /miR‐128‐3p pathway was found to positively regulate HOXA13 expression, which is quite a canonical downstream effector of HOTTIP in cancerogenic settings ( 102 , 148 ). In fact, overexpression of HOXA13 in response to HOTTIP oncogenic action was described in small cell lung cancer, gastric cancer, pancreatic, hepatocellular and esophageal squamous cell carcinoma, and in some of these setting its expression alteration is triggered by the inhibition of a miRNA ( 148 ).…”