Clinical value of detecting seminal plasma levels of miR-135a-5p and miR-135b-5p in male patients with infertility / 临床检验杂志

Objective To examine the expression pattern of miR-135a-5p and miR-135b-5p in seminal plasma as well as the clinical value of their variation in the male patients with infertility.Methods The clinical samples of 5 kinds,including serum,morning urine,pleural effusion,cerebrospinal fluid and seminal plasma,were collected from 30 patients respectively in the Department of Clini-cal Laboratory,Jiangsu Province Hospital of Chinese Medicine from January 2019 to December 2021.In addition,the frozen seminal plasma samples from the patients with non-obstructive azoospermia,asthenozoospermia of 54 cases for each illness statuses,and 54 healthy controls with fertile ability from 2010 to 2019 in Jiangsu Province Hospital of Chinese Medicine and Jinling Hospital were also selected.Fluorescent quantitative RT-qPCR analyses were applied to measure the expression levels of miR-135a-5p and miR-135b-5p in seminal plasma.Receiver operating characteristic curve(ROC)was used to construct the area under the curve(AUCROC)to evalu-ate the ability for discriminating infertility males from healthy controls.Correlation analyses were performed to explore the correlations between the levels of miR-135a-5p and miR-135b-5p in seminal plasma and other semen parameters.Results The concentrations of miR-135a-5p and miR-135b-5p in the samples from five different kinds of body fluid showed significantly difference(H=64.88,P＜0.01 and H=64.7,P＜0.01,respectively).The highest levels were found in serum and seminal plasma,and the lowest levels were in u-rine.The concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were also showed marked difference among the azoosper-mia group,asthenozoospermia group and healthy fertile control group(H=32.14,P＜0.01 and H=21.37,P＜0.01,respectively).Compared to the fertile control group,the levels of miR-135a-5p and miR-135b-5p in seminal plasma were significantly downregulated in azoospermia patients(U=687,P＜0.01 and U=843,P＜0.01,respectively).The contents of the two miRNAs showed no statistical-ly difference between asthenozoospermia patients and healthy fertile controls,but showed markedly difference between the two patients groups(U=635,P＜0.01 and U=779,P＜0.01,respectively).Receiver operating characteristic curve(ROC)analyses results re-vealed that levels of miR-135a-5p and miR-135b-5p in seminal plasma could successfully discriminate the azoospermia patients from fertile controls and asthenozoospermia patients with the AUCROC of 0.764(95％CI:0.675 to 0.854),0.711(95％CI:0.612 to 0.810)and 0.782(95％CI:0.695 to 0.869),and 0.733(95％CI:0.637 to 0.829),respectively.Correlation analyses demonstrated that the concentrations of miR-135a-5p and miR-135b-5p in seminal plasma were significantly associated with sperm concentration,total sperm motility,and percentages of progressive sperm and non-progressive sperm(all with the P-value＜0.01).Conclusion miR-135a-5p and miR-135b-5p are the highly expressed miRNA in seminal plasma,and significantly decreased in azoospermia patients.Both of them may play pivot roles in male infertility.

Effects of miR-135a-5p on the expression of insulin-like factor 3 and testosterone in flutamide-induced cryptorchidism mouse / 安徽医科大学学报

Objective @# To investigate the effect of miR-135a-5p on the expression levels of insulin-like factor 3 (INSL3) and testosterone in the testicular tissues of flutamide-induced cryptorchidism mouse．@*Methods @# A model of flutamide-induced cryptorchidism in mouse was constructed，and the experiment was divided into normal control group，flutamide group，flutamide + miR-135a-5p knockdown group and flutamide + miR-135a-5p overexpression group. RT-qPCR was used to detect the expression levels of miR-135a-5p and INSL3 mRNA．Western blot was em- ployed to assess the protein expression level of INSL3．ELISA was performed to measure the expression level of tes- tosterone. @*Results @#The expression levels of miR-135a-5p，INSL3 mRNA and protein and testosterone were significantly down-regulated in the testis of cryptorchid mice by flutamide (P＜0. 05) ．Knockdown of miR-135a-5p could downregulate the expression of INSL3 mRNA，INSL3 protein and testosterone (P ＜0. 01 ) ，while overexpression of miR-135a-5p had the opposite result．@*Conclusion @#miR-135a-5p decreased in flutamide-induced cryptorchidism mouse testicular tissues，and overexpression of miR-135a-5p could restore the expression levels of INSL3 and testosterone.

MiR-135a-5p suppresses breast cancer cell proliferation, migration, and invasion by regulating BAG3

Abstract Background MicroRNAs (miRNAs) are involved in the progression of diverse human cancers. This work aimed to delve into how microRNA-135a-5p (miR-135a-5p) affects the biological behaviors of Breast Cancer (BC) cells. Methods Gene Expression Omnibus (GEO) datasets were used to analyze the expression differences of miR-135a-5p in cancer tissues of BC patients. Quantitative real-time PCR and western blot were conducted to detect miR-135a-5p and Bcl-2 Associated Athanogene (BAG3) expression levels in BC tissues and cells, respectively. The proliferation, migration, invasion, and cell cycle of BC cells were detected by cell counting kit-8 assay, BrdU assay, wound healing assay, transwell assay, and flow cytometry. The targeted relationship between miR-135a-5p and BAG3 mRNA 3′UTR predicted by bioinformatics was further testified by a dual-luciferase reporter gene assay. Pearson's correlation analysis was adopted to analyze the correlation between miR-135a-5p expression and BAG3 expression. The downstream pathways of BAG3 were analyzed by the LinkedOmics database. Results MiR-135a-5p was significantly down-regulated and BAG3 expression was significantly raised in BC tissues. MiR-135a-5p overexpression repressed the viability, migration and invasion of BC cells, and blocked cell cycle progression in G0/G1 phase while inhibiting miR-135a-5p worked oppositely. BAG3 was verified as a target of miR-135a-5p. Overexpression of BAG3 reversed the impacts of miR-135a-5p on the malignant biological behaviors of BC cells. The high expression of BAG3 was associated with the activation of the cell cycle, mTOR and TGF-β signaling pathways. Conclusion MiR-135a-5p regulates BAG3 to repress the growth, migration, invasion, and cell cycle progression of BC cells.

Expression and Significance of miR?135a?5p in Endometrial Carcinoma / 中国医科大学学报

Objective To investigate the expression and significance of miR?135a?5p in endometrial carcinoma and to correlate expression levels with clinicopathological features. Methods Fifty five endometrial carcinoma samples and thirty normal endometrial tissue samples were surgically obtained,and the expression of miR?135a?5p was quantitated by real?time polymerase chain reaction. The relationship between the expression level of miR?135a?5p and clinicopathological data of individual patients was analyzed. Results The expression of miR?135a?5p was lower in the endo?metrial carcinoma group than in the normal endometrial group(P=0.021),and correlated significantly with the histologic grade of the endometrial neoplasm(P=0.008). Conclusion The expression of miR?135a?5p is lower in endometrial carcinoma than in normal endometrial tissue. In en?dometrial carcinoma,the miR?135a?5p expression status has a statistically significant relationship with the histological characteristics of the tumor.