Depletion of HMGB2 causes seminiferous tubule atrophy via aberrant expression of androgen and estrogen receptors in mouse testis

Abstract High-mobility group box 2 (HMGB2), a chromatin-associated protein that interacts with DNA, is implicated in multiple biological processes, including gene transcription, replication, and repair. HMGB2 is expressed in several tissues, including the testis; however, its functional role is largely unknown. Here, we elucidated the role of HMGB2 in spermatogenesis using HMGB2 knock-out (KO) mice. Paraffin-embedded testicular tissues were obtained from 8-week-old and 1-year-old wild-type and KO mice. Testis weight and number of seminiferous tubules were decreased, whereas atrophic tubules were increased in HMGB2-depleted mice. Immunohistochemistry revealed that atrophic tubules contained Sertoli cells, but not germ cells. Moreover, decreased cell proliferation and increased apoptosis were demonstrated in HMGB2-depleted mouse testis. To elucidate the cause of tubule atrophy, we examined the expression of androgen and estrogen receptors (AR, ERs, respectively), and the results indicated aberrant expression of AR and ERα in Sertoli and Leydig cells. Southwestern histochemistry detected decreased estrogen response element–binding sites in HMGB2-depleted mouse testis. Expression of HMGB1, which has highly similar structure and function as HMGB2, was examined by immunohistochemistry and western blotting, which indicated increased expression in aged HMGB2 KO mouse testis, especially in spermatocytes. These findings indicate a compensatory increase in HMGB1 expression in HMGB2 KO mouse testis. In summary, depletion of HMGB2 induced aberrant expression of AR and ERα, leading to decreased germ cell proliferation and increased apoptosis that resulted in focal seminiferous tubule atrophy.

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Relationship between testicular morphology and sperm production following ischaemia in the ram

Reproduction Fertility and Development ◽

10.1071/rd9950119 ◽

1995 ◽

Vol 7 (1) ◽

pp. 119 ◽

Cited By ~ 5

Author(s):

CM Markey ◽

AM Jequier ◽

GT Meyer ◽

GB Martin

Keyword(s):

Morphological Changes ◽

Sperm Concentration ◽

Testicular Biopsy ◽

Seminiferous Tubules ◽

Testis Weight ◽

Infertile Men ◽

Testicular Morphology ◽

And Function ◽

The Relationship ◽

Spermatogenic Epithelium

Arteriosclerosis was induced in the internal spermatic artery of rams to determine if this condition is implicated in the aetiology of testicular pathology which causes male infertility. Data were collected on sperm concentration and motility for 56 days following surgery to provide an index of testicular function. Testes were then weighed and a testicular biopsy score count was performed on histological sections to assess spermatogenic potential of seminiferous tubules. Vascular disturbance caused focal damage of the seminiferous epithelium, similar to that seen among infertile men, and a reduction in ejaculate volume, sperm concentration and sperm motility. Sperm concentration decreased following ischaemia yet was maintained to some degree by a germ-cell depleted spermatogenic epithelium. Normal testicular morphology was maintained above a testis weight of about 120 g (for an individual testis), but below this threshold spermatogenesis was severely impaired. In conclusion, these data have provided information on the relationship between testicular morphology and function following ischaemia in the ram. Furthermore, the morphological changes induced in the testis were similar to those seen among infertile men and, by their focal nature, could explain the distinction between oligozoospermia and azoospermia in men exhibiting spermatogenic arrest.

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Secretion of transferrin in ovine seminiferous tubule cell cultures in response to FSH: influence of breed, season of birth and age of lambs

Reproduction Fertility and Development ◽

10.1071/rd9930083 ◽

1993 ◽

Vol 5 (1) ◽

pp. 83 ◽

Cited By ~ 1

Author(s):

C Monet-Kuntz ◽

I Fontaine

Keyword(s):

Cell Cultures ◽

Dose Response ◽

Sertoli Cells ◽

Seminiferous Tubule ◽

Seminiferous Tubules ◽

Testis Weight ◽

Season Of Birth ◽

Tubule Cell ◽

Response Curves ◽

Testicular Weight

The response of lamb Sertoli cells to follicle stimulating hormone (FSH) was investigated by measuring transferrin secretion in seminiferous tubule cell cultures throughout the non-pubertal and the prepubertal periods. Cells could be cultured from birth until they attained a testicular weight of 19 g. The characteristics of individual dose-response curves were compared according to the breed, season of birth and testicular weight of the lambs. At the same season of birth and within a given testis weight range, dose-response curves of Romanov and Ile-de-France lambs were similar. Within a given testis weight range, spring-born animals exhibited a higher maximal transferrin secretion than autumn-born lambs, but the ED50 was similar. The main factor of variation of the dose-response curve parameters was the testicular weight of the lambs: the amplitude of FSH response increased 3-fold from a testicular weight of 6 g onwards, i.e. from the appearance of spermatogonia in seminiferous tubules. The ED50 increased 5-fold from 11 g onwards, i.e. from the beginning of the prepubertal period. Thus, Sertoli cells become less sensitive to FSH as spermatogenesis develops in seminiferous tubules. This phenomenon is largely the result of higher phosphodiesterase activity and is greatly reduced by 1-methyl-3-isobutyl-xanthine (MIX).

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Effects of a simulated microgravity model on cell structure and function in rat testis and epididymis

Journal of Applied Physiology ◽

10.1152/jappl.1992.72.2.748 ◽

1992 ◽

Vol 72 (2) ◽

pp. 748-759 ◽

Cited By ~ 14

Author(s):

J. A. Hadley ◽

J. C. Hall ◽

A. O'Brien ◽

R. Ball

Keyword(s):

Cell Structure ◽

Abdominal Cavity ◽

Structure And Function ◽

Male Rats ◽

Serum Prolactin ◽

Seminiferous Tubules ◽

Testis Weight ◽

Testosterone Levels ◽

Multinucleated Cells ◽

And Function

A tail-suspension (TS) rat model used to simulate microgravity was tested for its effects on the anatomy, cell structure, and function of the testis and epididymis in sexually mature male rats. Rats suspended for 7 days without inguinal canal ligation exhibited a significant (P less than or equal to 0.05) reduction in testis weight compared with controls (1.55 +/- 0.04 to 1.1 +/- 0.02 g). Except for the liver, epididymis, and adrenals of TS rats and TS rats allowed to recover for 7 days, no significant (P less than or equal to 0.05) change was observed in the weight of other body and accessory sex organs. A histological examination of the testes and epididymides of model animals revealed disorganized seminiferous tubules and accumulation of large multinucleated cells and spermatids in the lumen of the epididymis. A significant (P less than or equal to 0.05) increase in serum luteinizing hormone (53.1 +/- 6.7 to 66.2 +/- 10.1 ng/ml) and follicle-stimulating hormone (257 +/- 25 to 305 +/- 38 ng/ml) was observed in TS nonligated rats, whereas serum prolactin and testosterone levels were observed to decline from 8.3 +/- 1.3 to 5.1 +/- 0.29 and 7.1 +/- 1.3 to 3.8 +/- 0.25 ng/ml, respectively. Decreases in testis protein content and testosterone levels of the testis, interstitial fluid, and epididymis were also observed in model animals. These data demonstrate that the suspension procedure used in the National Aeronautics and Space Administration TS model results in the testis and epididymis translocating into the abdominal cavity, causing cellular degeneration and organ dysfunction.

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Wild boars (Sus scrofa scrofa) seminiferous tubules morphometry

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132006000600008 ◽

2006 ◽

Vol 49 (5) ◽

pp. 739-745 ◽

Cited By ~ 10

Author(s):

Deiler Sampaio Costa ◽

José Frederico Straggiotti Silva

Keyword(s):

Sertoli Cell ◽

Sertoli Cells ◽

Seminiferous Tubule ◽

Sus Scrofa ◽

Seminiferous Tubules ◽

Efficiency Coefficient ◽

Wild Boars ◽

Intrinsic Efficiency ◽

And Function ◽

Tubular Diameter

The aim of this data was to analyze morphology and function of the seminiferous tubule in adult wild boars. Testes removed by unilateral castration of five animals were used. The testicular parenchyma was composed by 82.1±2.2% of seminiferous tubule and 17.9±2.2% of intertubular tissue. The tubular diameter was 249.2±33.0 µm and the seminiferous tubule lenght per gram of testis was 19.3±4.9m. The spermatogonial mitoses efficiency coefficient, meiotic index and spermatogenesis efficiency were 10.34, 2.71 and 30.5 respectively. Each Sertoli cell supported about 13 germinatives cells. The hystometric parameters studied were very similar to those related for domestic boars, however, the wild boars intrinsic efficiency of spermatogenesis and Sertoli cells indexes were smaller than in domestic boars.

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Tead1 Reciprocally Regulates Adult β-Cell Proliferation and Function to Maintain Glucose Homeostasis

SSRN Electronic Journal ◽

10.2139/ssrn.3704009 ◽

2020 ◽

Author(s):

Jeongkyung Lee ◽

Ruya Liu ◽

Byung S. Kim ◽

Yiqun Zhang ◽

Feng Li ◽

...

Keyword(s):

Cell Proliferation ◽

Glucose Homeostasis ◽

Β Cell ◽

And Function

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Antigene and Antiproliferative Effects of Triplex-Forming Oligonucleotide (TFO) Targeted on hmgb1 Gene in Human Hepatoma Cells

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200619170438 ◽

2020 ◽

Vol 20 (16) ◽

pp. 1943-1955

Author(s):

Neelam Lohani ◽

Moganty R. Rajeswari

Keyword(s):

Apoptotic Cell ◽

Isothermal Calorimetry ◽

High Mobility ◽

Rational Drug Design ◽

Human Hepatoma Cells ◽

Aberrant Expression ◽

Bioinformatic Tools ◽

Hmgb1 Expression ◽

Dna Triplex ◽

Triplex Forming Oligonucleotide

Background: The high mobility group box 1 (hmgb1) is one of the frequently over-expressed genes whose aberrant expression is reported in a number of human cancers. Various strategies are underway to inhibit hmgb1 expression in cancer cells having considerable therapeutic value. Objective: The present work involves selective transcriptional inhibition of the hmgb1 gene using selective DNA triplex structure-based gene technology. Here, the promoter region of the hmgb1 gene at position (-183 to -165) from the transcription start site as a target was selected using bioinformatic tools. Methods: The DNA triplex formation by the DNA of the target gene and TFO was confirmed using UV absorption spectroscopy, Circular Dichroism, and Isothermal Calorimetry. Results: Treatment of HepG2 cell with specific Triplex-forming Oligonucleotide significantly downregulated HMGB1 expression level at mRNA and protein levels by 50%, while the classical anticancer drugs, actinomycin/ adriamycin as positive controls showed 65% and the combination of TFO and drug decreased by 70%. The anti-proliferative effects of TFO correlated well with the fact of accumulation of cells in the Go phase and apoptotic cell death. Further, the binding of anti-cancer drugs to hmgb1 is stronger in DNA triplex state as compared to hmgb1 alone, suggesting the combination therapy as a better option. Conclusion: Therefore, the ability of hmgb1 targeted triplex-forming oligonucleotide in combination with triplex selective anticancer drug holds promise in the treatment of malignancies associated with hmgb1 overexpression. The result obtained may open up new vistas to provide a basis for the rational drug design and searching for high-affinity ligands with a high triplex selectivity.

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KLF7/VPS35 axis contributes to hepatocellular carcinoma progression through CCDC85C-activated β-catenin pathway

Cell & Bioscience ◽

10.1186/s13578-021-00585-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Yarong Guo ◽

Bao Chai ◽

Junmei Jia ◽

Mudan Yang ◽

Yanjun Li ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Tumor Growth ◽

Luciferase Reporter ◽

Aberrant Expression ◽

Proliferation And Invasion ◽

Hcc Cells

Abstract Objective Dysregulation of KLF7 participates in the development of various cancers, but it is unclear whether there is a link between HCC and aberrant expression of KLF7. The aim of this study was to investigate the role of KLF7 in proliferation and migration of hepatocellular carcinoma (HCC) cells. Methods CCK8, colony growth, transwell, cell cycle analysis and apoptosis detection were performed to explore the effect of KLF7, VPS35 and Ccdc85c on cell function in vitro. Xenografted tumor growth was used to assess in vivo role of KLF7. Chip-qPCR and luciferase reporter assays were applied to check whether KLF7 regulated VPS35 at transcriptional manner. Co-IP assay was performed to detect the interaction between VPS35 and Ccdc85c. Immunohistochemical staining and qRT-PCR analysis were performed in human HCC sampels to study the clinical significance of KLF7, VPS35 and β-catenin. Results Firstly, KLF7 was highly expressed in human HCC samples and correlated with patients’ differentiation and metastasis status. KLF7 overexpression contributed to cell proliferation and invasion of HCC cells in vitro and in vivo. KLF7 transcriptional activation of VPS35 was necessary for HCC tumor growth and metastasis. Further, co-IP studies revealed that VPS35 could interact with Ccdc85c in HCC cells. Rescue assay confirmed that overexpression of VPS35 and knockdown of Ccdc85c abolished the VPS35-medicated promotion effect on cell proliferation and invasion. Finally, KLF7/VPS35 axis regulated Ccdc85c, which involved in activation of β-catenin signaling pathway, confirmed using β-catenin inhibitor, GK974. Functional studies suggested that downregulation of Ccdc85c partly reversed the capacity of cell proliferation and invasion in HCC cells, which was regulated by VPS35 upregulation. Lastly, there was a positive correlation among KLF7, VPS35 and active-β-catenin in human HCC patients. Conclusion We demonstrated that KLF7/VPS35 axis promoted HCC cell progression by activating Ccdc85c-medicated β-catenin pathway. Targeting this signal axis might be a potential treatment strategy for HCC.

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Deficiency of the onco-miRNA cluster, miR-106b∼25, causes oligozoospermia and the cooperative action of miR-106b∼25 and miR-17∼92 is required to maintain male fertility

MHR Basic science of reproductive medicine ◽

10.1093/molehr/gaaa027 ◽

2020 ◽

Vol 26 (6) ◽

pp. 389-401

Author(s):

Alicia Hurtado ◽

Rogelio Palomino ◽

Ina Georg ◽

Miguel Lao ◽

Francisca M Real ◽

...

Keyword(s):

Male Fertility ◽

Molecular Mechanisms ◽

Mirna Cluster ◽

Knock Out ◽

Cooperative Action ◽

Mouse Mutants ◽

New Genes ◽

Mirna Clusters ◽

Testicular Histology ◽

And Function

Abstract The identification of new genes involved in sexual development and gonadal function as potential candidates causing male infertility is important for both diagnostic and therapeutic purposes. Deficiency of the onco-miRNA cluster miR-17∼92 has been shown to disrupt spermatogenesis, whereas mutations in its paralog cluster, miR-106b∼25, that is expressed in the same cells, were reported to have no effect on testis development and function. The aim of this work is to determine the role of these two miRNA clusters in spermatogenesis and male fertility. For this, we analyzed miR-106b∼25 and miR-17∼92 single and double mouse mutants and compared them to control mice. We found that miR-106b∼25 knock out testes show reduced size, oligozoospermia and altered spermatogenesis. Transcriptomic analysis showed that multiple molecular pathways are deregulated in these mutant testes. Nevertheless, mutant males conserved normal fertility even when early spermatogenesis and other functions were disrupted. In contrast, miR-17∼92+/−; miR-106b∼25−/− double mutants showed severely disrupted testicular histology and significantly reduced fertility. Our results indicate that miR-106b∼25 and miR-17∼92 ensure accurate gene expression levels in the adult testis, keeping them within the required thresholds. They play a crucial role in testis homeostasis and are required to maintain male fertility. Hence, we have identified new candidate genetic factors to be screened in the molecular diagnosis of human males with reproductive disorders. Finally, considering the well-known oncogenic nature of these two clusters and the fact that patients with reduced fertility are more prone to testicular cancer, our results might also help to elucidate the molecular mechanisms linking both pathologies.

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Validation of MicroRNA-188-5p Inhibition Power on Tumor Cell Proliferation in Papillary Thyroid Carcinoma

Cell Transplantation ◽

10.1177/0963689720918300 ◽

2020 ◽

Vol 29 ◽

pp. 096368972091830 ◽

Cited By ~ 1

Author(s):

Ping Zhou ◽

Andrew Irving ◽

Huifang Wu ◽

Juan Luo ◽

Johana Aguirre ◽

...

Keyword(s):

Cell Proliferation ◽

Papillary Thyroid Carcinoma ◽

Thyroid Carcinoma ◽

Tumor Cell ◽

Cellular Proliferation ◽

Tumor Cell Proliferation ◽

Papillary Thyroid ◽

Tumor Tissues ◽

Potential Biomarker ◽

And Function

Given the crucial role of microRNAs in the cellular proliferation of various types of cancers, we aimed to analyze the expression and function of a cellular proliferation-associated miR-188-5p in papillary thyroid carcinoma (PTC). Here we demonstrate that miR-188-5p is downregulated in PTC tumor tissues compared with the associated noncancerous tissues. We also validate that the miR-188-5p overexpression suppressed the PTC cancer cell proliferation. In addition, fibroblast growth factor 5 (FGF5) is observed to be downregulated in the PTC tumor tissues compared with the associated noncancerous tissues. Subsequently, FGF5 is identified as the direct functional target of miR-188-5p. Moreover, the silencing of FGF5 was found to inhibit PTC cell proliferation, which is the same pattern as miR-188-5p overexpression. These results suggest that miR-188-5p-associated silencing of FGF5 inhibits tumor cell proliferation in PTC. It also highlights the importance of further evaluating miR-188-5p as a potential biomarker and therapy target in PTC.

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