scholarly journals The Effects of Flavonoid Apigenin on Male Reproductive Health: Inhibition of Spermatogonial Proliferation through Downregulation of Prmt7/Akt3 Pathway

2021 ◽  
Vol 22 (22) ◽  
pp. 12209
Author(s):  
Bingyuan Wang ◽  
Mingrui Zhang ◽  
Jiankang Guo ◽  
Zhiguo Liu ◽  
Rong Zhou ◽  
...  

Apigenin, a common dietary flavonoid abundantly present in a variety of fruits and vegetables, has promising anticancer properties. As an effector of apigenin in myoblasts, protein arginine methyltransferase 7 (Prmt7) is required for male germ cell development. However, whether apigenin may influence male reproductive health through Prmt7 is still unclear. To this end, mouse spermatogonia were treated with different concentrations (2.5 to 50 μM) of apigenin for 48 h, which showed that apigenin could cause reduced cell proliferation in conjunction with longer S phase and G2/M phase (with concentrations of 10 and 20 μM, respectively), and increased apoptosis of spermatogonia (with concentration of 20 μM). Reduced Prmt7 expression was found in 20 μM apigenin-treated spermatogonia. Moreover, siRNA-induced Prmt7 knockdown exhibited similar influence on spermatogonia as that of apigenin treatment. In mechanistic terms, transcriptome analysis revealed 287 differentially expressed genes between Prmt7-downregulated and control spermatogonia. Furthermore, rescue experiments suggested that the effects of apigenin on spermatogonia might be mediated through the Prmt7/Akt3 pathway. Overall, our study supports that apigenin can interfere with mouse spermatogonial proliferation by way of the downregulated Prmt7/Akt3 pathway, which demonstrates that the concentration should be taken into account in future applications of apigenin for cancer therapy of men.

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 5377-5377 ◽  
Author(s):  
Mari Cleia Martins Rodrigues Ferreira ◽  
Renata Kikuchi Foltran ◽  
Rodrigo Santucci ◽  
Luis Alberto de Padua Covas Lage ◽  
Debora Levy ◽  
...  

Abstract Introduction: Adult T-cell leukemia/lymphoma (ATLL) is an aggressive and incurable disease caused by human T-lymphotropic virus 1 (HTLV-1) that infects CD4+ and CD8+ lymphocytes, but most commonly the malignant cell present a CD4+ phenotype. However, clonal expansion and cell cycle abnormalities have been demonstrated in CD4+ and in CD8+ lymphocytes of HTLV-1 carriers. Objectives: This study compared DNA content and G0/G1, G2/M and “S”-phases of CD4+ and CD8+ lymphocytes among asymptomatic HTLV-1 carriers, ATLL and health subjects. Methods: Werestudied 38 HTLV-1 carriers, 20 ATLL and 35 health subjects pared by sex and age at the Hematology Department of the Faculty of Medicine, University of São Paulo. Peripheral blood mononuclear cells (PBMCs) were isolated on Ficoll-Paque® and lymphocytes subtypes were obtained by positive selection in a magnetic column. Cell-cycle distribution and DNA index (DI) was assessed by flow cytometry after propidium iodide staining. Results: In ATLL, themedian age was 53.5 years (24 to 72) and 50% were female, in HTLV-1 carriers was 55.5 years (33 to 80) with 63.2% of female and in control group was 50 years (24 to 80) with 54.3% of female. In the CD4+ lymphocyte a % of cells in G0/G1 (98.32%) in HTLV-1 carriers was higher than in control group (97.14%) (p=0.041) and in ATLL (97.25%) (p=0.023). S-phase was not statistically different in asymptomatic carriers (0.34%) and control group (0.63%) (p=0.073), but was higher in ATLL (1.80%) than in asymptomatic carriers (0.34%) (p<0.001) and in control group (0.63%) (p=0.02). G2/M-phase was not significantly different among all groups (p=0.960) (Table 1). The CD4+ lymphocytes were aneuploidy in 39.5% (18.4% DI > 1.05 and 21.5% < 0.95) of asymptomatic carriers and in 26.7% (20% > 1.05 and 6.7% < 0.95) of ATLL patients (p=0.557). All control groups were diploid. Table 1.Comparison of the cell cycle by flow cytometry of T lymphocytes CD4+CD4+ cellsAsymptomatic carriersATLLControl groupp-ValueG0/G1mean(dp)97.78 (2.182)95.69 (3.557)96.55 (2.964)0.0351º; median;3ºq97.03;98.32;99.6491.40;97.25;98.3295.01;97.14;98.64G2/Mmean(dp)1.55(1.848)1.91(2.798)2.03(2.902)0.961º; median;3ºq0.00;0.88;2.670.12;0.99;1.990.00;0.56;2.97S-phasemean(dp)0.68(1.207)2.80(3.372)1.43(1.780)0.0031º; median;3ºq0.00;0.34;0.650.65;1.80;3.510.04;0.63;2.55 In CD8+ there was no found significantly difference in whole groups for G0/G1-phase (p=0.138) and G2/M-phase (p=0.374). ATLL presented higher S-phase (median 1.54%) than asymptomatic carriers (median 0.45%) (p=0.003) and control group. S-phase in asymptomatic carriers was not significantly different in comparison to control group (p=0.712). CD8+ were aneuploidy in 23.7% (5.3% DI > 1.05 and 18.4% < 0.95) of asymptomatic carriers and in 21% (10.5% > 1.05 and 10.5% < 0.95) of ATLL (p=0.603). In ATLL the median of DI was 1.01 (1.0; 1.05) in CD4+ and higher than in CD8+ median 0.99 (0.98; 1.0) (p=0.007). Aneuploidia was seen in 47.7% of ATLL, 26,7% (20% DI > 1.05 and 6,7% < 0.95) in CD4+ and 21,0% in CD8+ (10,5% > 1.05 and10,5% < 0.95) (p=0.625). Figure 1: Dna index of CD4+ and CD8+. Aneuploidia was found in HTLV I carriers in both CD4+ and CD8+. Figure 1:. Dna index of CD4+ and CD8+. Aneuploidia was found in HTLV I carriers in both CD4+ and CD8+. Figure 2. Comparison of DI between CD4+ and CD8+ of asymptomatic carriers and ATLL Figure 2. Comparison of DI between CD4+ and CD8+ of asymptomatic carriers and ATLL Figure 3 Figure 3. Conclusion: We demonstrated for the first time “in vivo” that asymptomatic HTLV-1 carriers display cell cycle arrest in G0/G1-phase in CD4+ lymphocytes and high rate of aneuploidia in both CD4+ and CD8+. ATLL showed high rate of hiperdiploidia in CD4+ and hipodiploidia in CD8+ and high rate of S-phase in CD4+. Genetic instability and proliferative disturbs are a hallmark not only in ATLL but also in HTLV-1 carriers and in both CD4+ and CD8+ lymphocytes. Disclosures No relevant conflicts of interest to declare.


2021 ◽  
Author(s):  
Daniel E. Nassau ◽  
Jordan C. Best ◽  
Eliyahu Kresch ◽  
Daniel C Gonzalez ◽  
Kajal Khodamoradi ◽  
...  

2021 ◽  
Author(s):  
P Stamatiadis ◽  
A Boel ◽  
G Cosemans ◽  
M Popovic ◽  
B Bekaert ◽  
...  

Abstract STUDY QUESTION What is the role of POU class 5 homeobox 1 (POU5F1) in human preimplantation development and how does it compare with the mouse model? SUMMARY ANSWER POU5F1 is required for successful development of mouse and human embryos to the blastocyst stage as knockout embryos exhibited a significantly lower blastocyst formation rate, accompanied by lack of inner cell mass (ICM) formation. WHAT IS KNOWN ALREADY Clustered regularly interspaced short palindromic repeats—CRISPR associated genes (CRISPR-Cas9) has previously been used to examine the role of POU5F1 during human preimplantation development. The reported POU5F1-targeted blastocysts always retained POU5F1 expression in at least one cell, because of incomplete CRISPR-Cas9 editing. The question remains of whether the inability to obtain fully edited POU5F1-targeted blastocysts in human results from incomplete editing or the actual inability of these embryos to reach the blastocyst stage. STUDY DESIGN, SIZE, DURATION The efficiency of CRISPR-Cas9 to induce targeted gene mutations was first optimized in the mouse model. Two CRISPR-Cas9 delivery methods were compared in the B6D2F1 strain: S-phase injection (zygote stage) (n = 135) versus metaphase II-phase (M-phase) injection (oocyte stage) (n = 23). Four control groups were included: non-injected media-control zygotes (n = 43)/oocytes (n = 48); sham-injected zygotes (n = 45)/oocytes (n = 47); Cas9-protein injected zygotes (n = 23); and Cas9 protein and scrambled guide RNA (gRNA)-injected zygotes (n = 27). Immunofluorescence analysis was performed in Pou5f1-targeted zygotes (n = 37), media control zygotes (n = 19), and sham-injected zygotes (n = 15). To assess the capacity of Pou5f1-null embryos to develop further in vitro, additional groups of Pou5f1-targeted zygotes (n = 29) and media control zygotes (n = 30) were cultured to postimplantation stages (8.5 dpf). Aiming to identify differences in developmental capacity of Pou5f1-null embryos attributed to strain variation, zygotes from a second mouse strain—B6CBA (n = 52) were targeted. Overall, the optimized methodology was applied in human oocytes following IVM (metaphase II stage) (n = 101). The control group consisted of intracytoplasmically sperm injected (ICSI) IVM oocytes (n = 33). Immunofluorescence analysis was performed in human CRISPR-injected (n = 10) and media control (n = 9) human embryos. PARTICIPANTS/MATERIALS, SETTING, METHODS A gRNA-Cas9 protein mixture targeting exon 2 of Pou5f1/POU5F1 was microinjected in mouse oocytes/zygotes or human IVM oocytes. Reconstructed embryos were cultured for 4 days (mouse) or 6.5 days (human) in sequential culture media. An additional group of mouse-targeted zygotes was cultured to postimplantation stages. Embryonic development was assessed daily, with detailed scoring at late blastocyst stage. Genomic editing was assessed by immunofluorescence analysis and next-generation sequencing. MAIN RESULTS AND THE ROLE OF CHANCE Genomic analysis in mouse revealed very high editing efficiencies with 95% of the S-Phase and 100% of the M-Phase embryos containing genetic modifications, of which 89.47% in the S-Phase and 84.21% in the M-Phase group were fully edited. The developmental capacity was significantly compromised as only 46.88% embryos in the S-Phase and 19.05% in the M-Phase group reached the blastocyst stage, compared to 86.36% in control M-Phase and 90.24% in control S-Phase groups, respectively. Immunofluorescence analysis confirmed the loss of Pou5f1 expression and downregulation of the primitive marker SRY-Box transcription factor (Sox17). Our experiments confirmed the requirement of Pou5f1 expression for blastocyst development in the second B6CBA strain. Altogether, our data obtained in mouse reveal that Pou5f1 expression is essential for development to the blastocyst stage. M-Phase injection in human IVM oocytes (n = 101) similarly resulted in 88.37% of the POU5F1-targeted embryos being successfully edited. The developmental capacity of generated embryos was compromised from the eight-cell stage onwards. Only 4.55% of the microinjected embryos reached the late blastocyst stage and the embryos exhibited complete absence of ICM and an irregular trophectoderm cell layer. Loss of POU5F1 expression resulted in absence of SOX17 expression, as in mouse. Interestingly, genetic mosaicism was eliminated in a subset of targeted human embryos (9 out of 38), three of which developed into blastocysts. LIMITATIONS, REASONS FOR CAUTION One of the major hurdles of CRISPR-Cas9 germline genome editing is the occurrence of mosaicism, which may complicate phenotypic analysis and interpretation of developmental behavior of the injected embryos. Furthermore, in this study, spare IVM human oocytes were used, which may not recapitulate the developmental behavior of in vivo matured oocytes. WIDER IMPLICATIONS OF THE FINDINGS Comparison of developmental competency following CRISPR-Cas-mediated gene targeting in mouse and human may be influenced by the selected mouse strain. Gene targeting by CRISPR-Cas9 is subject to variable targeting efficiencies. Therefore, striving to reduce mosaicism can provide novel molecular insights into mouse and human embryogenesis. STUDY FUNDING/COMPETING INTEREST(S) The research was funded by the Ghent University Hospital and Ghent University and supported by the FWO-Vlaanderen (Flemish fund for scientific research, Grant no. G051516N), and Hercules funding (FWO.HMZ.2016.00.02.01). The authors declare no competing interests. TRIAL REGISTRATION NUMBER N/A.


Andrology ◽  
2019 ◽  
Vol 7 (6) ◽  
pp. 761-761
Author(s):  
D. T. Carrell ◽  
M. Simoni ◽  
C. Krausz ◽  
G. L. Gerton

2000 ◽  
Vol 74 (19) ◽  
pp. 9152-9166 ◽  
Author(s):  
Grace Y. Lin ◽  
Robert A. Lamb

ABSTRACT Infection of cells by many viruses affects the cell division cycle of the host cell to favor viral replication. We examined the ability of the paramyxovirus simian parainfluenza virus 5 (SV5) to affect cell cycle progression, and we found that SV5 slows the rate of proliferation of HeLa T4 cells. The SV5-infected cells had a delayed transition from G1 to S phase and prolonged progression through S phase, and some of the infected cells were arrested in G2 or M phase. The levels of p53 and p21CIP1were not increased in SV5-infected cells compared to mock-infected cells, suggesting that the changes in the cell cycle occur through a p53-independent mechanism. However, the phosphorylation of the retinoblastoma protein (pRB) was delayed and prolonged in SV5-infected cells. The changes in the cell cycle were also observed in cells expressing the SV5 V protein but not in the cells expressing the SV5 P protein or the V protein lacking its unique C terminus (VΔC). The unique C terminus of the V protein of SV5 was shown previously to interact with DDB1, which is the 127-kDa subunit of the multifunctional damage-specific DNA-binding protein (DDB) heterodimer. The coexpression of DDB1 with V can partially restore the changes in the cell cycle caused by expression of the V protein.


2016 ◽  
Vol 42 (4) ◽  
pp. 454-462 ◽  
Author(s):  
L. V. Perelomov ◽  
I. V. Perelomova ◽  
U. L. Venevtseva

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