scholarly journals Changes in the expression and functional activities of Myosin II isoforms in human hyperplastic prostate

2021 ◽  
Author(s):  
Weixiang He ◽  
Xiao Wang ◽  
Daxing Zhan ◽  
Mingzhou Li ◽  
Qian Wang ◽  
...  
Keyword(s):  
Myosin Ii ◽  
Human Prostate ◽  
Common Disease ◽  
Organ Bath ◽  
Prostate Cell ◽  
Functional Activities ◽  
Prostate Cells ◽  
Induced Apoptosis ◽  
Muscle Myosin

Benign prostatic hyperplasia (BPH) is a common disease among aging males with the etiology remaining unclear. We recently found myosin II was abundantly expressed in rat and cultured human prostate cells with permissive roles in the dynamic and static components. This study aimed to explore the expression and functional activities of myosin II isoforms including smooth muscle myosin II (SMM II) and non-muscle myosin (NMM II) in the hyperplastic prostate. Human prostate cell lines and tissues from normal human and BPH patients were used. H&E, Masson’s trichrome, immunohistochemical staining, in vitro organ bath, RT-PCR and Western-blotting were performed. We further created cell models with NMM II isoforms silenced and proliferation, cycle, and apoptosis of prostate cells were determined by CCK-8 assay and flow cytometry. Hyperplastic prostate SM expressed more SM1 and LC17b isoforms compared to their alternatively spliced counterparts, favoring a slower more tonic-type contraction and greater force generation. For BPH group, blebbistatin (BLEB, a selective myosin II inhibitor), exhibited a stronger effect on relaxing phenylephrine (PE) pre-contracted prostate strips and inhibiting PE induced contraction. Additionally, NMMHC-A and NMMHC-B were upregulated in hyperplastic prostate with no change in NMMHC-C. Knockdown of NMMHC-A or NMMHC-B inhibited prostate cell proliferation and induced apoptosis, with no changes in cell cycle. Our novel data demonstrates that expression and functional activities of myosin II isoforms are altered in human hyperplastic prostate, suggesting a new pathological mechanism for BPH. Thus, the myosin II system may provide potential new therapeutic targets for BPH/lower urinary tract symptoms (LUTS).

scholarly journals Smooth muscle myosin expression, isoform composition, and functional activities in rat corpus cavernosum altered by the streptozotocin-induced type 1 diabetes

2012 ◽  
Vol 302 (1) ◽  
pp. E32-E42 ◽  
Author(s):  
Xinhua Zhang ◽  
Nirmala D. Kanika ◽  
Arnold Melman ◽  
Michael E. DiSanto
Keyword(s):  
Smooth Muscle ◽  
Alternative Splicing ◽  
Corpus Cavernosum ◽  
Smooth Muscle Myosin ◽  
Organ Bath ◽  
Rt Pcr ◽  
Functional Activities ◽  
Muscle Myosin

Diabetes mellitus (DM) is a quite common chronic disease, and the prevalence of erectile dysfunction (ED) is three times higher in this large population. Although diabetes-related ED has been studied extensively, the actin-myosin contractile apparatus was not examined. The mRNAs encoding smooth muscle myosin (SMM) heavy chains (MHC) and essential light chains (LC17) exist as several different alternatively spliced isoforms with distinct contractile properties. Recently, we provided novel data that blebbistatin (BLEB), a specific myosin II inhibitor, potently relaxed corpus cavernosum smooth muscle (CCSM). In this study, we examine whether diabetes alters SMM expression, alternative splicing, and/or functional activities, including sensitivity to BLEB. By using streptozotocin (STZ)-induced 2-mo diabetic rats, functional activities were tested in vivo by intracavernous pressure (ICP) recording during cavernous nerve stimulation and in vitro via organ bath contractility studies. SMM isoform composition was analyzed by competitive RT-PCR and total SMM, myocardin, and embryonic SMM (SMemb) expression by real-time RT-PCR. Results revealed that the blood glucose level of STZ rats was 407.0 vs. 129.5 mg/dl (control). STZ rats exhibited ED confirmed by significantly increased CCSM contractile response to phenylephrine and decreased ICP response. For STZ rats, SM-B, LC17a and SM2 isoforms, total SMM, and myocardin expression increased, whereas SM-A, LC17b, and SM1 isoforms were decreased, with SMemb unchanged. BLEB was significantly more effective in relaxing STZ CCSM both in vitro and in vivo. Thus we demonstrated a novel diabetes-specific effect on alternative splicing of the SMM heavy chain and essential light chain genes to a SMM isoform composition favoring a heightened contractility and ED. A switch to a more contractile phenotype was supported further by total SMM expression increase. Moreover, the change in CCSM phenotype was associated with an increased sensitivity to BLEB, which may serve as a novel pharmacotherapy for ED.

scholarly journals Dia- and Rok-dependent enrichment of capping proteins in a cortical region

2021 ◽  
Author(s):  
Anja Schmidt ◽  
Long Li ◽  
Zhiyi Lv ◽  
Shuling Yan ◽  
Jörg Großhans
Keyword(s):  
Myosin Ii ◽  
Rho Kinase ◽  
Cortical Region ◽  
Protein Enrichment ◽  
Cortical Actin ◽  
Capping Protein ◽  
Capping Proteins ◽  
Muscle Myosin ◽  
Drosophila Embryos

Rho signaling with its major targets the formin Dia, Rho kinase (Rok) and non-muscle myosin II control turnover, amount and contractility of actomyosin. Much less investigated has been a potential function for the distribution of F-actin plus and minus ends. In syncytial Drosophila embryos Rho1 signaling is high between actin caps, i. e. the cortical intercap region. Capping protein binds to free plus ends of F-actin to prevent elongation of the filament. Capping protein has served as a marker to visualize the distribution of F-actin plus ends in cells and in vitro. Here, we probed the distribution of plus ends with capping protein in syncytial Drosophila embryos. We found that Capping proteins are specifically enriched in the intercap region similar to Dia and MyoII but distinct from overall F-actin. The intercap enrichment of Capping protein was impaired in dia mutants and embryos, in which Rok and MyoII activation was inhibited. Our observations reveal that Dia and Rok/MyoII control Capping protein enrichment and support a model that Dia and Rok/MyoII control the organization of cortical actin cytoskeleton downstream of Rho1 signaling.

scholarly journals Androgen receptor localisation and turnover in human prostate epithelium treated with the antiandrogen, casodex

2000 ◽  
Vol 24 (3) ◽  
pp. 339-351 ◽  
Author(s):  
AS Waller ◽  
RM Sharrard ◽  
P Berthon ◽  
NJ Maitland
Keyword(s):  
Androgen Receptor ◽  
In Vitro Models ◽  
Human Prostate ◽  
Intracellular Targeting ◽  
Natural Ligand ◽  
Prostate Cells ◽  
Established Cell ◽  
Prostatic Epithelium ◽  
Prostate Epithelium

In vitro models of normal and malignant human prostate are currently limited to a few well established cell lines that, with a single exception (LNCaP), fail to express the androgen receptor (AR) - a common characteristic of prostatic epithelium grown in culture. To investigate the molecular mechanism of action of the non-steroidal antiandrogen Casodex (bicalutamide) against wild-type AR, we have established a transient AR expression model in non-tumorigenic prostate cells of both epithelial and mesenchymal origin. In this model, both dihydrotestosterone and Casodex can effectively transport the AR protein into the nucleus of prostate cells. Whereas the natural ligand, dihydrotestosterone, stabilises the receptor, the AR is rapidly degraded at a nuclear location when the transfected cells are treated with Casodex. In contrast, whereas the mutant AR in the LNCaP line is also degraded on Casodex treatment over the same time period, its intracellular targeting is defective.

scholarly journals Testosterone regulates smooth muscle contractile pathways in the rat prostate: emphasis on PDE5 signaling

2012 ◽  
Vol 302 (2) ◽  
pp. E243-E253 ◽  
Author(s):  
Xinhua Zhang ◽  
Ning Zang ◽  
Yu Wei ◽  
Jin Yin ◽  
Ruobing Teng ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Male Rats ◽  
Nitric Oxide Donor ◽  
Organ Bath ◽  
Functional Activities ◽  
Urinary Tract Symptoms ◽  
Mrna And Protein Expression ◽  
Rat Prostate ◽  
Prostate Stroma

Testosterone (T) plays a permissive role in the development of benign prostatic hyperplasia (BPH), and phosphodiesterase 5 inhibitors (PDE5is) have been found to be effective for BPH and lower urinary tract symptoms (LUTS) in clinical trials. This study investigated the effect of T on smooth muscle (SM) contractile and regulatory signaling pathways, including PDE5 expression and functional activity in prostate in male rats (sham-operated, surgically castrated, and castrated with T supplementation). In vitro organ bath studies, real-time RT-PCR, Western blot analysis, and immunohistochemistry were performed. Castration heavily attenuated contractility, including sensitivity to phenylephrine with SM myosin immunostaining revealing a disrupted SM cell arrangement in the stroma. PDE5 was immunolocalized exclusively in the prostate stroma, and orchiectomy signficantly reduced PDE5 immunopositivity, mRNA, and protein expression, along with nNOS and ROKβ mRNA, whereas it increased eNOS plus α1a and α1b adrenoreceptor expression in castrated animals. The PDE5i zaprinast significantly increased prostate strip relaxation to the nitric oxide donor sodium nitroprusside (SNP) in control but not castrated rats. But SNP alone was more effective on castrated rats, comparable with sham treated with SNP plus zaprinast. T supplementation prevented or restored all above changes, including SNP and zaprinast in vitro responsiveness. In conclusion, our data show that T positively regulates PDE5 expression and functional activities in prostate, and T ablation not only suppresses prostate size but also reduces prostatic SM contractility, with several potential SM contraction/relaxation pathways implicated. Zaprinast findings strongly suggest a major role for PDE5/cGMP in this signaling cascade. PDE5 inhibition may represent a novel mechanism for treatment of BPH.

scholarly journals UNC-45A Weakens and Breaks MT Lattice Independent of its Effect on Non-Muscle Myosin II

2020 ◽  
Author(s):  
Juri Habicht ◽  
Ashley Mooneyham ◽  
Asumi Hoshino ◽  
Mihir Shetty ◽  
Xiaonan Zhang ◽  
...  
Keyword(s):  
Neurodegenerative Diseases ◽  
Myosin Ii ◽  
Human Diseases ◽  
Therapeutic Implications ◽  
Associated Functions ◽  
Muscle Myosin ◽  
First Time ◽  
In Cells

AbstractIn invertebrates, UNC-45 regulates myosin stability and functions. Vertebrates have two distinct isoforms of the protein: UNC-45B, expressed in muscle cells only and UNC-45A, expressed in all cells and implicated in regulating both Non-Muscle Myosin II (NMII)- and microtubule (MT)-associated functions. Here we show for the first time that: a) in vitro UNC-45A binds to the MT lattice and weakens its integrity leading to MT bending, breakage and depolymerization, b) in cells, UNC-45A overexpression causes loss of MT mass and increase in MT breakages, c) both in vitro and in cells, UNC-45A destabilizes MTs independent of its NMII C-terminal binding domain and destabilization occurs even in presence of the NMII inhibitor blebbistatin. These findings are consistent with a not mutually exclusive but rather dual role of UNC-45A in regulating NMII activity and MT stability.Because many human diseases, from cancer to neurodegenerative diseases, are caused by or associated with deregulation of MT stability our findings have profound implications in both, the biology of MTs as well as the biology of human diseases and possible therapeutic implications for their treatment.

307. The effect of oxytocin on cell growth and steroid production in normal human prostate cells in vitro

2005 ◽  
Vol 17 (9) ◽  
pp. 131
Author(s):  
K. J. Hogarth ◽  
K. King ◽  
H. D. Nicholson
Keyword(s):  
Cell Growth ◽  
Epithelial Cells ◽  
Stromal Cell ◽  
Human Prostate ◽  
Steroid Production ◽  
Cell Numbers ◽  
Prostate Cells ◽  
Normal Human ◽  
Oxytocin Antagonist

Oxytocin (OT) is present in reproductive tissues of male mammals including human prostate tissue. OT increases prostatic muscle tone and prostatic growth. OT is increased in benign prostatic hyperplasia (BPH), an androgen dependent condition that develops with age. Dihydrotestosterone (DHT) is the active hormone in the prostate and is converted from testosterone by the enzyme 5 a reductase. Conversion has been shown to be augmented in the presence of OT. The aim of this study was to investigate the effect of oxytocin on cell growth and steroid production in cultured normal human prostate cells. Normal human prostate stromal and epithelial cells (Clonetics) were cultured with OT, oxytocin antagonist (OTA) or oxytocin/oxytocin antagonist combination (10 ng/mL, 1 ng/mL or 0.1 ng/mL) in media containing 10 nmol of testosterone. Media was changed daily over the 5 day growth period and frozen. Cell proliferation assay was performed at harvest on day 5 to ascertain cell numbers. Media from days 1, 3 and 5 were extracted and radioimmunoassayed for testosterone and DHT. OT increased stromal cell number in a dose dependent manner (P < 0.001). Treatment with OT or OTA had no significant effect on epithelial cell numbers. In stromal cell media from Day 1, DHT concentrations were higher in cells treated with OT than control cells (P < 0.05). By Day 5 the concentration of DHT was low in all treatment groups except OT (10 ng/mL). No effect of OT or OTA was seen on DHT concentrations of media from epithelial cells. OT may increase cell growth in prostate stromal cells but not epithelial cells grown in vitro. This effect may be related to the conversion of testosterone to DHT and DHT to its metabolites. These results demonstrate that OT may play a role in the regulation of cell growth, steroid production and steroid metabolism in the human prostate.

scholarly journals Phosphorylation of myosin-II regulatory light chain by cyclin-p34cdc2: a mechanism for the timing of cytokinesis.

1992 ◽  
Vol 118 (3) ◽  
pp. 595-605 ◽  
Author(s):  
L L Satterwhite ◽  
M J Lohka ◽  
K L Wilson ◽  
T Y Scherson ◽  
L J Cisek ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Chromosome Segregation ◽  
Light Chain ◽  
Kinase Activity ◽  
Myosin Ii ◽  
Regulatory Light Chain ◽  
Nonmuscle Myosin ◽  
Xenopus Egg ◽  
Muscle Myosin

To understand how cytokinesis is regulated during mitosis, we tested cyclin-p34cdc2 for myosin-II kinase activity, and investigated the mitotic-specific phosphorylation of myosin-II in lysates of Xenopus eggs. Purified cyclin-p34cdc2 phosphorylated the regulatory light chain of cytoplasmic and smooth muscle myosin-II in vitro on serine-1 or serine-2 and threonine-9, sites known to inhibit the actin-activated myosin ATPase activity of smooth muscle and nonmuscle myosin (Nishikawa, M., J. R. Sellers, R. S. Adelstein, and H. Hidaka. 1984. J. Biol. Chem. 259:8808-8814; Bengur, A. R., A. E. Robinson, E. Appella, and J. R. Sellers. 1987. J. Biol. Chem. 262:7613-7617; Ikebe, M., and S. Reardon. 1990. Biochemistry. 29:2713-2720). Serine-1 or -2 of the regulatory light chain of Xenopus cytoplasmic myosin-II was also phosphorylated in Xenopus egg lysates stabilized in metaphase, but not in interphase. Inhibition of myosin-II by cyclin-p34cdc2 during prophase and metaphase could delay cytokinesis until chromosome segregation is initiated and thus determine the timing of cytokinesis relative to earlier events in mitosis.

Effect of a standardized extract of red orange juice on proliferation of human prostate cells in vitro

Fitoterapia ◽  
2006 ◽  
Vol 77 (3) ◽  
pp. 151-155 ◽  
Author(s):  
Federica Vitali ◽  
Claudia Pennisi ◽  
Antonio Tomaino ◽  
Francesco Bonina ◽  
Anna De Pasquale ◽  
...  
Keyword(s):  
Orange Juice ◽  
Human Prostate ◽  
Prostate Cells
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