MiR-491 suppresses migration and invasion via directly targeting TPX2 in breast cancer.

OBJECTIVE: Breast cancer (BC) is one of the primary causes of tumor-related female mortalities. Although in recent years, we have made great progress in the systemic therapy and earlier diagnosis for BC patients, recurrence or distant metastasis remains leading obstacles for the successful therapy of BC. Therefore, a comprehensive understanding of the molecular mechanism underlying the progression may be crucial in developing an effective strategy against BC. The current research aimed to explore the expressions, functions and molecular mechanism of microRNA-491 (miR-491) in BC. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the level of miR-491 expression in 52 pairs of BC tissues and para-cancerous specimens, and the relation between miR-491 level and the clinical features of BC patient prognosis was analyzed. Transwell invasion and migration assays were conducted to determine whether miR-491 had effects on the regulation of BC metastasis. Potential target genes of miR-491 were found out using TargetScan to explore the molecular functions of miR-491 in inhibiting breast cancer cell invasion and migration. To elucidate the mechanism of TPX2 in suppressing cell invasion and migration medicated by miR-491in breast cancer, we further transfected TPX2 siRNAs into MCF-7 cells to delete endogenous TPX2, along with the transfections with miR-491 inhibitor into MCF-7 cell lines. RESULTS: The findings demonstrated that miR-491 expressions were significantly decreased in BC tissues and cells. The miR-491 restoration suppressed the invasion and migration of BC cells. In addition, we identified the targeting protein for Xklp2 (TPX2) as a direct target of miR-491 in BC. The knockdown of TPX2 markedly reversed miR-491-medicated inhibition of cell invasion and migration in BC cell lines. CONCLUSIONS: In short, all the results suggested that miR-491 functioned as a tumor suppressor by targeting TPX2 in BC and the miR-491 restoration may be an effective therapy for the BC treatment in the future.

Combination therapy of orally administered glycyrrhizin and UVB improved active-stage generalized vitiligo.

Glycyrrhizin has been used clinically for several years due to its beneficial effect on immunoglobulin E (IgE)-induced allergic diseases, alopecia areata and psoriasis. In this study, glycyrrhizin, ultraviolet B light (UVB) or a combination of both were used to treat active-stage generalized vitiligo. One hundred and forty-four patients between the ages of 3 and 48 years were divided into three groups: group A received oral compound glycyrrhizin (OCG); group B received UVB applications twice weekly, and group C received OCG+UVB. Follow-ups were performed at 2, 4, and 6 months after the treatment was initiated. The Vitiligo Area Scoring Index (VASI) and the Vitiligo Disease Activity (VIDA) instrument were used to assess the affected body surface, at each follow-up. Results showed that 77.1, 75.0 and 87.5% in groups A, B and C, respectively, presented repigmentation of lesions. Responsiveness to therapy seemed to be associated with lesion location and patient compliance. Adverse events were limited and transient. This study showed that, although the three treatment protocols had positive results, OCG and UVB combination therapy was the most effective and led to improvement in disease stage from active to stable.

Combination therapy of orally administered glycyrrhizin and UVB improved active-stage generalized vitiligo

Glycyrrhizin has been used clinically for several years due to its beneficial effect on immunoglobulin E (IgE)-induced allergic diseases, alopecia areata and psoriasis. In this study, glycyrrhizin, ultraviolet B light (UVB) or a combination of both were used to treat active-stage generalized vitiligo. One hundred and forty-four patients between the ages of 3 and 48 years were divided into three groups: group A received oral compound glycyrrhizin (OCG); group B received UVB applications twice weekly, and group C received OCG+UVB. Follow-ups were performed at 2, 4, and 6 months after the treatment was initiated. The Vitiligo Area Scoring Index (VASI) and the Vitiligo Disease Activity (VIDA) instrument were used to assess the affected body surface, at each follow-up. Results showed that 77.1, 75.0 and 87.5% in groups A, B and C, respectively, presented repigmentation of lesions. Responsiveness to therapy seemed to be associated with lesion location and patient compliance. Adverse events were limited and transient. This study showed that, although the three treatment protocols had positive results, OCG and UVB combination therapy was the most effective and led to improvement in disease stage from active to stable.

Interleukin-33 affects cytokine production by keratinocytes in vitiligo.

BACKGROUND: Vitiligo is an acquired skin pigmentation disorder. Interleukin (IL)-33 is a newly described member of the IL-1 family. AIM: To examine the role of IL-33 and its regulation in vitiligo. METHODS: Expression of IL-33 and its receptor (interleukin 1 receptor-like 1 protein; also known as ST2), in skin biopsies taken from healthy subjects and patients with vitiligo, was examined by immunofluorescence staining and western blotting. IL-33 secretion from primary keratinocytes was measured by ELISA. IL-33-stimulated release of stem cell factor (SCF), basic fibroblast growth factor (bFGF), IL-6 and tumour necrosis factor (TNF)-α from primary keratinocytes was examined by ELISA. RESULTS: IL-33 and ST2 expression was increased in lesional skin, and serum IL-33 was raised in patients with vitiligo. IL-33 expression moved from the nucleus to the cytoplasm of keratinocytes. IL-33 reduced expression of both SCF and bFGF, but increased expression of both IL-6 and TNF-α expression in primary keratinocytes. CONCLUSIONS: IL-33 is secreted by keratinocytes and functions as an alarmin. It may induce melanocyte death by regulating cytokines in the cellular microenvironment.

Expression of aquaporin-4 in human supratentorial meningiomas with peritumoral brain edema and correlation of VEGF with edema formation.

Peritumoral brain edema is a common complication of meningiomas. It is believed that vascular endothelial growth factor (VEGF), as an angiogenic factor, plays a vital role in edema formation. Aquaporin-4 (AQP4) is a small integral membrane protein that regulates water in the normal brain. However, the expression of AQP4 and its relationship to VEGF in edematous meningiomas are not well known. We studied tumor specimens of 59 human supratentorial meningiomas. Western blot analysis was used to detect the expression of AQP4, and double-labeling immunofluorescence histochemical staining was performed to determine the relationship between AQP4 and VEGF. The AQP4 expression was significantly higher in the edema group, in which the protein level was correlated with the extent of edema. Greater VEGF expression was also observed in the edema group, and a relationship between AQP4 and VEGF was found. We conclude that AQP4 is involved in peritumoral brain edema formation in meningiomas and is also closely related to the expression of VEGF.

Promoting of melanocyte adhesion and migration by Malytea Scurfpea fruit in vitro.

The aim of this study was to investigate the effect of Malytea Scurfpea Fruit (MSF) on melanocyte adhesion and migration. Human epidermal melanocytes were treated with MSF and examined for adhesion to bovine serum fibronectin-coated culture dishes. Control and treated cells were also examined for migration into micropore filters coated with the same protein. Compared with control, MSF-treated melanocytes adhered to the dishes more easily and migrated into the filters in a dose-dependent manner. MSF at a dose of more than 200 micro g/ml did not increase melanocyte adhesion and migration accordingly. With the exception of MSF 10 micro g/ml, at every concentration of MSF there were significant differences between treated and untreated melanocytes (p < 0.01) when the adhesion test was studied. Regarding migration, even at a concentration of MSF 10 micro g/ml, obviously increased cell numbers were found compared with MSF untreated melanocytes (p < 0.01). MSF promoted melanocyte adhesion and migration; this could explain, in part, the capacity of MSF to regulate melanocyte function in vitiligo.

Time-dependence and cell-type specificity of synergistic neurotrophin actions on spiral ganglion neurons.

The neurotrophins brain-derived neurotrophin (BDNF) and neurotrophin-3 (NT-3) synergistically enhance survival of spiral ganglion neurons such that simultaneous exposure to both compounds produces a larger response than would be expected from their individual effects. To elucidate the functional role of this neurotrophin interaction, we examined its temporal and cell-type specificity in vitro for both mouse and gerbil spiral ganglion neurons. Synergistic effects were transient; they were maximal within the first two postnatal days and declined during the first postnatal week. Both neurotrophins were, however, still efficacious at increasing cell survival. After postnatal day 10, the effects of coexposure to BDNF and NT-3 were additive rather than synergistic. Synergism declined more rapidly in mouse than gerbil neurons, reflecting the difference in cochlear development for each species. Only neurons without peripherin epitopes, putative type I neurons, showed synergistic survival effects; survival of peripherin-expressing neurons was purely additive. Therefore, during a restricted time period, identical neurotrophin stimuli are capable of preferentially enhancing survival of one class of neurons that compose approximately 95% of the adult spiral ganglion.

Synergistic effects of BDNF and NT-3 on postnatal spiral ganglion neurons.

Neurotrophins have profound effects on the development and maintenance of neurons that compose the VIIIth cranial nerve. In the auditory division of the nerve, brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) have been localized to the sensory epithelium, and their respective high-affinity tyrosine kinase receptors (TrkB and TrkC) are expressed within the neuronal population. By using a culture methodology that allows evaluation of single neurons, we determined that BDNF and neurotrophin-4 (NT-4), which both bind to the TrkB high-affinity receptor, greatly enhanced neuron survival above control cultures. NT-3, which acts via the TrkC high-affinity receptor, also increased survival, but to a lesser extent. By testing a variety of neurotrophin concentrations and combinations, we observed that simultaneous activation of the TrkB and TrkC receptors synergistically promoted neuron survival compared to cultures that contained either neurotrophin alone at the same total concentration. Antibody labeling showed that the high-affinity Trk receptors were localized predominantly to the neurons and not to the surrounding satellite cells; furthermore, TrkB- and TrkC-specific antibodies each labeled 100% of the cultured neurons. These results suggest that synergistic interactions between BDNF and NT-3 may be crucial for spiral ganglion neuron survival during the final stages of development.

Stria vascularis morphogenesis in vitro.

Explants of neonatal murine stria vascularis were maintained in vitro to evaluate the process of morphogenesis in cochlear tissue. Immunohistochemical and electron microscopic studies showed that the relatively undifferentiated cells in culture attained morphological features characteristic of the stria vascularis cell types in vivo (marginal, intermediate and basal cells). The three kinds of cells formed a trilaminated tissue, with the epithelial cells bordering the culture medium, basal-like cells resting on the culture substrate, and the melanocytes layered between. Furthermore, approximately 20% of these cultures displayed a unique alignment of melanocytes which formed elongated bands along the contour of the tissue edge. However, only limited cell extensions were formed between different cell types and interdigitation amongst these processes was abbreviated. Thus, cells from different embryological origins divided, migrated and reestablished appropriate cell-to-cell associations to form a layered tissue similar to the stria vascularis in vivo.

[Genetic diagnosis of pelvic lymph node metastasis in prostate cancer using aspiration biopsy samples].

We have developed a highly sensitive method to detect pelvic lymph node(LN) metastasis using reverse transcriptase-polymerase chain reaction(RT-PCR) with the primers specific for prostate-specific antigen(PSA) gene in combination with the fine needle aspiration biopsy(FNAB). The specimens were obtained from pelvic LN from 15 prostate cancer patients and 15 bladder cancer patients. The aspirated samples (0.05 approximately 0.1 ml) were used for detecting the fragment of PSA mRNA by RT-PCR and Southern blot analysis, and the rest of samples were submitted to conventional cytology. Expression of PSA gene was detected in 9 cases of FNAB samples including all 5 cytologically positive and further more 2 cytologically class III cases, and 2 of 8 cytologically negative cases. RT-PCR of FNAB samples from all cases of bladder cancer were negative for the detection of PSA gene. The sensitivity of PSA gene by RT-PCR was very high and could detect 10 degrees cancer cell. In conclusion, our study suggested that RT-PCR for detection of PSA gene in FNAB samples might become a new diagnostic tool for detection of small foci of prostatic cancer metastasis in LN and combination use of RT-PCR and cytology could greatly contribute to accuracy in diagnosis.

Osteopontin expression detected in adult cochleae and inner ear fluids.

Localization of protein epitopes and mRNA expression showed that there was a wide-spread distribution of osteopontin (OPN) within the membranous labyrinth of the adult mammalian cochleae. Immunoreaction product and mRNA were found within the stria vascularis, VIIIth cranial nerve, spiral ligament and limbus. Only specific cell types within these regions contained abundant OPN mRNA or protein, the main cell type being fibrocytes that populate the spiral limbus and spiral ligament. Epithelial cells that line the luminal surface of the stria vascularis (marginal cells) and neurons that compose the vestibular and auditory ganglia also showed high opn expression. The pattern of anti-OPN staining within membranous labyrinth was comparable to that observed in tissues such as gall bladder, breast and kidney. In those tissues, luminal epithelial cells, corresponding to the marginal cells of the stria vascularis, may be responsible for manufacturing and secreting OPN into the luminal fluids. consistent with those observations, we detected OPN epitopes in cochlear fluids withdrawn from the scalae media and tympani of the cochlea. We found that the protein species in cochlear fluid differed from those present in cerebrospinal fluid (CSF) suggesting that OPN exists in tissue-specific isoforms that may correspond to particular cellular functions.