MiR-15b facilitates breast cancer progression via repressing tumor suppressor PAQR3.

OBJECTIVE: Recently, breast cancer (BC) has become a common tumor that threatens the physical and mental health of women. Microribonucleic acids (miRNAs) have been chosen as a study object because of their roles in various cancers, including BC. Here, we mainly study the role of miR-15b in BC progression and its underlying mechanism. PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine the level of MiR-15b expression in 60 pairs of BC tissues and para-cancerous specimens, and the relationship between MiR-15b level and clinical features of BC patient prognosis was analyzed. MiR-15b and PAQR3 level in BC tissues and cells was tested by Western blot. RESULTS: The results showed that miR-15b expression was higher and PAQR3 level was lower in BC. The identification of PAQR3 as a target of miR-15b in BC was carried out by Luciferase reporter assay and the results stated that the Luciferase activity was reduced by miR-15b mimic, indicating PAQR3 being a target of miR-15b in BC. Transwell assay was used for examining BC cell migration and invasion and found that miR-15b could promote BC cell migration and invasion, while the effect of PAQR3 was inhibition. Furthermore, PAQR3 could reverse the promotion effect of miR-15b on BC cell migratory and invasive ability. In addition, miR-15b expression was negatively correlated with PAQR3 performed by regression analysis. CONCLUSIONS: Our data stated that miR-15b could facilitate BC progression via repressing tumor suppressor PAQR3, indicating that miR-15b/PAQR3 axis provided a therapeutic target for treating BC.

MicroRNA-486 inhibits cell proliferation, invasion and migration via down-regulating the TENM1 expressions and affecting ERK and Akt signaling pathways and epithelial-to-mesenchymal transition in papillary thyroid carcinoma.

OBJECTIVE: Papillary thyroid carcinoma (PTC) is one of the general thyroid malignancies. Recently, microRNAs (miRNAs) have identified as pivotal gene regulators in PTC tumorigenesis. The aim of this study was to investigate the role of miR-486 in PTC and its underlying mechanism. PATIENTS AND METHODS: Fifty-six pairs of PTC tissue and matched normal tissue samples were collected from PTC patients who underwent surgery at our hospital from March 2015 to September 2017. Human thyroid epithelial cell line Nthy-ori3-1and PTC cell lines (BCPAP, K1, HTH83, and TPC-1) were cultured. The mRNA and protein expression level were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Additionally, the proliferation and migration abilities were checked by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) method and transwell assay, respectively. Furthermore, dual-luciferase reporter assay was performed to confirm the combination of miR-486 and TENM1. Xenograft Model experiments were performed to assess the effects of miR-486 on tumor growth in vivo. RESULTS: MiR-486 expression was significantly reduced in PTC, which was associated with the poorer clinicopathologic characteristics and overall survival (OS) of PTC patients. Moreover, miR-486 restoration in PTC cells was confirmed to markedly inhibit proliferation, invasion, and migration via the regulation of extracellular-signal-regulated kinase (ERK) and protein kinase B (Akt) signaling pathways and epithelial-mesenchymal transition (EMT). In the meantime, teneurin transmembrane protein 1 (TENM1) was identified as a direct functional target for miR-486 in PTC cells on the basis of bioinformatic analysis and luciferase reporter assays. Additionally, we also verified that miR-486 restoration could prominently repress the PTC growth in vivo. CONCLUSIONS: MiR-486 exerted anti-tumor functions in PTC progression and served as promising biomarkers for the PTC treatment.

LncRNA SNHG20 promotes the development of laryngeal squamous cell carcinoma by regulating miR-140.

OBJECTIVE: This study aims to investigate the expression level of long non-coding RNA (lncRNA) SNHG20 in laryngeal squamous cell carcinoma (LSCC), and to explore further whether it can promote the development of LSCC by regulating microRNA-140 (miR-140). PATIENTS AND METHODS: Expression levels of SNHG20 in 56 pairs of LSCC tissues and adjacent normal tissues were measured by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The relationship between SNHG20 expression with pathological parameters and the prognosis of LSCC was analyzed. Besides, the SNHG20 expression in LSCC cells was also analyzed by qRT-PCR. The SNHG20 knockdown and overexpression model were constructed by lentivirus transfection in AMC-HN-8 and Hep-2 cells. Cell counting kit-8 (CCK-8) and 5-Ethynyl-2'-deoxyuridine (EdU) assay were used to analyze the effect of SNHG20 on the biological function of LSCC cells. Finally, the dual-luciferase reporter gene assay was performed to explore the potentials of SNHG20 and miR-140 in LSCC. RESULTS: The SNHG20 expression in LSCC tissues or cells remarkably increased than controls, and the difference was statistically significant. The LSCC patients with the high expression level of SNHG20 were more likely to develop advanced tumor compared with patients with low expression of SNHG20. Moreover, the LSCC patients with the high expression level of SNHG20 had a shorter overall survival than those with low level. The cell proliferation ability significantly decreased in the SNHG20 knockdown group, while notably increased in SNHG20 overexpression group. MiR-140 was negatively correlated with SNHG20 in LSCC tissues and cells. Dual-luciferase reporter gene assay showed that SNHG20 could be targeted by miR-140 through a certain binding site. The cell rescue experiment also indicated that there was a mutual regulation between SNHG20 and miR-140, which could together affect the malignant progression of LSCC. CONCLUSIONS: We showed that the expression levels of SNHG20 in LSCC tissues or cell lines significantly increased and was associated with advanced tumor staging and undesirable prognosis of LSCC. In addition, SNHG20 could promote the malignant progression of LSCC.

[The current status and applications of implantable bone-conduction devices].

Implantable bone-conduction devices are characterized by the fact that the vibration is transmitted through bone conduction. The technology and surgical techniques in the application of implantable bone-conduction devices have developed considerably in recent years, experiencing a transformation from percutaneous to transcutaneous implantation. This article reviewed current developments in the types, surgical indications, and complications, as well as compared between the various bone-conduction devices to provid references for clinical application.

Surgery alone versus post-operative radiotherapy for sinonasal malignant melanoma: a meta-analysis.

OBJECTIVE: Sinonasal malignant melanoma is a relatively rare malignancy with poor prognosis, and effective treatments remain elusive. This analysis aimed to explore whether post-operative radiotherapy conferred any survival advantages in patients with this disease when compared with surgery alone. METHODS: Published studies were identified by searching four electronic databases. The endpoints evaluated were: rates of overall survival, disease-free survival and local control. RESULTS: Twenty-eight studies including 1392 patients were identified. The results indicated that post-operative radiotherapy led to a significantly better three-year overall survival rate (p = 0.02), and suggested a borderline significant benefit for five-year overall survival (p = 0.05), when compared with surgery alone. However, no statistical advantage was found for disease-free survival, local control or one-year overall survival. CONCLUSION: This meta-analysis indicated that adjuvant radiotherapy prolonged survival, but showed no benefit for disease-free survival or local control.

[Effects of rabbit adipose-derived mesenchymal stem cells on the healing of skin deep partial-thickness scald wound of rabbit].

OBJECTIVE: To investigate the effects of local injection of rabbit adipose-derived mesenchymal stem cells (ADSCs) on the healing of skin deep partial-thickness scald wound of rabbit. METHODS: ADSCs were isolated from adipose tissue of one New Zealand rabbit and then sub-cultured. ADSCs of the third passage were used in the following experiments. Twenty-four rabbits were divided into ADSCs group (n=12) and control group (n=12) according to the random number table, and one deep partial-thickness scald wound with diameter of 5 cm on the two sides of the back near the buttocks was made. From post injury day (PID) 2, 2 mL suspension of EdU-labeled ADSCs with the number of 5×10(5) per mL was subcutaneously injected in wounds of rabbits in ADSCs group, while the rabbits in control group were given 2 mL serum-free DMEM until the wounds were healed. Wound healing processes of rabbits in two groups were observed every day, and the healing time was recorded. On PID 7, 14, 21, and 28, areas of wound of three rabbits in two groups were measured and the healing rates were calculated, respectively. The healed wound tissue was harvested to observe the morphology by HE staining, and the expression of collagen fiber was observed by Masson staining. The distribution of EdU-labeled ADSCs in healed wound tissue on PID 28 was observed by inverted fluorescence microscope. The expressions of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) of healed wound tissue on PID 7, 14, and 21 were detected by enzyme-linked immunosorbent assay. Data were processed with analysis of variance of factorial design and paired samples t test. RESULTS: (1) The wound healing time of rabbits in ADSCs group was (19.5±1.1) d post injury, which was significantly shorter than that in control group [(23.3±1.5) d, t=4.50, P<0.05]. On PID 7, wounds of rabbits in two groups were dry with no obvious exudation, and redness and swelling around wounds disappeared gradually, the wound healing rate of rabbits in ADSCs group was (15.1±2.4)%, which was close to that in control group [(13.7±3.1)%, t=1.20, P>0.05]. On PID 14, wounds of rabbits in ADSCs group were dry and scabbed obviously, and the wound healing rate was (73.1±5.7)%, while wounds of rabbits in control group were little scabbed with little exudation, and the wound healing rate was significantly lower than that in ADSCs group [(52.9±5.1)%, t=8.06, P<0.01]. On PID 21, wounds of rabbits in ADSCs group were generally healed, and the wound healing rate was (95.6±3.0)%, while a few wounds still existed in rabbits of control group, and the wound healing rate was significantly lower than that in ADSCs group [(78.6±3.7)%, t=9.73, P<0.01]. On PID 28, wounds of rabbits in two groups were totally healed with the healing rate of 100%, and texture and microvascular responses of healed wound tissue in ADSCs group were better than those in control group. (2) On PID 7, fibroblasts in healed wound tissue of rabbits in two groups were all increased, and there were little vascular and collagen fiber proliferation with no obvious differences. On PID 14, the number of fibroblasts in healed wound tissue of rabbits in ADSCs group was more than that in control group, and the collagen fibers in healed wound tissue of rabbits in ADSCs group were arranged in dense and uniform, while those in control group were sparse and irregular. On PID 21, skin layers were differentiated in healed wound tissue of rabbits in two groups, and collagen fibers in healed wound tissue of rabbits in ADSCs group were still denser than that in control group. On PID 28, newborn skin was well differentiated in healed wound tissue of rabbits in ADSCs group, which was better than that in control group. There were a lot of thick collagen fibers in healed wound tissue of rabbits in two groups, and EdU-labeled ADSCs were involved in skin texture of rabbits in ADSCs group. (3) The expressions of VEGF and EGF in healed wound tissue of rabbits in two groups were similar on PID 7 (with t values respectively 0.70 and 0.91, P values above 0.05), which in ADSCs group were significantly higher than those in control group on PID 14 and 21 (with t values from 2.85 to 4.81, P values below 0.01). CONCLUSIONS: The transplantation of ADSCs can promote the wound healing of skin deep partial-thickness scald wound of rabbit and shorten the wound healing time.

Reciprocal regulation of miRNAs and piRNAs in embryonic development.

MicroRNAs (miRNAs) and piwi-interacting RNAs (piRNAs) are two classes of small noncoding RNAs, both of which play roles in regulating tissue development. It is unknown whether these distinct classes of noncoding RNAs can regulate one another. Here we show that ectopic expression of miR-17 inhibited mouse fertility and early embryonic development. Specifically, we found that the piRNA amplification loop was repressed by miR-17-5p, leading to increased levels of transposition mutagenesis. This occurred by suppressing the amplification loop of piRNAs with an identical 5' sequence and by targeting Mili/Miwi2, an essential component of the piRNA amplification loop, and the DNA methyltransferase, Dnmt3a. We also found that increased levels of piRNAs could compete with miRNAs for target binding, resulting in increased expression of Dnmt3a and Mili. Increased Dnmt3a levels could in turn block miR-17-5p expression, while increased Mili expression could accelerate piRNA amplification and inhibit transposon generation, favoring embryonic development. We report for the first time the reciprocal regulation between miRNAs and piRNAs in mouse embryonic development.

Cyclin G2 inhibits epithelial-to-mesenchymal transition by disrupting Wnt/ß-catenin signaling.

Epithelial ovarian cancer (EOC) has the highest mortality rate among gynecological malignancies owing to poor screening methods, non-specific symptoms and limited knowledge of the cellular targets that contribute to the disease. Cyclin G2 is an unconventional cyclin that acts to oppose cell cycle progression. Dysregulation of the cyclin G2 gene (CCNG2) in a variety of human cancers has been reported; however, the role of cyclin G2 in tumorigenesis remains unclear. In this study, we investigated the function of cyclin G2 in EOC. In vitro and in vivo studies using several EOC-derived tumor cell lines revealed that cyclin G2 inhibited cell proliferation, migration, invasion and spheroid formation, as well as tumor formation and invasion. By interrogating cDNA microarray data sets, we found that CCGN2 mRNA is reduced in several large cohorts of human ovarian carcinoma when compared with normal ovarian surface epithelium or borderline tumors of the ovary. Mechanistically, cyclin G2 was found to suppress epithelial-to-mesenchymal transition (EMT), as demonstrated by the differential regulation of various EMT genes, such as Snail, Slug, vimentin and E-cadherin. Moreover, cyclin G2 potently suppressed the Wnt/ß-catenin signaling pathway by downregulating key Wnt components, namely LRP6, DVL2 and ß-catenin, which could be linked to inhibition of EMT. Taken together, our novel findings demonstrate that cyclin G2 has potent tumor-suppressive effects in EOCs by inhibiting EMT through attenuating Wnt/ß-catenin signaling.

Foxo3 activity promoted by non-coding effects of circular RNA and Foxo3 pseudogene in the inhibition of tumor growth and angiogenesis.

It has recently been shown that the upregulation of a pseudogene specific to a protein-coding gene could function as a sponge to bind multiple potential targeting microRNAs (miRNAs), resulting in increased gene expression. Similarly, it was recently demonstrated that circular RNAs can function as sponges for miRNAs, and could upregulate expression of mRNAs containing an identical sequence. Furthermore, some mRNAs are now known to not only translate protein, but also function to sponge miRNA binding, facilitating gene expression. Collectively, these appear to be effective mechanisms to ensure gene expression and protein activity. Here we show that expression of a member of the forkhead family of transcription factors, Foxo3, is regulated by the Foxo3 pseudogene (Foxo3P), and Foxo3 circular RNA, both of which bind to eight miRNAs. We found that the ectopic expression of the Foxo3P, Foxo3 circular RNA and Foxo3 mRNA could all suppress tumor growth and cancer cell proliferation and survival. Our results showed that at least three mechanisms are used to ensure protein translation of Foxo3, which reflects an essential role of Foxo3 and its corresponding non-coding RNAs.

miR-17 extends mouse lifespan by inhibiting senescence signaling mediated by MKP7.

Here we show that transgenic expression of miR-17 extends lifespan and inhibits cellular senescence. We propose that miR-17 acts as a critical regulator of cellular senescence and tumorigenesis. We demonstrate that miR-17 targets both ADCY5 and IRS1, upregulating the downstream signals MKP7, FoxO3, LC3B, and HIF1α, and downregulating mTOR, c-myc, cyclin D1, and JNK. Silencing either ADCY5 or IRS1 promoted autophagy and repressed cellular senescence and apoptosis. Repression of ADCY5 by miR-17 translocated membrane-bound RGS2 into the nucleus, promoting interactions of RGS2 with HIF1α and the MKP7 promoter, enhancing MKP7 transcription. ADCY5 repression by miR-17 also facilitated the translocation of EGFR and MKP7 from membrane into cytoplasmic and mitochondrial fractions. Importantly, we found that MKP7 inhibited senescence by dephosphorylating PRAS40 at Thr246 and mTOR at Ser2248, facilitating the interaction and loss of function of both molecules. Thus, the oncogenic miR-17 also acts pleiotropically to inhibit cellular senescence and extend longevity.

MicroRNA miR-93 promotes tumor growth and angiogenesis by targeting integrin-ß8.

It has been reported that the miR-106b∼25 cluster, a paralog of the miR-17∼92 cluster, possesses oncogenic activities. However, the precise role of each microRNA (miRNA) in the miR-106b∼25 cluster is not yet known. In this study, we examined the function of miR-93, one of the microRNAs within the miR-106b∼25 cluster, in angiogenesis and tumor formation. We found that miR-93 enhanced cell survival, promoted sphere formation and augmented tumor growth. Most strikingly, when miR-93-overexpressing U87 cells were co-cultured with endothelial cells, they supported endothelial cell spreading, growth, migration and tube formation. In vivo studies revealed that miR-93-expressing cells induced blood vessel formation, allowing blood vessels to extend to tumor tissues in high densities. Angiogenesis promoted by miR-93 in return facilitated cell survival, resulting in enhanced tumor growth. We further showed that integrin-ß8 is a target of miR-93. Higher levels of integrin-ß8 are associated with cell death in tumor mass and in human glioblastoma. Silencing of integrin-ß8 expression using small interfering RNA promoted cell proliferation, whereas ectopic expression of integrin-ß8 decreased cell growth. These findings showed that miR-93 promotes tumor growth and angiogenesis by suppressing, at least in part, integrin-ß8 expression. Our results suggest that inhibition of miR-93 function may be a feasible approach to suppress angiogenesis and tumor growth.

Malignant triton tumour of the parapharyngeal space: a case arising from the cervical sympathetic nerve.

BACKGROUND: Malignant triton tumour is a relatively rare, aggressive sarcoma comprising both malignant schwannoma cells and malignant rhabdomyoblasts. Malignant triton tumour of the parapharyngeal space is exceptionally rare, with only one case being described in the literature. Malignant triton tumour of the cervical sympathetic nerve has not previously been reported. METHODS: We report a case involving the parapharyngeal space and arising from the cervical sympathetic nerve, and we review the management and outcome of the previous case reported in this rare location. CONCLUSIONS: The parapharyngeal space is a unique location. Owing to this specific localisation, adjuvant therapy in addition to complete resection may be important in the treatment of malignant triton tumour in this rare location.

Fixed-dose pegfilgrastim is safe and allows neutrophil recovery in patients with non-Hodgkin's lymphoma.

Twenty-nine patients with non-Hodgkin's lymphoma received a single subcutaneous injection of 6 mg pegfilgrastim approximately 24 h after the start of CHOP chemotherapy. The safety of pegfilgrastim in this patient population was determined by reports of adverse events. The pharmacokinetics of pegfilgrastim were characterized and the duration of grade 4 neutropenia, time to absolute neutrophil count (ANC) recovery to > or = 2.0 x 10(9)/l, neutrophil nadir, and incidence of febrile neutropenia were determined in the first 21-day chemotherapy cycle. The incidence of grade 4 neutropenia in cycle 1 was 43% with a mean (SD) duration of grade 4 neutropenia value of 1.0 (1.4) day. No apparent relationship between the duration of grade 4 neutropenia and body weight was observed. The median [quartiles] time to ANC recovery was 10 [9, 11] days. The incidence of febrile neutropenia was 11%. No unexpected adverse events were reported and no patient developed antibodies to pegfilgrastim. Serum concentration of pegfilgrastim reached a maximum (median [quartiles]) of 128 [58, 159] ng/ml at approximately 24 h after administration, and was followed by a second smaller peak (median [quartiles]) of 10.6 [3.0, 20.5] ng/ml at the time of the neutrophil nadir. After the second peak, concentration of pegfilgrastim declined linearly with a median terminal half-life of approximately 42 h.

Pegfilgrastim, a sustained-duration form of filgrastim, significantly improves neutrophil recovery after autologous marrow transplantation in rhesus macaques.

Daily administration of filgrastim decreases the duration of severe neutropenia in the clinical setting. A sustained-duration form of filgrastim, pegfilgrastim, significantly reduces scheduling protocols to a single injection per chemotherapy cycle while maintaining therapeutic efficiency. We examined the ability of a single injection of pegfilgrastim to significantly improve neutrophil recovery following autologous bone marrow transplantation (AuBMT) in rhesus macaques. On day 1, postmyeloablation (920 cGy x-irradiation) and AuBMT, animals received either 0.1% autologous serum for 18 consecutive days (n=13), or single doses of pegfilgrastim via the subcutaneous (s.c.) or intravenous (i.v.) route (300 or 100 micro g/kg), or a single dose of filgrastim at 300 micro g/kg via the s.c. or i.v. route, or filgrastim at 10 micro g/kg via the s.c. route (n=4) on a daily basis (range=days 12-17). Pharmacokinetic parameters and neutrophil recovery were assessed. A single dose of pegfilgrastim via the i.v. or s.c. route was as effective as daily filgrastim administration, resulting in significant improvement of neutrophil recovery after myeloablation and ABuMT. Effective pegfilgrastim plasma concentrations were maintained in neutropenic animals until after the onset of hematopoietic recovery. Enhanced pharmacokinetics in AuBMT cohorts are consistent with self-regulating, neutrophil-mediated clearance.

Comparable efficacy and safety profiles of once-per-cycle pegfilgrastim and daily injection filgrastim in chemotherapy-induced neutropenia: a multicenter dose-finding study in women with breast cancer.

BACKGROUND: Neutropenia is common in patients receiving myelotoxic chemotherapy. Pegfilgrastim, a sustained-duration filgrastim is a once-per-cycle therapy for prophylactic neutrophil support. PATIENTS AND METHODS: Women, treated with four cycles of doxorubicin/docetaxel chemotherapy every 21 days, received pegfilgrastim or filgrastim 24 h after chemotherapy as a single subcutaneous injection per chemotherapy cycle (pegfilgrastim 30, 60 or 100 microg/kg) or daily subcutaneous injections (filgrastim 5 microg/kg/day). Safety, efficacy and pharmacokinetics were analyzed. RESULTS: The incidence of grade 4 neutropenia in cycle 1 was 95, 90 and 74%, in patients who received pegfilgrastim 30, 60 and 100 microg/kg, respectively, and 76% in patients who received filgrastim. Mean duration of grade 4 neutropenia in cycle 1 was 2.7,2 and 1.3 days for doses of pegfilgrastim, and 1.6 days for filgrastim. The pharmacokinetics of pegfilgrastim were non-linear and dependent on both dose and neutrophil count. Pegfilgrastim serum concentration was sustained until the neutrophil nadir occurred then declined rapidly as neutrophils started to recover, consistent with a self-regulating neutrophil-mediated clearance mechanism. The safety profiles of pegfilgrastim and filgrastim were similar. CONCLUSIONS: A single subcutaneous injection of pegfilgrastim 100 microg/kg provided neutrophil support and a safety profile comparable to daily subcutaneous injections of filgrastim during multiple chemotherapy cycles.

Identification of the motif in versican G3 domain that plays a dominant-negative effect on astrocytoma cell proliferation through inhibiting versican secretion and binding.

This study was designed to investigate the mechanisms by which mutant versican constructs play a dominant-negative effect on astrocytoma cell proliferation. Although a mini-versican or a versican G3 construct promoted growth of U87 astrocytoma cells, a mini-versican lacking epidermal growth factor (EGF) motifs (versicanDeltaEGF) and a G3 mutant (G3DeltaEGF) exerted a dominant-negative effect on cell proliferation. G3DeltaEGF-transfected cells formed smaller colonies, arrested cell cycle at G(1) phase, inhibited expression of cell cycle proteins cdk4 and cyclin D1, and contained multiple nucleoli. In cell surface binding assays, G3 products expressed in COS-7 cells and bacteria bound to U87 cell surface. G3DeltaEGF products exhibited decreased binding activity, but higher levels of G3DeltaEGF products were able to inhibit the binding of G3 to the cell surface. G3DeltaEGF expression inhibited secretion of endogenous versican in astrocytoma cells and also inhibited the secretion of mini-versican in COS-7 cells co-transfected with the mini-versican and G3DeltaEGF constructs. The effect seems to depend on the expression efficiency of G3DeltaEGF, and it occurred via the carbohydrate recognition domain.