Cytoplasmic localization of IRF5 induces Wnt5a/E-cadherin degradation and promotes gastric cancer cells metastasis.

IRF5, a nucleoplasm shuttling protein, is a pivotal transcription factor regulating immune system activity. It's well known that immunosuppression is involved in the development of gastric cancer. However, no data exist for the expression and function of IRF5 in gastric cancer. This study demonstrated that IRF5 was cytoplasm-enriched in gastric cancer cells. IRF5 promoted gastric cancer cell migration, which involved the inhibition of Wnt5a and E-cadherin proteins expression. IRF5 (LA) localized in nucleus had no significant effect on Wnt5a and E-cadherin expressions, while mutation of IRF5 (ΔNLS), which prevents IRF5 nuclear translocation, had more impact on these inhibitory effects. In addition, degradation rates of both Wnt5a and E-cadherin were enhanced by resiquimod, an IRF5 agonist. Further in vivo experiments indicated that IRF5 knockout of gastric cancer cells repressed their pulmonary metastasis in nude mice. Finally, the expression and clinical significance of IRF5 were analyzed using gastric cancer tissue microarrays, which suggested that the expression of IRF5 varied procedurally in different progressive stages of gastric cancer. Our data revealed that IRF5 cytoplasmic localization were associated with Wnt5a and E-cadherin degradation and gastric cancer cell metastasis. Inhibiting IRF5 expression and/or its cytoplasmic localization may provide a novel target for gastric cancer therapy.

[The correlation between sperm DNA damage and IVF-ET clinical pregnancy outcomes in different BMI population with normal routine semen examination].

OBJECTIVE: To analyze the correlation between sperm DFI, HDS and IVF-ET pregnancy outcomes in different BMI populations with normal routine semen examination. METHODS: The clinical data of 199 cycles of IVF-ET were retrospectively analyzed. Sperm chromatin structure analysis based on flow cytometry was used to detect sperm DFI and HDS. The correlation between sperm DFI, HDS and pregnancy outcome of IVF-ET were analyzed. RESULTS: The sperm DFI was negatively correlated with IVF-ET pregnancy in overweight (24.0 kg/m2≤BMI<28.0 kg/m2) population (OR=0.935, P=0.043). In the normal BMI group (18.5 kg/m2≤BMI < 24.0 kg/m2), the clinical pregnancy outcome of IVF-ET was not significantly correlated with sperm DFI, and was negatively correlated with male age (OR=0.744, P=0.020). In the obese population (BMI ≥ 28.0 kg/m2) , there was no significant correlation between the clinical pregnancy outcome of IVF-ET and sperm DNA fragmentation index (DFI) , but a negative correlation with male BMI (OR = 0.779, P = 0.043). CONCLUSION: The male BMI affected the correlation between sperm DFI and IVF-ET pregnancy outcomes: ①Sperm DFI was only associated with IVF-ET clinical pregnancy outcome in the overweight population; ② In normal BMI and obese populations, male age and male BMI were important factors affecting IVF-ET clinical pregnancy outcome respectively; ③No correlation was found between sperm HDS and IVF-ET pregnancy outcomes.

Six-Year Follow-Up of a Rare Bifid Talus and Eight-Toed Central Polydactyly: A Case Report.

Eight-toed central polydactyly is a rare congenital foot deformity and no other case with a bifid talus has been reported in the literature. We present a 6-year follow-up of a male child who had eight-toed central polydactyly with a duplicate cuneiform bone and bifid talus in his right foot. During preoperative planning, CT scans were conducted to evaluate the duplicate tarsals and to assist in reaching surgical decisions. In 2013, when the child was 1 year and 8 months old, the fourth, fifth and sixth phalanges and metatarsals as well as the duplicate cuneiform bone were excised. A portion of the malformed talus was also resected. This case report discusses functional and aesthetic outcomes after 6 years and provides an analysis on relevant reconstructive follow-up practices.

[Assisted reproductive technology for male patients with end-stage renal disease: A case-based discussion on strategies and ethical issues].

OBJECTIVE: To explore normalized and reasonable strategies of assisted reproductive technology (ART) for patients with end-stage renal disease (ESRD) under ethical supervision based on the experience with a case of ART for an ESRD male. METHODS: A male patient with ESRD successfully fathered a child through in vitro intracytoplasmic sperm injection (ICSI) in our center. We performed an epidemiological analysis, reviewed the relevant literature and explored the feasibility, ethical issues and strategies of ART for male patients with ESRD. RESULTS: ESRD affected the reproductive hormone levels, sperm quality and erectile function of the patient. Considering the contradictions between the reproductive right and the uncertainty of disease prognosis of the patient and the health of the offspring and his wife, we comprehensively evaluated the physical and mental conditions of the patient, obtained the informed consent, submitted the case to the Ethics Committee of Reproductive Medicine. CONCLUSIONS: With respect to ART for ESRD patients, importance should be attached to their rights of reproduction and choice of reproductive technology. In the process of ART, the physical conditions of the patient ought to be evaluated comprehensively and rigorously, and the related ethical principles followed strictly.

Associations between various possible promoter polymorphisms of MMPs genes and endometriosis risk: a meta-analysis.

BACKGROUND: Endometriosis is a common, benign gynecological disorder affecting life quality of reproductive-aged women. Several polymorphisms in the promoter regions of the MMPs genes have been reported that were related to endometriosis risk. However, there are many contradictory conclusions, and no meta-analysis focused on this association systematically. OBJECTIVES: To evaluate the associations between various possible polymorphisms of MMPs genes and endometriosis risk, and confirm which kinds of MMPs genetic polymorphisms are associated with endometriosis risk, in order to identify the etiology and pathogenesis of endometriosis and the potential effective markers for predicting the predisposition to endometriosis. SEARCH STRATEGY: An exhaustive electronic literature search was conducted, using keywords MMP, endometriosis and SNP, in English and Chinese. SELECTION CRITERIA: All eligible case-control studies by written in English or Chinese of the associations of MMPs polymorphisms with endometriosis risk, which had sufficient data for examining an odds ratio (OR) with 95% confidence interval (CI), were identified up to March 1, 2015. DATA COLLECTION AND ANALYSIS: A total of 1833 patients and 2190 controls from 12 studies were included. Allele frequency differences between cases and controls were performed with the use of odds ratios (ORs) and their respective 95% confidence intervals (CIs) for five genetic models. MAIN RESULTS: For MMP-1 -1607 1G>2G (rs1799750) polymorphism, significant associations were observed both in overall comparison and subgroup analyses based on the stage of endometriosis, ethnicity of each study population and method of genotyping under four genetic models. In contrast, for MMP-2 15918 T>C (rs243847), MMP-2 -753 C>T (rs2285053), MMP-7 -181 A>G (rs11568818), MMP-9 -1562 C>T (rs3918242) and MMP-9 R279Q (rs17576) polymorphisms, no association was found in overall comparison, but in subgroup analyses based on source of control, stage of endometriosis, or ethnicity. CONCLUSIONS: MMP-1 -1607 1G>2G polymorphism might modulate risk of endometriosis, so does the MMP-2 15918 T>C, MMP-2 -753 C>T, MMP-7 -181 A>G, MMP-9 -1562 C>T and MMP-9 R279Q polymorphisms in some subgroups.

[Application of calcium ionophore A23187 in ICSI for globozoospermia: A report of 2 cases and review of the literature].

OBJECTIVE: To investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men. METHODS: We collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease. RESULTS: Quality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group. CONCLUSION: Calcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

EGF-reduced Wnt5a transcription induces epithelial-mesenchymal transition via Arf6-ERK signaling in gastric cancer cells.

Wnt5a, a ligand for activating the non-canonical Wnt signaling pathway, is commonly associated with Epithelial-to-mesenchymal transition (EMT) in cancer cell metastasis. Here, we show that downregulation of Wnt5a mRNA and protein by EGF is necessary for EGF-induced EMT in gastric cancer SGC-7901 cells. To further explore the mechanisms, we investigated the effect of EGF signaling on Wnt5a expression. EGF increased Arf6 and ERK activity, while blockade of Arf6 activation repressed ERK activity, up-regulated Wnt5a expression and repressed EMT in response to EGF. We also demonstrate that EGF inactivated Wnt5a transcription by direct recruitment of ERK to the Wnt5a promoter. On the other hand, inhibition of ERK phosphorylation resulted in decreased movement of ERK from the cytoplasm to the nucleus, following rescued Wnt5a mRNA and protein expression and favored an epithelial phenotype of SGC-7901 cells. In addition, we notice that kinase-dead, nuclear-localised ERK has inhibitory effect on Wnt5a transcription. Analysis of gastric cancer specimens revealed an inverse correlation between P-ERK and Wnt5a protein levels and an association between Wnt5a expression and better prognosis. These findings indicate that Wnt5a is a potential suppressor of EMT and identify a novel Arf6/ERK signaling pathway for EGF-regulated Wnt5a expression at transcriptional level of gastric cancer cells.

GEP100 regulates epidermal growth factor-induced MDA-MB-231 breast cancer cell invasion through the activation of Arf6/ERK/uPAR signaling pathway.

GEP100, a guanine nucleotide exchanging factor (GEF) for Arf6, plays a pivotal role in promoting breast cancer cell invasion both in vitro and in vivo. However, the precise mechanism for GEP100-mediated cell invasion is still poorly understood. In this study, we found that down-regulation of endogenous GEP100 in MDA-MB-231 cells significantly inhibited EGF-induced cell invasion, which was rescued by over-expression of ectopic GEP100. EGF increased Arf6 activity, ERK phosphorylation, and uPAR expression in a time dependent manner. Additionally, blocking Arf6 with Arf6 siRNA largely abolished EGF-induced cell invasion. GEP100 siRNA or Arf6 siRNA suppressed EGF-induced ERK activity and uPAR expression. Furthermore, blocking ERK signaling with U0126, a specific inhibitor for MEK, markedly inhibited EGF-induced uPAR expression and consequently cell invasion. Inhibition of uPAR expression by uPAR siRNA also significantly abolished EGF-induced cell invasion. Taken together, this study illustrates that GEP100 regulates an Arf6/ERK/uPAR signaling cascade in EGF-induced breast cancer cell invasion. These findings could provide a rationale for designing new therapies based on inhibition of breast cancer metastasis.

Rab35 is required for Wnt5a/Dvl2-induced Rac1 activation and cell migration in MCF-7 breast cancer cells.

The small GTPases regulate many major biological processes in both tumorigenesis and tumor progression such as cell survival, actin cytoskeleton organization, cell polarity and movement. Wnt5a, a non-canonical Wnt family member, is implicated in the activation of small GTPases in breast cancer. We previously demonstrated that Wnt5a signaling stimulates the migration of breast cancer cells MDA-MB-231 via activating RhoA. However, we found here that RhoA activation was not enhanced by Wnt5a in breast cancer cells MCF-7. The conflicting results prompted us to further probe novel small GTPases in response to Wnt5a and investigate the mechanisms whereby cell migration is regulated. We showed here that Wnt5a dose dependently activated Dvl2, Rab35 and Rac1 and subsequently promoted the migration of MCF-7 cells, which was, however, abolished by knocking down Wnt5a expression via small interfering RNA (siRNA) transfection. Dvl2 siRNA significantly decreased background and Wnt5a-induced Rab35/Rac1 activation and, consequently, cell migration. Rab35 short hairpin RNA (shRNA) remarkably inhibited background and Wnt5a-induced Rac1 activation and cell migration. Additionally, blockade of Rac1 activation with Rac1 siRNA suppressed background and Wnt5a-induced cell migration. Co-immunoprecipitation and immunofluorescence assays showed that Dvl2 bound to Rab35 in mammalian cells. Taken together, we demonstrated that Wnt5a promotes breast cancer cell migration via the Dvl2/Rab35/Rac1 signaling pathway. These findings implicate Wnt5a signaling in regulating small GTPases, which could be targeted for manipulating breast cancer cell migration.

PI3K/Akt-dependent phosphorylation of GSK3ß and activation of RhoA regulate Wnt5a-induced gastric cancer cell migration.

Wnt5a, a non-transforming Wnt family member, plays complicated roles in oncogenesis and cancer metastasis. However, Wnt5a signaling in gastric cancer progression remains poorly defined. In this study, we found that Wnt5a dose-dependently stimulated the migration of human gastric cancer cells (SGC-7901), with the maximal effect at 100 ng/mL, via enhancing phosphorylation of PI3K/Akt and GSK3ß and activating RhoA. Pharmaceutical inhibition of PI3K with LY294002 or Akt siRNA significantly decreased Wnt5a-induced GSK3ß phosphorylation and consequently cell migration. Additionally, GSK3ß siRNA remarkably inhibited Wnt5a-induced RhoA activation, stress fiber formation and cell migration. Analogously, pre-treatment with LiCl, which induced phosphorylation of GSK3ß at Ser9, increased Wnt5a-induced cell migration. Finally, ectopic expression of dominant negative RhoA (N19) suppressed Wnt5a-induced cell migration. Taken together, we demonstrated for the first time that Wnt5a promoted gastric cancer cell migration via the PI3K/Akt/GSK3ß/RhoA signaling pathway. These findings could provide a rationale for designing new therapy targeting gastric cancer metastasis.

Dvl2-dependent activation of Daam1 and RhoA regulates Wnt5a-induced breast cancer cell migration.

BACKGROUND: The Dishevelled (Dvl) and Dishevelled-associated activator of morphogenesis 1 (Daam1) pathway triggered by Wnt5a regulates cellular polarity during development and tissue homoeostasis. However, Wnt5a signaling in breast cancer progression remains poorly defined. METHODOLOGY/PRINCIPAL FINDINGS: We showed here that Wnt5a activated Dvl2, Daam1 and RhoA, and promoted migration of breast cancer cells, which was, however, abolished by Secreted Frizzled-related protein 2 (sFRP2) pretreatment. Dominant negative Dvl2 mutants or Dvl2 siRNA significantly decreased Wnt5a-induced Daam1/RhoA activation and cell migration. Ectopic expression of N-Daam1, a dominant negative mutant, or Daam1 siRNA remarkably inhibited Wnt5a-induced RhoA activation, stress fiber formation and cell migration. Ectopic expression of dominant negative RhoA (N19) or C3 exoenzyme transferase, a Rho inhibitor, decreased Wnt5a-induced stress fiber formation and cell migration. CONCLUSIONS/SIGNIFICANCE: Taken together, we demonstrated for the first time that Wnt5a promotes breast cancer cell migration via Dvl2/Daam1/RhoA.

Activation of Rac1-PI3K/Akt is required for epidermal growth factor-induced PAK1 activation and cell migration in MDA-MB-231 breast cancer cells.

Epidermal growth factor (EGF) may increase cell motility, an event implicated in cancer cell invasion and metastasis. However, the underlying mechanisms for EGF-induced cell motility remain elusive. In this study, we found that EGF treatment could activate Ras-related C3 botulinum toxin substrate 1 (Rac1), PI3K/Akt and p21-actived kinase (PAK1) along with cell migration. Ectopic expression of PAK1 K299R, a dominant negative PAK1 mutant, could largely abolish EGF-induced cell migration. Blocking PI3K/Akt signalling with LY294002 or Akt siRNA remarkably inhibited both EGF-induced PAK1 activation and cell migration. Furthermore, expression of dominant-negative Rac1 (T17N) could largely block EGF-induced PI3K/Akt-PAK1 activation and cell migration. Interestingly, EGF could induce a significant production of ROS, and N-acetyl-L-cysteine, a scavenger of ROS which abolished the EGF-induced ROS generation, cell migration, as well as activation of PI3K/Akt and PAK, but not Rac1. Our study demonstrated that EGF-induced cell migration involves a cascade of signalling events, including activation of Rac1, generation of ROS and subsequent activation of PI3K/Akt and PAK1.