RESUMO
BACKGROUND: Estrogen receptor alpha (ERa/ESR1) expression is regulated by alternative splicing. Its most frequently detectable exon7 skipping isoform (ERaD7) is a dominant negative variant. Elevated expression of ERaD7 was already detected in endometrial cancer (EC), while its potential prognostic significance has not been characterized so far. Exon7 contains potential binding sites for the two functional splicing regulatory opponents, HNRNPG and HTRA2-BETA1 known to trigger opposite effects on EC outcome. This study served to elucidate the influence of HNRNPG and HTRA2-BETA1 on ERa exon7 splicing regulation and the impact of ERaD7 concentration on type 1 EC outcome. METHODS: Functional in vitro experiments for HNRNPG and HTRA2-BETA1 in regard to the regulatory impact on endogenous and exogenous ERaD7 splicing were performed. Additionally, real-time PCR determined mRNA levels of ERaD7, HNRNPG and HTRA2-BETA1 in 116 type 1 EC patients. RESULTS: HNRNPG and HTRA2-BETA1 were found to be specific regulators of ERa exon7 splicing. While HTRA2-BETA1 promoted exon7 inclusion, HNRNPG antagonized this effect by inducing exon7 skipping (p = 0.004). ERaD7 was detected in 71 out of 116 type 1 EC specimens. Statistical analyses revealed an inverse correlation between ERaD7 mRNA levels and tumor grading (p = 0.029), FIGO stage (p = 0.033) as well as lymph node metastases (p = 0.032), respectively. Furthermore, higher ERaD7 expression could be correlated to an improved disease-specific survival (p = 0.034). CONCLUSIONS: Our study demonstrates antagonistic regulatory effects of HNRNPG and HTRA2-BETA1 on ERa exon7 splicing with potential impact on type 1 EC clinical outcome due to the consecutively variable expression levels of the ERa isoform D7.
Assuntos
Neoplasias do Endométrio/genética , Neoplasias do Endométrio/patologia , Receptor alfa de Estrogênio/genética , Ribonucleoproteínas Nucleares Heterogêneas/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Proteínas de Ligação a RNA/metabolismo , Processamento Alternativo , Sítios de Ligação , Linhagem Celular Tumoral , Neoplasias do Endométrio/metabolismo , Receptor alfa de Estrogênio/química , Receptor alfa de Estrogênio/metabolismo , Éxons , Feminino , Células HeLa , Ribonucleoproteínas Nucleares Heterogêneas/genética , Humanos , Proteínas do Tecido Nervoso/genética , Prognóstico , Proteínas de Ligação a RNA/genética , Fatores de Processamento de Serina-ArgininaRESUMO
OBJECTIVE: To retrospectively evaluate the efficacy of a new complementary mid-urethral sling surgery (Tong's hammock anterior, THA) in treatment of recurrent or persist stress urinary incontinence (SUI) in females after primary synthetic mid-urethral slings (MUSs). METHODS: THA was performed in 27 females with recurrent or persist SUI after primary MUSs from June 2005 and July 2010. These patients were followed up for one year, and clinical data including main complaints, operation duration, blood loss, efficacy and complications were reviewed. RESULTS: All 27 SUI patients were treated with THA surgery, a trans-vaginal mid-urethral sling on the descending pubic ramus. The average operation time was 39 min (range: 25-70 min), average blood loss was 70 ml (range: 20-120 ml). After urinary catheter removal, all patients could micturate and their average residual urine was 25.2 ml (range: 0-80 ml). The average hospital stay was 4.7 days (rage: 3-7 days). SUI symptom was persistent in 2 patients after THA surgery and the effective rate reached 92.5%. At 3 months, 6 months and 1 year after surgery, the effective rate was 92.5% (25/27), 92% (23/25) and 87.5% (21/24), respectively. 6 months after THA surgery, 2 were lost to follow up; 1 had recurrent SUI at 1 year and 1 had mesh erosion, 1 died of other diseases, and operative complications were absent after surgery. CONCLUSIONS: THA surgery is an effective method for treating recurrent or persistent SUI after primary MUSs. It is cheap, efficient, and easy to handle.
RESUMO
HnRNP G is a member of heterogeneous nuclear ribonucleoprotein (hnRNP) family with potent tumor suppressive activities. Human transformer-2-beta1 (hTra2-beta1) belongs to the arginine-serine rich like proteins and is found over-expressed in various human cancers. It was recently shown that hnRNP G and hTra2-beta1 exert antagonistic effects on alternative splicing. In our study we explored the impact of these two factors in tumor biology of endometrial cancer (EC). EC tissues (n = 139) were tested for hnRNP G and hTra2-beta1 expression on mRNA level by real time PCR and on protein level by immunohistochemistry. HTra2-beta1 mRNA level was found being induced in advanced International Federation of Gynecology and Obstetrics (FIGO) stages (p = 0.016). HnRNP G protein nuclear expression was found more prominent in patients without distant organ metastases (p = 0.033) and in FIGO Stages I/II group (p < 0.001). HTra2-beta1 protein nuclear levels were elevated in poorly differentiated (p = 0.044) and lymph node metastases (p = 0.003) cancers. Kaplan-Meier survival curves revealed that elevated hnRNP G mRNA (p = 0.029) and protein (p = 0.022) levels were associated with a favorable patient outcome. Multivariate Cox-regression analyses identified nuclear hnRNP G level [hazard ratio (HR) 0.468, p = 0.026) as well as hTra2-beta1 level (hazard ratio 5.760, p = 0.004) as independent prognostic factors for EC progression-free survival. Our results indicate that the antagonistic functional effects of hnRNP G and hTra2-beta1 on alternative splicing correlate directly to their opposite clinical effects on EC patient outcome.