RESUMO
INTRODUCTION: Preeclampsia (PE) is a pregnancy-specific complication and it is related to insufficient extravillous trophoblast invasion. To date, the pathophysiology of PE has not yet been fully elucidated. Response gene to complement 32 (RGC32) is a novel cellular protein, and it plays important roles in the regulation of cell differentiation, angiogenesis, migration, and invasion. This study aimed to determine the RGC32 expression and function in human placentas and to explore the underlying mechanisms. METHODS: RGC32 expression in term placentas collected after cesarean section from pregnant women with PE and normal pregnant women was determined by real-time reverse transcriptase polymerase chain reaction (RT-PCR), Western blot, and immunohistochemistry. The effects of RGC32 expression on trophoblast invasion, migration, and the underlying mechanisms were studied in HTR8/SVneo cells. RESULTS: The messenger RNA (mRNA) and protein levels of RGC32 were significantly downregulated in preeclamptic placentas compared with normal controls (P < 0.05). RGC32 silencing significantly inhibited HTR8/SVneo cell migration and invasion (P < 0.001, respectively). These effects were associated with decreased activities and expression of matrix metalloproteinase (MMP)-2/9, and with the reduced phosphorylation level of Akt. DISCUSSION: RGC32 may play important roles in the pathophysiology of PE by directly affecting the invasion/migration of trophoblast.
Assuntos
Proteínas de Ciclo Celular/metabolismo , Regulação para Baixo , Proteínas Musculares/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Pré-Eclâmpsia/metabolismo , Trofoblastos/metabolismo , Adulto , Western Blotting , Proteínas de Ciclo Celular/antagonistas & inibidores , Proteínas de Ciclo Celular/genética , Linhagem Celular , Movimento Celular , Cesárea , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Imuno-Histoquímica , Proteínas Musculares/antagonistas & inibidores , Proteínas Musculares/genética , Proteínas do Tecido Nervoso/antagonistas & inibidores , Proteínas do Tecido Nervoso/genética , Placentação , Pré-Eclâmpsia/patologia , Gravidez , Interferência de RNA , RNA Mensageiro/metabolismo , RNA Interferente Pequeno , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Trofoblastos/patologiaRESUMO
A sophisticated immunological regulation between decidual stromal cells (DSC) and monocytes and macrophages is essential for the successful symbiosis of the mother and her fetus, but the mechanisms remain incompletely understood. The mRNA and proteins of B lymphocyte stimulator (BAFF, also known as BLys) and its receptor, BAFF-R (also known as BR3, CD268 or TNFRSF17), have been detected in both first-trimester and term placentas, but whether BAFF or BAFF-R participates in the cross-talk between DSC and monocytes and macrophages in the first-trimester pregnancy has not been described. This study found that purified DSC extensively shed BAFF-R and that polyinosinic:polycytidylic acid (poly(I:C); a synthetic toll-like receptor (TLR) 3 agonist) dramatically up-regulated BAFF-R secretion, suggesting that release of these soluble proteins was an inherent property of DSC and its induction might have relevance to TLR-3-mediated signal transduction. When monocytes were cultured with the supernatants of resting DSC or poly(I:C)-treated DSC, the proliferation of CD14(+)HLA-DR(+) monocytes (P=0.025 and 0.045) and the secretion levels of tumour necrosis factor α (P=0.035 and 0.031) and interleukin 6 (P=0.021 and 0.035) were significantly increased after the BAFF-R was blocked. Soluble BAFF-R may play inhibitory roles in monocytes and macrophages.
Assuntos
Receptor do Fator Ativador de Células B/metabolismo , Receptor do Fator Ativador de Células B/farmacologia , Decídua/metabolismo , Macrófagos/efeitos dos fármacos , Monócitos/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Decídua/citologia , Decídua/efeitos dos fármacos , Feminino , Humanos , Técnicas In Vitro , Interleucina-6/metabolismo , Macrófagos/citologia , Macrófagos/metabolismo , Monócitos/citologia , Monócitos/metabolismo , Poli I-C/metabolismo , Poli I-C/farmacologia , Gravidez , Primeiro Trimestre da Gravidez , Células Estromais/citologia , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Receptor 3 Toll-Like/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Human peripheral blood monocytes are a heterogeneous population, including CD14(+) CD16(-) 'classical' monocytes and CD14(+) CD16(+) 'proinflammatory' monocytes. CD16(+) monocytes are expanded in various inflammatory conditions. However, little is known about the CD14(+) CD16(+) monocytes in patients with breast cancer. We detected CD14(+) CD16(+) monocytes in 96 patients with breast cancer and 54 control subjects using flow cytometry. Receiver-operating characteristic (ROC) curve analysis was used to determine the feasibility of CD14(+) CD16(+) monocytes as an indicator for diagnosis of breast cancer. We found that the frequency of CD14(+) CD16(+) monocytes showed a significantly greater increase in breast cancer patients than in controls (16·96% versus 10·84%, P < 0·0001). The area under the ROC curve for CD14(+) CD16(+) monocytes was 0·805 [95% confidence interval (95% CI): 0·714-0·877, P = 0·0001]. Furthermore, the levels of CD16(+) monocytes were significantly negatively associated with the tumour size and pathological staging. In vitro, we showed that CD14(+) CD16(+) monocytes were expanded significantly when the purified CD14(+) monocytes were exposed to Michigan Cancer Foundation (MCF)-7 cells-conditioned medium (MCF-CM) or, separately, to monocyte chemotactic protein 1 (MCP-1). Neutralizing antibodies against MCP-1 inhibited the expansion of CD14(+) CD16(+) monocytes by MCF-CM. Collectively, our findings indicated that MCP-1 can expand CD14(+) CD16(+) monocytes in patients with breast cancer. Furthermore, the CD14(+) CD16(+) monocyte may be a useful indicator in early diagnosis of breast cancer.
