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Purpose To investigate the expression of globular C1q receptor (gC1qR) in ovarian cancer and to explore its potential molecular mechanism.Methods Retrospective analysis was made on 48 ovarian cancer tissues and ovarian cancer cell line SKOV3.Real time PCR and Western blot analysis were applied to detect the levels of gC1qR mRNA and gC1qR protein expression.The abilities of SKOV3 cells proliferation activity and quantity,migration and apoptosis were respectively assessed by M'TT,transwell assay and flow cytometry.Besides,the intracellular ROS was estimated via the fluorescence of H2DCFDA,the mitochondrial membrane potential was tested using a JC-1 probe,and the intracellular Ca2+ was assessed via Fluo-3/AM.Results The expressions of gC1qR gene was obviously decreased in the group of ovarian cancer tissues when compared with normal ovarian tissues group (2.61 ±0.34 vs 7.32 ± 1.25,0.20 ± 0.02 vs 0.67-± 0.06,P ＜ 0.001).Overexpression of gC1qR gene could result in significant up-regulation of ovarian cancer cell apoptosis and down-regulation in proliferation and migration,and showed significant cell apoptosis morphology.Simultaneously,the intracellular ROS and Ca2+ were obviously increased,and the mitochondrial membrane potential was obviously decreased.Conclusion gC1qR gene may play an important role in the apoptosis of ovarian cancer cells.In this process,gC1qR gene can induce apoptosis of ovarian cancer cells by inducing mitochondrial dysfunction.
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[Abstract ] Objective Globular C1q receptor (gC1qR), a highly acidic receptor protein, is expressed in almost all mamma-lian cells in addition to exoerythrocytic, which can mediate a variety of biological responses.The study aimed to explore gC1qR expres-sion in cervical cancer and itd effects on the biological behaviors of human cervical cancer cells. Methods Retrospective analysis was made on 100 cervical tissue samples of patients in Nanjing Maternal and Child Health Hospital from August 2014 to April 2015, in-cluding 50 cervicitis tissues and 50 cervical cancer tissues.Immunohistochemical SP method was applied in the research of cervical cancer cell line C33a to detect the expression and the location of gC1qR in cervical tissues.Real-time PCR and Western blot analysis were respectively applied to detect the levels of gC1qR mRNA and gC1qR protein expression.Besides, the abilities of C33a cells mi-gration, invasion and apoptosis were respectively assessed by in vitro cell wound healing experiment, transwell assay and flow cytome-try. Results The expression of gC1qR gene was dramatically decreased in the group of cervical cancer tissues when compared with chronic cervictis group (2.18 ±0.37 vs 7.23 ±0.69, 0.27 ±0.09 vs 0.74 ±0.02, P cal cells by transmission electron microscope. Conclusion gC1qR expression might play an important role in inhibiting the invasion and migration of cervical cancer cells and inducing the apoptosis of cervical carcinoma cells, which provides new clues and potential targets for the treatment of cervical cancer in further research.
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El gC1qR, un receptor para el dominio globular del C1q, es una proteína multicompartamental y multifuncional, cuya intervención en el proceso de fecundación en humanos se ha demostrado recientemente. Los objetivos de esta investigación fueron describir la distribución de gC1qR y valorar su expresión en la membrana plasmática de espermatozoides bovinos antes y después del tratamiento con heparina. La capacitación de espermatozoides provenientes de semen descongelado bovino se indujo con una incubación con heparina y su efectividad se evaluó con la tinción CTC. Posteriormente, se realizaron ensayos de inmunofluorescencia indirecta con el anticuerpo monoclonal 60.11 (anti-gC1qR). El análisis de los datos demostró que el gC1qR es una molécula ubicada en la membrana plasmática de los espermatozoides bovinos que presentauna redistribución tras la capacitación, concentrándose en la región acrosomal: 175 espermatozoides en las alícuotas incubadas con heparina vs. 109 en el control (P<0,05; n=300) presentaron fluorescencia en la región acrosomal; esta distribución es semejante a la documentada en humanos. Igualmente, el nivel de expresión o de accesibilidad de gC1qR en el espermatozoide bovino aumentó tras la capacitación, hecho que se ha observado también en humanos. Estos resultados sugieren que el gC1qR podría participar en la interacción primaria entre el espermatozoide y el ovocito en bovinos.
The complement receptor gC1qR/p33, which recognizes the globular heads of C1q, is a multicompartmental and multifunctional protein and has been shown to play a role in reproduction in humans. The objective of this research was to determine the gC1qR distribution and expression on plasma membrane in bovine sperm before and after heparin treatment. Thus, capacitation of sperm derived from frozen-thawed bovine semen was induced through heparin incubation and its effectiveness was assessed with a CTC stain. Subsequently, indirect immunofluorescence assays were conducted with mAb (60.11, anti-gC1qR) to assess gC1qR distribution and expression. Data analysis demonstrated that gC1qR is expressed on the plasma membrane of bovine sperm. gC1qR showed a capacitation-related redistribution, migrating to the acrosome region: 175 sperm in heparin-incubated aliquots vs. 109 in control (P<0.05, n=300) showed fluorescence over the acrosome region. This distribution is similar to that reported in humans. Similarly, either gC1qR expression or its accessibility to antibodies increased after capacitation. This also correlates with what has been observed in humans. These results suggest that gC1qR could participate in primary sperm-oocyte interaction in bovines.