The Molecular Mechanism Of GC1qR In Apoptosis Of Ovarian Cancer Cells Induced By Paclitaxel/Carboplatin

Part I The study of mechanism of gC1qR gene on ovarian cancer cell apoptosis.【Objective】To investigate the mechanism of the receptor of globular C1q(gC1qR)gene on the apoptosis of human ovarian cancer cells.【Methods】Construction of recombinant plasmid pc DNA3.1-gC1qR,then transfected into human ovarian cancer cells SKOV3,the level of gC1qR gene was measured by Real time PCR and Western blot,the apoptosis of ovarian cancer cell was assessed by flow cytometry.The intracellular ROS was estimated via the fluorescence of 2,,7,-dichlorodihydrofluorescein diacetate(H2DCFDA),the mitochondrial membrane potential was tested using a tetrechloro-tetraethylb-enzimidazol carbocy-anine iodide(JC-1)probe,and the intracellular Ca2+ was assessed via Fluo-3/AM.【Results】The ovarian cancer cells transfected with pc DNA3.1-gC1qR could induce the cell apoptosis,and showed significant cell apoptosis morphology.The intracellular ROS and Ca2+ were obviously increased,and the mitochondrial membrane potential was obviously decreased.【Conclusions】These results indicate that gC1qR gene induced mitochondria dysfunction,which induced the apoptosis of ovarian cancer cells.Part II The molecular mechanism of Paclitaxel/Carboplatin induced the apoptosis of ovarian cancer cells【Objective】To investigate the molecular mechanism of Paclitaxel/ Carboplatin induced the apoptosis of ovarian cancer cells.【Methods】Construction of recombinant plasmid gC1qR si RNA,then transfected into human ovarian cancer cells SKOV3,negative si RNA group and α-lipoic acid group were considered negative control and positive control corresponding.The level of gC1qR gene was measured by Real time PCR and Western blot,the apoptosis of ovarian cancer cell was assessed by flow cytometry.The intracellular ROS was estimated via the fluorescence of H2 DCFDA,the mitochondrial membrane potential was tested using a JC-1 probe,and the intracellular Ca2+ was assessed via Fluo-3/AM.【Results】The SKOV3 treated with Paclitaxel/ Carboplatin + gC1qR si RNA and Paclitaxel/ Carboplatin + α-lipoic acid,the number of apoptosis cells were decreased,the level of ROS and intracellular Ca2+ were apparently decreased,at same time,the mitochondrial membrane potential was obviously increased.【Conclusions】These results indicate that gC1qR-induced mitochondria dysfunction was involved in Paclitaxel/ Carboplatin-mediated the apoptosis of ovarian cancer cells.