Mechanism Of Apoptosis Of Cervical Cancer Cells Induced By GC1qR Gene

PartⅠExpression of gC1qR brings the changes of the biological behaviors of human cervical cancer cellsAims:Globular C1q receptor(gC1qR),a highly acidic receptor protein,can mediate a variety of biological responses.Our study aims to study expression of gC1qR in cervical cancer and the effects it brings on the biological behaviors of human cervical cancer cells.Methods:Focusing on cervical cancer cell line C33a,SiHa,CRL2614,we used Real-time PCR and Western blot analysis to detect the levels of gC1qR mRNA and protein expression in cervical tissues and cells.Besides,WST,transwell assay and flow cytometry were used to assess the abilities of cells migration,invasion and apoptosis.Results:gC1qR gene expression was dramatically decreased in the group of cervical cancer tissues and cells.Meanwhile,in cervical cancer cells that were transfected with gC1qR vector displayed significant up-regulation of celluer apoptosis and down-regulation in migration and invasion.Conclusions:The data suggested that gC1qR might play an important role in inhibiting the invasion and migration of cervical cancer cells,and inducing the apoptosis of cervical carcinoma cells.So it is expected to be an effectively possible target for the treatment of cervical cancer in the further research.Part ⅡThe study about the mechanism of gC1qR-induced apoptosis in cervical cancer cellsAims:We aim to investigate the effects of gC1qR expression on the mitochondrial function as well as the expression of pro-apoptotic protein p53 and explore the mutual relationships of gC1qR expression,mitochondria dysfunction,p53 and the apoptosis of cervical cancer cells to preliminarily clarify the potential molecular mechanism behind gC1qR inducing apoptosis.Methods:we constructed the vector gC1qR(Virus without gC1qR served as negative control)and applied DCFH-DA,Fluo-4AM,JC-1 to respectively detect mitochondria function.Besides,Western blot was used to evaluate the effects on p53.Next,on the basis of the construction of the gC1qR vector and empty vector,we respectively(1)cotransfected them with the empty pCB6,pCB6+p53wt,pCB6+p53mt at 72h;(2)cotransfected them with p53 siRNA,negative siRNA at 72h and applied WST,Transwell,Flow cytometry,DCFH-DA,Fluo-4AM,JC-1 to detect the changes of mitochondria function and the biological behaviours of cervical cancer cells.Results:We preliminary found that gC1qR could upregulate the levels of ROS,Ca2+,p53 and decrease mitochondria membrane potential in a time-dependent way.Furthermore,when we cotransfected gC1qR vector with wild-type p53,the levels of ROS,Ca2+,apoptosis,mitochondria membrane potential,viability,and migration increased or decreased significantly.However,the effects of gC1qR on mitochondria functions and the biological behaviours of cervical cancer cells can be abolished when gC1qR vector was cotransfected with mutant p53 or p53 siRNA.Conclusion:gC1qR could induce mitochondria dysfunction and increase the protein p53.Furthermore,gC1qR regulates apoptosis in cervical cancer cells via a p5 3-dependent mitochondria dysfunction pathway.