Studies On Photodynamic Therapy For Cervical Cancer And The Clone And Construction Of Theraputic Antigen HPV18E6E7

Cervical cancer is the second leading cause of cancer death among womenworldwide. Statistics indicated that there were about500000new cases of cervicalcancer worldwide every year, of which200000died. Traditional treatments ofcervical cancer such as surgery, radiotherapy and chemotherapy have their ownlimitations, and can not cure cervical cancer. In recent years, tumor photodynamictherapy and tumor vaccine have been growing concerns. Therefore, in this study twosubjects were carried out according to the topic of cervical cancer treatment. One isthe research of photo-induced antitum or activities of hypocrellins to HeLa cells, theother is construction of therapeutic vaccines for cervical cancer with HPV18E6E7asantigen.Photodynamic therapy (PDT) is a minimally invasive therapeutic modalityapproved for the treatment of cancer diseases and many other non-oncologicaldisorders. In this study, a congener of Hypocrellins, EDAHB, was selected toinvestigate the HeLa cell death triggered by PDT. The result suggested that EDAHBexhibits a strong absorption shoulder peak at640nm and a high quantum yield ofboth1O2and O2-. It showed a much higher photopotentiation factor than HB. Thephototoxicity of EDAHB to HeLa cells occurred via a mitochondria/caspase apoptosispathway. This study showed EDAHB to be a promising candidate of photosensitizerfor anti-cervical tumor PDT.In the current work, the superior genetic coden of human gene were referred tooptimize and synthesize the full HPV18E6and E7using the molecular biologytechnology to enhance the gene’s expression in the mammilian cell. Meanwhilesite-directed mutageneses were introduced to eliminate the carcinogenicity of the E6and E7genes and the genes were fused to enhence the immunogenicity of the E6andE7proteins. Lastly, the recombinant plasmids of pDC315、pVR and pcDNA3.1(+)containing HPV18E6E7optimized fused wild/mutant genes were constructedsucessfully. The works laid the foundation for further construction of HPV18E6E7gene-based anti-cervical cancer vaccines.This work can provide a fundamental base for in-depth study of the developmentof theroputic vaccine.