Development Of Sensitive HPV Genotyping Method For Cervical Screening And Study Of HIF-1? And C-myc As Biomarkers For Cervical Cancer

Cervical cancer is the most lethal cancer for women.Fortunately,HPV has been found to be causative agents of over 99%of this cancer.Therefore,HPV screening plays an important role in primary cervical cancer screening.However,there are different HPV types which differ in their carcinogenic risk and areal distribution.This means that it would be more valuable to develop HPV genotyping methods which are still in lack for sensitive detection.On the other hand,HPV infection is not the only necessary factor for the progression to cancer.The change in host cells also contributes a lot to the cancerization.To study the change and find the key biomarkers,like genes and proteins,may be valuable not only for early diagnosis,but also for study of cancer therapy as well.In this study,we established a novel HPV genotyping method and explored potential biomarkers in the progression of cervical cancerous lesion.The results were promising for early diagnosis of cervical cancer and cancer therapy.In chapter one,an introduction on the background of HPV and cervical cancer was given firstly.Then,a review of HPV genotyping methods was made.The molecular mechanism of cervical cancer was described in detail,so that the potential biomarkers for therapy can be proposed and verified in the following.Finally,the idea and research program were put forward.In chapter two,we developed a new HPV genotyping methods based on PCR-RFLP-MCE system.The method was designed for 47 kinds of HPV types and compared with cytological test and DNA sequencing.The results turned to be 4-fold higher in HPV infection rate than cytological test.Two thirds of all positive samples were single infection.Eleven kind of HPV types were identified with good compatibility of over 90%.The PCR-RFLP-MCE automatic typing method showed reliability and for cervical cancer diagnosis.In chapter three,we primarily determine cooperation of HIF-la and c-myc in HPV 16 infectioned cervical cancer cells.Based on intramolecular DNA G-quadruplexes in promoter regions,we used G-quadruplex-interactive agents to study the gene expression of HIF-la,c-myc,p21 and VEGF.After stimulated by TMPyP4,two markers were inhibited as prediction,and G1 arrest can be observed.This result laid foundation for further study of biomarkers and new drug targets.At last,a summary was made for the whole work,as well as an outlook on expanding utility of PCR-RFLP-MCE genotyping method and the study of gene therapy.