Analysis Of Human Papilloma Virus Infection And Study On The Mechanism Of HVJ-E Induced Apoptosis In Hela Cells

Human papillomaviruses(HPV)spread among people widely and infect the skin and mucous membrane to cause benign or malignant lesions.The study find that verruca vulgaris,condyloma acuminatum,cervical cancer,skin cancer,vaginal cancer,penile cancer,anal cancer,breast cancer and so on are all associated with HPV infection and 95%of cervical cancer is caused by high risk HPV.The infection of HPV has age and regional differences,which infection rate and subtypes of HPV are different.We analyze the conditions of HPV infection in 10473 women and have a preliminary understanding of the prevalence of HPV infection among women in Yangzhou area,to provide evidence for the prevention and treatment of HPV infection.In order to explore a new method for tumor therapy,we study on the mechanism of HVJ-E induced apoptosis of HPV 18 positive Hela cells.Part one Analysis of human papillomavirus infectionObjective:To uderstand the situation of HPV infection among women in Yangzhou and to provide evidence for the prevention and treatment of HPV infection.Methods:Collecting cervical exfoliated epithelial cells from 10473 female outpatients aged 18-86 in dermatology and gynecology clinic of Subei People's Hospital from October 2015 to October 2016,the subjects were divided into?29,30-39,40-49,50-59,?60 five age groups,we used PCR-reverse dot blot hybridization method to detect 21 gene subtypes of HPV,then we analyzed distribution characteristics of HPV subtypes and used chi-square to compare infection rate.Results:10473 cases of female patients,2577 cases are positive,the infection rate is 24.61%.Single HPV infection are 1819 cases and multiple infection are 758 cases.The infection rates and positive constituent ratio of five age groups are 23.88%,23.36%,23.77%,28.25%,32.27%and 17.66%,27.40%,34.65%,14.82%,5.47%,respectively.The detection rate of Hr-HPV infection is 28.4%,of which HPV-16,52,58,53,39,51 are in the top six and their detection rate are 4.54%,4.33%,3.64%,2.96%,2.17%,1.96%,successively and the detection rate of Hr-HPV18 is low,only 1.46%.The detection rate of Lr-HPV infection is 5.99%,among which HPV-81,11,6 and 44 are in the top four,and their detection rate are 3.16%,0.93%,0.91%,respectively.Conclusions:The distribution of HPV infection has age and region difference in Yangzhou area;the peak population of HPV infection is 30-49 years old,the most common high risk and low risk HPV subtypes are16,52,58,53,39,51 and 81,11,6,44,respectively.The detection of HPV 21 subtypes have instructional significance for the prevention and treatment of HPV infection in Yangzhou area.Part two Study on the mechanism of HVJ-E induced apoptosis in Hela cellsObjective:We choose HPV 18 positive Hela cells as the research object to explore the mechanism of HVJ-E induced apoptosis in Hela cells.Methods:Hela cells were treated with different concentrations of HVJ-E,the cells morphology were observed by inverted microscopewe,the cells viability and apoptosis rate were detected by CCK-8 and FACS assay.We assayed p53,Bcl-2/Bax,Caspase and MAPK signaling pathway proteins by Western blot.To confirm Caspase and MAPK signaling pathway play an important role,specific inhibitors,Z-VAD-FMK(50?M),U0126(18?M),SP600125(5?M)and SB203580(12.5?M),targeting Caspase,ERK,JNK and p38,respectively,were added to cells 20min,2h,40min,2h prior to HVJ-E.We used the method of CCK-8 and FACS to detecte cells viability apoptosis rate.And we assayed Caspase-3,PARP,p-ERK1/2,p-JNK,p-p38 proteins by Western blot.Results:HVJ-E causes Hela cells growth arrest,shrinkage and rupture.With the increase of HVJ-E concentration,Hela cells viability decreased,apoptosis rate increased,the expression of p53,Caspase-9,Caspase-3 and PARP protein increased and the anti apoptotic protein of Bcl-2 decreased,the apoptosis protein of Bax do not change.The ratio of Bcl-2 and Bax decreased with the increase of virus concentration.In the MAPK signaling pathway the expression of phosphorylated of ERK1/2,JNK and p38 is dose dependent.The inhibitor can down regulate HVJ-E induced apoptosis,reduce cell death induced by HVJ-E,and inhibit the expression of Caspase-3,PARP,p-ERK1/2,p-JNK,and p-p38 protein.Conclusions:HVJ-E can induce apoptosis of Hela cells and p53,Bcl-2/Bax,Caspase and MAPK signaling pathway play an important role in HVJ-E induced apoptosis of Hela cells.