Detection Of Human Papillomavirus And Its Study On Epidemiology Of Non-Small Cell Lung Cancer

Research Background and ObjectiveLung cancer is one of the highest morbidity and mortality malignant tumor in the world.Non-small Cell Lung Cancer is the most common type of lung cancer.Although it has been widely recognized that smoking and air pollution are the important risk factors of lung cancer,the exact etiology and pathogenesis are not very clear yet.HPV is an epitheliophilic spherical DNA virus.Persistent infection of HPV is the main cause of cervical cancer,but its role in lung cancer remains to be further studied.In this project,485 cases of lung cancer and 283 cases of adjacent normal tissues were detected by Touchdown PCR,and the infection rate of HPV between lung cancer group and paracancerous group were compared.The distribution of HPV genotype in lung cancer was observed by sequencing,so as to clarify the relationship between HPV infection and lung cancer.In addition,the distribution of HPV genotypes in cervical tissues in different regions,but it is not clear whether HPV genotypes have similar distribution characteristics in cervical tissues and lung cancer tissues in the same area.In this study,we compared the distribution of HPV genotypes in cervical tissued and lung cancer tissues in order to understand the differences and similarities of HPV genotypes in different tissues.The detection of HPV is widely used in the screening of cervical cancer.Detection of HPV is great significance for early diagnosis,treatment and prognosis evaluation of HPV infection.At present,clinical laboratories mainly use PCR and other methods based on molecular diagnostics to type and detect HPV.There are many commercial detection platforms and kits in the market,and different methods cover different types of HPV.The detection performance and clinical applicability of products need to be further studied.In this study,we give some advice about the selection of clinical HPV detection methods by comparing the detection ability of the method of hybridization,microfluidic and isothermal amplification.Methods1.A total of 768 formalin-fixed paraffin-embedded samples,of 485 lung cancer and 283 non-cancer controls were collected in the First Affiliated Hospital of Guangzhou Medical University.HPV DNA was detected and genotyped using Touchdown PCR and sequencing.The obtained sequences were compared with known Gen Bank sequences by BLAST tool to determine the HPV genotype.The infection rate of HPV in lung cancer patients with different clinicopathological features were analyzed.In addition,the distribution of HPV genotypes in cervical tissues and lung cancer tissues were analyzed and compared in this study.2.A total of 94 Samples of female cervical exfoliated cells were collected,and their HPV infection rate was detected by the methods of hybridization,microfluidic and isothermal amplification.The consistency of the results of each method was compared by calculating the Kappa value.A two-sided P-value of<0.05 was considered statistically significant.Result1.We found 23.92%(116/485)of lung cancer specimens were positive for HPV DNA,while 9.54%(27/283)of non-cancer controls were positive for HPV DNA(p<0.05).Within the group of age<60 years old had higher HPV infection rate than patients≥60 years old(p=0.033),and female had higher HPV infection rate than male(p=0.041).There was no significant difference in HPV infection rate among lung cancer patients with different histopathological types,smoking history and tumor stages.A total of 8 HPV genotypes of HPV3,6,11,16,18,58,59 and 81 were detected in lung cancer tissues.The detection frequency of HPV18 and 81 in lung cancer patients was significantly higher than that of non-cancer control patients.The infection rate of the top three genotypes were HPV81,HPV16 and HPV18,which account for 72.42%(84/116),12.07%(14/116),8.62%(10/116)of the positive number of lung cancer group,respectively.In the paracancerous group,the three HPV genotypes were HPV81,HPV16 and HPV59,which account for 85.19%(23/27),7.41%(2/27)and 3.70%(1/27)of the positive number.The HPV infection rate in cervical tissue was 22.95%(2208/9621),with HPV 52,16 and58 as most common.The detection rate of HPV16/18 in cervical tissue and lung cancer tissue was similar,but the detection rate of other genotypes was very different.However,HPV81 was the most common in low-risk HPV,both in cervical tissue and lung cancer tissue.2.The detection rates of HPV were hybridization 42.6%(40/94),microfluidic 38.3%(36/94),isothermal amplification 33%(31/94).The 17 samples were tested positive,and 50 samples were negative by the 3 methods,and the result consistency was 71.3%(67/94).After unification the detection range of each method,the detection results between any of the two methods has a excellent consistency(Kappa = 0.75,P < 0.05).Conclusion1.The infection rate of HPV in lung cancer group was higher than paracancerous group.We conclude that HPV18 and 81 were associated with lung cancer,however the infection of HPV16 was not correlated with lung cancer.The infection rate of HPV is related to age and gender of patients.The distribution of HPV genotypes in cervical and lung cancer samples in the same area was somewhat similar.2.The three methods have excellent consistency in the detection of HPV.This study provides the data reference for the choose of HPV detection methods.