Research On Molecular Epidemiology Of Human Papillomavirus In Guangdong Area

Cervical cancer is one of the most common cancers of female, and it is at thehead of female cancer in China. The epidemiology data demonstrates that HPVinfection is the main cause of cervical intraepithelial neoplasia and cervicalcancer.The HPV can be divided into different types according to its variation of itsnucleic acid array. Being unlike means that the similarity point of isogeny of the DNAof the virus and L1, E6 and E7(ORFs) is smaller than 90%; if the similarity point isbetween 90% to 98%, it is a subtype; if the similarity point is greater than 98%, it isvariants. It has been demonstrated the different HPV virus types have the differentpathopoiesia ability. We may divide subtype of HPV into the low-risk type andhigh-risk type according to its difference of pathopoiesia ability and the cancerogenicdanger. Infection rate of HPV is different in different area, and the type carried isdifferent. What’s more, it is different of the type related to cervical cancer. Therefore,knowing the popular situation and popular trend of HPV in Guangdong province willbe helpful to control the HPV infection in China.At present, HPV research mainly focuses on the research of polymorphism ofHLA, infection of HPV and cervical cancer, HPV epidemiology investigation andmolecule diagnose method research, research on mutant HPV separation appraisal,the molecule characteristic and pathopoiesia relation, HPV pathopoiesia mechanismresearch, and research on vaccine and prevention and cure technology. The homelandscholars concentrate on study the aetiology of the epidemiology of the sexuallytransmitted diseases. Meanwhile there are some research reports about the separating of an appraisal. But there are no relative research about the HPV molecule evolutionof the clinical diagnosis case in some high dangerous area and molecule epidemiology.As cervical cancer is a common female cancer, HPV infection has close relation ofthe development of cervical cancer. This research will mainly focus on the followingaspects from HPV virus.1. The HPV types of infection of human papillomavirus of women withcervical lesions in Guangdong areaAdopting PCR direct measure, we make investigations of 1400 patients withcervical lesion in Guangdong. Among them there are 1200 HC-Ⅱpositive specimensand 200 HC-Ⅱnegative ones. We divide the HPV gene in order to inquire into theHPV type and changing condition infected by Guangdong women. Study shows thatin the 1200 HPV DNA positive specimens there are 30 HPV genetypes(subtype),about 84.5%(1014/1200),single infection rate 93%(943/1014), 71commixture infection rate 7.0%(71/1014). The infection of each genotype and theirinfection rates in proper order are HPV16 (24.9%)、58 (17.4%)、33 (13.5%)、11(10.3%)、52 (6.9%)、6 (4.5%)、18 (3.3%)、53 (2.8%)、81(2.8%)、66 (2.4%)、31(2.4%)、39(2.0%)、84(2.0%)、56(1.6%)、54(0.8%)、59(0.8%); Andthere are 3 people of HPV CP83043, and only one is affected by HPV1、7、8、45、46、51、63、64、68、69、70、75、76 for eachtype, all together 16people, 1.7%. Inthe 200 HC-Ⅱnegative specimen there are 3 types of HPV genotype (sub type), 9samples, occupied 4.5% of masculine gender. The genotype and their infection inproper order is HPV 58 (4 samples, 44.4%), 16 (3 examples, 33.3%), 33 (2 examples,22.2%) and it has no commixture infection. And we discover the HPV46、52、81、84、CP8304 type exists in Guangdong area for the first time.This research discovers that there are at least 30 kinds of HPV subtype in thecells chipped off from the neck of uterus in Guangdong area. Among them, there are16 kinds HPV of high-risk types(16,58,33,52,18,53,66,31,39,56,54, 59,45,51,68,69), and HPV16,58,33,52,18 is the usual ones. Atthe same timewe have found 14 kinds of HPV of low risk (11, 6,81, 84, CP8304, 1, 7,8,46, 63,64,70,75,76). HC-Ⅱnegative specimen can not remove HPV infection, and itmust be judged by clinical pathology.2. Research on the subtypes and infection of HPV of patients with cervicalcancer and cervical intraepithelial neoplasia within Guangdong areaAdopting the PCR direct measure, we carded out the HPV gene sequence of thedifferent cancer organization cell specimen of cervical cancer to divide the differenttypes in order to inquiring into the HPV types and variation of the cervical cancer ofwoman in Guangdong area. Research on the HPV type and their influencing factor ofwomen cervical cancer patients in Guangdong will provide experiment basis fordeveloping HPV precautions. Study result points out that the HPV infection rate ofcervical lesion of patient in Guangdong area is 54.6%;with the cervical cancer,HPV16, 33, 18, 58 accounting for 31.78%、23.36%、16.82% and 3.74%respectively.This study shows that age between 21 and 40 is the high time of HPV infectionof all kinds of cervical lesion while the high time of the cervical cancer is between 31and 50. The high-risk types of HPV infection rate of cervical cancer of different agegroup does not have notable difference. But the rate of the age between 31 and 40 andage between 41 and 50 will be the higher ones, with 85.2% and 82.5%. Results alsoshow that the morbidity age is different in different times. The morbidity age between1975-1984 and 1985-1994 does not have difference (P=0.970), but it is differentfrom 1995-2004 (P=0.005). It shows that cervical cancer age is over 10 years ahead.That is to say, the high-risk types infection of combined neck of uterus will becomecervical cancer in 10 years. And in the recent 10 years the cervical cancer happened10 years earlier. A prophylactic vaccine to prevent infection with 1 or more ofhigh-risk types has the potential to substantially reduce the incidence of cervicalcancer and its precursor lesions in 21-40 ages.3. The genetic polymorphism analysis of HPV L1 of cervical lesionorganization in Guangdong areaThe analysis of the polymorphism of HPV L1 gene helps the diagnose analysisand epidemiology analysis clinically, also has importance in genetic engineering prophylactic vaccine. This research analyzes the pathological changes of cervicallesion in Guangdong, especially analyzes the array of the high-risk and high-riskHPV L1 gene, to inquire into the polymorphism of HPV L1 gene infected the womenin Guangdong area. Research shows that the polymorphism of the low-risk HPV L1gene in Guangdong area is: To understand the molecular evolution of the HPV inGuangdong, the sequences were compared with representative HPV sequencesobtained from Genbank. The phylogenic analysis revealed that HPV 33、52、58 inother areas originated from ones in Guangdong. The HPV16 L1 gene has manymutation, the hot mutations are M368K, S375F and D412E. The local mutant plantcoexists with the wild plant and gives first place to the local mutant plant. There ispossibility that the HPV 16 viruses L1 genes abroad come from Guangdong area. Butthe mutation of C350W, R364S and T366N may be the characteristic of thepolymorphism of HPV58 L1 gene in Guangdong area. By evolution analysis,TheHPV efficacious prophylactic vaccine in other areas will be nonefficacious inprevention of incident and persistent cervical infections with HPV in Guangdong, andassociated cytological abnormalities and lesions. Vaccination in Guangdong areaagainst such infections could substantially reduce incidence of cervical cancer. HPV6L1 H315N、E431Q variants, HPV16 L1 M368K、S375F、D412E variants, HPV58L1C350W、R364S、T366N variants and genetic polymorphism of HPV18、33、39、45、52、53、54、58 will be thought over before new prophylactic vaccine suitablefor women in Guangdong are developed.4. The genetic polymorphism analysis of high risk HPV (16,33,58) E6 andE7 of cercival lesions organization in Guangdong areaE6 and E7 in the transcribing area in the early phase are mainly connected withthe conversion function of virus cell and carcinogenicity. And it has the anti-formactivating activity to the transcription of virus gene and cell gene. This study analyzesthe pathological changes of cervical lesion in Guangdong province, especially, thearray of the high risk HPV E6, E7 gene in cervical cancer organization to inquire intothe polymorphism of HPVE6 and E7 gene infected by women in Guangdong. Theresult points out that the most common genetic mutations of HPV16 E6/E7 in Guangdong area are E6 D32E and E7 N29S. We guess that there are E7 N29S and E6D32E of HPV16 in Guangdong area. Besides, there are some mutant ones carried bythe movement of population. The hot mutations of HPV33 E6/E7 are: E6 K35N, E6K93N are consistent with what reports in the homeland and Japan; E7 A45E andA50P; HPV33 E6 and E7 genes in Guangdong area are concomitance of mutant plantand wild plant. The hot mutations of HPV58 E6/E7 are: E6 K93N, E7 T20I and G63Sare consistent with HPV58 dissociation in Hong Kong; E7 T20I is near theL-Xaa-C-Xaa-E area and is related to the adjustment of p107 and p130;E7 G63Smakes the serine phosphorylated and the conformation and the polarity of proteinmay change, therefore the biology function changes; At the same time, there are someother dissociation in E6/E7, but some of them are consistent with the standard one.From the evolution tree we can see that there are local mutant plant, wild plant anddissociation of other places of HPV58 E6 and E7 in Guangdong area. HPV16、HPV33、HPV58 E6/E7 variants will be thought over before new therapeutic vaccinesuitable for women in Guangdong are developed.From this research, we discover for the first time that there are more than 30HPV subtypes in the cells chipped off from neck of uterus (HC-Ⅱpositive specimens)in Guangdong area. And there are 15 of high risk, among them HPV16, 58, 33, 52and 18 are the common ones. HC-Ⅱnegative specimens can not remove the HPVinfection. This study shows that age between 21 and 40 is the high time of HPVinfection of all kinds of cervical lesion while the high time of the cervical cancer isbetween 31 and 50. The high risk HPV infection of combined neck of uterus willbecome cervical cancer in 10 years. And in the recent years the cervical cancerhappened 10 years earlier. HPV16, 33 and 18 are a main cervical cancer pathopoiesiatype, with the certain crowd or region characteristic. The polymorphism of high riskHPV L1 gene in Guangdong area is: The HPV16 L1 gene has many mutations,E431Q being same with one internal virus plant and a European one. The HPV11 L1gene is relatively conservative. And the HPV46、52、81、84、CP8304 subtypes firstdiscovered exists in Guangdong area for the first time. The polymorphism of high risk HPV L1 gene in Guangdong area is: The HPV16 L1 gene has many mutation:M368K,S375F and D412E. The hot mutation of HPV58 is T375N/I412V/N422D. The hotmutations of HPV33 E6/E7 are: E6 K35N, E6 K93N. The Asia type (E7 N29S)coexist with the local mutant plant (E6 D32E) and other mutant plants. The hotmutations of HPV33 E6/E7 are E6 K35N and E6 K93N. HPV33 E6 and E7 genes inGuangdong area are concomitance of mutant plant and wild plant. The hot mutationsof HPV58 E6/E7 are E6 K93N, E7 T20I and G63S; E7 T20I is near theL-Xaa-C-Xaa-E area; E7 G63S makes the serine phosphorylated and the biologyfunction changes; From the evolution tree we can see that there are local mutant plant,wild plant and dissociation of other places of HPV58 E6 and E7 in Guangdong area.HPV 6L1 H315N, E431Q variants, HPV16 L1 M368K、S375F、D412E variants,HPV58L1C350W、R364S、T366N variants and genetic polymorphism of HPV18、33、39、45、52、53、54、58 will be thought over before new prophylactic vaccinesuitable for women in Guangdong are developed.HPV16、HPV33、HPV58 E6/E7variants will be thought over before new therapeutic vaccine suitable for women inGuangdong are developed.