Research On High-risk HPV Genotype Test, Bypass And HPV16 Variants In Cervical Cancer And Precancerous Lesions

Objective: The main purpose of the passage is to evaluate the feasibility and application value of high-risk HPV genotype test during the cervical cancer screening, The expression and clinical significance of P16/Ki-67 cytological double staining in HPV positive and exfoliative cells of various cervical lesions; To study the pedigree distribution of infected HPV16 variants in Xinjiang area, as well as E6, E7 and LCR gene mutation; and to analyze the effects of HPV16 variants in the occurrence and development of cervical cancer and precancerous lesions. Methods: HR-HPV genotype tests were carried out for cervical cell samples in 564 female patients who were performed the cervical cancer screening as well as HC2 and TCT test with pathological results in the wards and outpatient of Peo Ple's Hospital of Xinjiang Uygur Autonomous Region between September 2014 and August 2015 using high-risk HPV genotype test system. There were 59 cases TCT smears used for detection of expression of P16 and Ki-67 proteins in cervical cytological specimens by using immunocytochemical method. Genomic DNA was extracted in patients with HPV16 positive cervical cancer and precancerous lesions. HPV16 DNA E6 and E7 genes and nucleotide fragments of LCR region were carried out the forward and reverse sequencing using PCR amplification. HPV16 pedigree distribution was confirmed via analyzing and comparing the HPV16 gene sequencing to analyze the mutation sites of nucleotide. Results: In the diagnosis of CIN2+, high-risk HPV genotypes is better than HC2, TCT and Hybr Miax genotyping detection.The difference has statistical significance(P<0.05). HR-HPV infection rate is varying in different TCT classification and tissue pathology grading. The difference has statistical significance(P<0.05). Along with the grade of cytology and pathological diagnosis rising, HR-HPV infection rate(P<0.05) and HPV16 infection rate(P<0.05) are increased respectively. All levels of cervical lesions are mainly based on HPV16 or with HPV16 infection. HPV16 is distributed in Uyghur and Han women without statistical difference(P>0.05). High-risk HPV constituent ratio in various age brackets has no statistical difference(P>0.05). High-risk HPV genotype test to ASCUS bypass is superior to HC2. P16/Ki-67 cytological double staining test has a high sensitivity(92%) and total coincidence rate(78%) in diagnosis of CIN2+. P16/Ki-67 cytological double staining test has a high coincidence rate(Kappa=0.569, P<0.05) with histopathological results. When having P16/Ki-67 cytological double staining test on ASCUS/ASC-H, it is more easily to find out the high-grade lesions(2? =16.327P<0.05). The positive rate of P16/Ki-67 cytological double staining in various cervical lesions: inflammation 39.13%(9/23), CIN1 20%(2/10), CIN2-3 91.30%(21/23), cervical cancer 100%(2/2). The difference has statistical significance(P<0.05). P16/Ki-67 cytological double staining rate shows an ascending trend along with aggravated lesion degree. E6 gene mutation rate is 80.00%(92/115) and the main mutation sites are T350G(59.78%) and T178G(18.47%). E7 gene mutation rate is 54.78%(63/115) and the main mutation sites are A647G(33.33%) and T846C(26.98%). LCR mutation rate is 23.48%(27/115) and the main mutation sites are C24T(74.07%) and C13T(25.92%). Compared the Uyghur women with Han women, Uyghur T350 G mutation rate is significantly higher than the Han. Han A647 G, T846 C and C24 T mutation rate is significantly higher than Uyghur. The diffidence has statistical significance(P<0.05). T350 G mutation in the Uyghur cervical cancer group is significantly higher than that of inflammation group(P<0.05). Moreover, the severity of mutation goes up along with lesions. Han T350 G, A647 G, T846 C and C24 T mutation rates in the inflammation group and cervical lesions group are significantly higher than cervical cancer(P<0.05). Uyghur C24 T mutation rate in the inflammation group is significantly higher than cervical cancer group(P<0.05). The difference has statistical significance(P<0.05). Infected HPV16 in Xinjiang Han group is mainly Asian type while infected HPV16 in Xinjiang Uyghur group is mainly European type. Conclusion: High-risk HPV infection is a main pathogenic factor of precancerous change and cervical cancer. All levels of cervical lesions are mainly based on HPV16 or with HPV16 infection. HR-HPV infection rate goes up along with increased cytology and pathological diagnosis grade. High-risk HPV genotype test to ASCUS diagnosis is superior to HC2. P16/Ki-67 cytology double staining positive rate is on the rise along with cervical lesions. P16/Ki-67 cytology double testing can effectively diagnose and bypass for ASCUS and HPV positive patients. HPV16E6 and E7 mutation may be related to cervical lesions. T350 G mutation may be the cause of the high incidence of cervical cancer in Uygur. Infected HPV16 is mainly Asian type among Xinjiang Han People while infected HPV16 is mainly Europe type among Xinjiang Uyghur people.