Study On The Association Between HTERT And High-Risk HPV Infection In Cervical Lesions

Objective:To investigate the relationship between the human telomerase reverse transcriptase (hTERT) and high-risk human papillomavirus (HR-HPV) infections in cervical intraepithelial neoplasia(CIN) and cervix carcinoma.To explore the amplification of hTERT gene in cervical exfoliated cells by fluorescence in situ hybridization (FISH) on screening cervical precancerous lesions.To discuss the risk of HR-HPV infection predicted by hTERT serum mRNA levels by real-time fluorescent quantitative (qRT-PCR).Methods:1.The cervical exfoliated cells and serum specimens were collected from34cases of normal cervix,31cases of CIN grade Ⅰ,33cases of CIN grade Ⅱ,34cases of CIN grade Ⅲ,20cases of cervical carcinoma patients who were diagnosed by Second Hospital Affiliated to Tianjin Medical University Department of gynaecology colposcope directed biopsy histopathological examination.2.The polymerase chain reaction-reverse dotblot hybridization (PCR-RDB) for detection of HPV DNA positive cases.3.FISH technology to detect the amplification of hTERT gene.4.the level of serum hTERT mRNA expression was detected by qRT-PCR.5.hTERT maybe used in the diagnosis of cervical precancerous lesions and predict the risk of HR-HPV infection.It will provide more accurate, more reliable, more foresee index for CIN than TCT,HPV.Results:1.Twenty subtypes of HR-HPV were detected including HPV16,18,26,31,33,35,39,45,51,52,53,56,58,59,66,67,68,69,73and82. The incidence of HR-HPV infection was higher in CIN Ⅱ group (72.73%), CIN Ⅲ group (85.29%) and cervix carcinoma group (90.00%) than that of normal cervical epithelium group (20.59%). There was no significant difference in the positive rate of HR-HPV DNA between CIN Ⅰ group (54.84%) and normal cervical epithelium group (P<0.005).2.With the increase of the degree of cervical lesions, the positive rate of hTERT gene amplification increased gradually (P<0.05).The positive rate of hTERT gene amplification was higher in cervical cancer (80%) than that of the control group (0), CIN Ⅰ (3.22%), CIN Ⅱ (18.18%), but cervical cancer and CIN Ⅲ group (41.18%) was not statistically significant differences (P>0.005).3.The abnormal amplification of hTERT gene in cervical exfoliated cells were positive in serum of patients with mRNA, while hTERT gene without amplification were negative in serum of patients with mRNA.4.hTERT gene amplification, mRNA positive was positively correlated with HR-HPV infection (r=0.238, P<0.05). The level of hTERT mRNA in cervical cancer, CIN Ⅲ, CIN Ⅱ with HR-HPV imfection were higher than that of HR-HPV negative (P<0.05).5.With the increasing degree of cytological lesions, hTERT gene amplification and the mRNA positive rate gradually increased (P<0.05), cancer group (80%) was higher than that of NSIL (0), ASC (0), LSIL (2.7%), HSIL (28.6%), there were no significant differences between the NSIL and ASC group (P>0.005).6.Results of the three kinds of detection (TCT, HPV, hTERT):the specificity of hTERT were higher than those of HPV and TCT detection, it can make up the deficiency of HPV, TCT lower specificity.Conclusion:1.hTERT assay can be used as a tool for molecular screening of cervical cancer and precancerous lesions, also used in the diagnosis of early cervical lesions.2.The incidence of HR-HPV infection was positively correlated with hTERT gene amplification and expression in cervical lesions.HR-HPV infection may be early events before abnormal amplification of hTERT gene and expression, hTERT may predict the risk of HR-HPV infection.