Study On The Effect Of TLR3and HPV16E6Mutation In Cervical Lesions

ObjectiveTo explore the role of TLR3and high-risk type human papillomavirus (HR-HPV) load in progression of cervical lesions. To determine the association between TLR3and regulaition of HPV infection. To investigate distributional characteristics of mutations of HPV16E6oncogene in the patients with cervical lesions in this hospital and to explore the association between of HPV16E6variants and progression of cervical lesions.MethodsCervical exfoliated cells from54cases with positive HPV16were subjected to regular DNA extraction procedure. We design corresponding primers to amplify TLR3and HPV16E6inorder to determine them semi-quantitatively by PCR. The products of HPV16E6purified and sequenced, then were carried out blast analysis against standard strain in GenBank to evaluate the genetic mutaition.Results1. We splited54patients with different stages of cervical lesions into three groups, group A (including14chronic cervicitis,4CIN Ⅰ) has18cases, group B (including12CIN II-III,6cervical carcinoma in situ) has18cases, group C (invasive cervical cancer) has18cases. The ages between3groups have no significant differences (p=0.506).2. The expression of TLR3in3groups reduced gradually and the difference was significant (p<0.001). The expression of TLR3was (0.6676±0.0226),(0.6484±0.0353) and (0.5652±0.0330) respectively in group A, B and C. It was significantly higher in group A and B than in group C and the differences were both significant (p<0.001;p<0.001). The expression of TLR3in group A was little higher than group B and the difference was not significant (p=0.066).3. The expression of HPV16E6in3groups raised gradually and the difference was significant (p<0.001). The expression of HPV16E6was (0.3771±0.0266),(0.6600±0.0401) and (0.7393±0.0271) respectively in group A, B and C. It was significantly higher in group B and C than group A and the differences were both significant (p<0.001;p<0.001). The expression of HPV16E6in group C was much higher than group B and the difference was significant (p<0.001).4. Among the53cases sequenced successfully,4mutation spots were identified totally in26cases (49.06%). The most frequent sequence variation of HPV E6was T178G (D25E,43.40%), which is the same with type AS. The distribution of this sequence variation in group A (3/17) is lower than that in group B (9/18) and group C (11/18), the differences were significant (p<0.05;p<0.025). The distribution of this sequence variation was not significantly different between group B and group C (p>0.25).2cases in groupB and C occurred sequence variation T241G (3.77%). There are other two only one case gene mutations in group C, G188T (L30I,1.89%) and G543A(L147F,1.89%).5. The expression of TLR3and HPV16load was Negative correlated in cervical lesions (r=-0.637;p<0.001).Conclusions1. TLR3may play an important role in the regulation of HPV infection, it may be the main Immune response mechanism of the host against infected HPV. There may be significant correlation between abnormal expression of TLR3and progression of cervical lesions.2. HPV (especially HR-HPV) load has significant correlation with progression of cervical lesions; the mutation of HPV16E6may be important risk factors during the progression of cervical lesions.3. TLR3may play an important role in the regulation of expression of HPV16E6.