| Objective: By exploring the relationship between HR-HPV multiple infections and clinical features of cervical cancer,we analyzed the relationship between HR-HPV multiple infections and the malignancy degree of CC,which may provide the basis for clinical judgement of prognosis of cervical cancer with HR-HPV multiple infections.By exploring the mechanism of different clinical features caused by HR-HPV multiple infections,we may provide a new basis for clinical treatment of HPV.Methods: 1.From March 2011 to June 2015,we collected the paraffin section of 553 patients with a pathological diagnosis of cervical cancer from the First Hospital of Jilin University and the Second Hospital of Jilin University,all samples were paraffin embedded and stored at room temperature.DNA samples were taken from the tissue sections of the cancer foci after DNA extraction with a DNA extraction kit.HPV detection was done by use of PCR and genotyping by HPV Genotyping kit.Data analyses by Chi-square(χ2)tests using the SPSS statistical software package version 12.2.We chose 8 samples with HPV16 and HPV18 co-infected and designed the prime based on the standard sequence of HPV16 or HPV18 E6/E7,then paired the forward prime and reverse prime of HPV 16 and HPV 18 for Nested PCR to amplificated the sequence of E6/E7 as well as the possible recombination sequence.Connected the PCR products with p GEM-T vector and use genetic enzyme to cutting identification.Two-way gene sequencing of recombinant plasmids and analyzed by DANMAN.Results: 1.Among the 553 patients after surgery there were 547 HPV-positive women,430(78.6%)women were found with HR-HPV single infection and 117(21.4%)with multiple infections.The most prevalent HPV type still was HPV16 both in single infection(n=410 95.3%)and multiple infections(n=116 99%)and was significantly higher than it was reported before.HR-HPV multiple infections were higher in ASC(66.7%)than that in SCC(20.9%).The proportion of HR-HPV multiple infections(28.6%)in the poor differentiation group was significantly higher than that in the high/moderate differentiation group(19.1%)(P=0.005).HR-HPV multiple infections in HPV 18(81.3%)was significantly higher than that in other type of HPV infection.The proportion of multiple HPV-16 infections in the low differentiation group(29.9%)was significantly higher than that in the high/moderate differentiation group(19.5%,P = 0.032).The number of HPV-16 infection 491(96.7%)was higher than the other type study 17(3.3%).The proportion of multiple HPV-16 infections was higher in patients with ASC(75.0%)than those with SCC(21.7%,P = 0.006).The rate of HPV-18 infection in patients with ADC(18.5%)was significantly higher than that in patients with SCC(12.8%).2.There are point mutations of the E6/E7 gene in HPV16/HPV18 double-infected patients.By gene sequence alignment of the HPV E6/E7,at the 177 th base T,two samples were mutated to G,and 2 samples were mutated to A,which mutating the 32 nd Asp(GAT)of HPV 16E6 to Glu(GAA/GAG).Two samples of 647 base A were mutated to G,which mutated the 28 th Asn(AAT)of the HPV16 E7 protein to Ser(AGT).Besides,there are 5 samples at the 665 th base from G to A,which are nonsense mutations and all encode Glu(GAG/GAA).By gene sequence alignment of the HPV 18 E6/E7,in the 5 samples,484 th bases were mutated from C to T,position 548 th was mutated from A to C,and 750 th was mutated from C to T,548 th was mutated from A to C,and 750 th was mutated to C.All mutations were nonsense mutations,encoding Phe(TTC/TTT)and Arg(AGA/CGA)and Ala(GCT/GCC),respectively.Conclusions: 1.Our study provides important information that HR-HPV multiple infections can increase the malignant degree of cervical cancer.Different type of HPV may interact with each other and influence the clinical features of cervical cancer.2.There are point mutations of the E6/E7 gene in HPV16 in Jilin area,the function remained to be examed. |
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