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Objective To detecte the differential metabolites and related pathways in Siha cells of cervical cancer by screening the inhibition of HPV16 E6/E7 expression based on 1H NMR metabolomics so as to identify the key metabolic markers involved in the development of high-risk HPV16 cervical cancer.Methods Siha cells were transfected with RNAi fragments to down-regulate the expression of E6/E7,which were divided into the normal control group(Siha cells),no-load group(si-NON),si-E6 group and si-E7 group,and their transfection efficiency was verified.1H NMR metabolomics was used to reveal the differential metabolites involved in interfering E6/E7 expression in Siha cells.Combined with MetaboAnalyst 5.0 online software,differential metabolites and related metabolic pathways were obtained.Results Fluorescence was observed by inverted fluorescence microscope.Western blotting results showed that compared with Siha group,the expression of E6/E7 in si-E6 group and si-E7 group was decreased(F=145.8,P<0.001).After down-regulating the expression of E6/E7,13 common differential metabolites,including Isoleucine,Leucine and valine,were detected.The results of MetaboAnalyst 5.0 online software analysis suggested that the above metabolites were mainly involved in the biochemical synthesis pathway of aminoacyl-trNA,biochemical synthesis pathway of isoleucine,Leucine and valine;There were 10 metabolic pathways of tyrosine,phenylalanine and tryptophan biochemical synthesis.Conclusion After HPV16 infection,changes of glucose and amino acid metabolism can promote the progression of cervical cancer,which provide a theoretical basis for the prevention and treatment of cervical cancer.
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@#Objective To investigate the value of human papilloma virus(HPV)DNA,HPV E6/E7 protein and thin-prep cytology test(TCT)in cervical intraepithelial neoplasia(CIN)and cervical cancer screening.Methods A total of 190 adult women who received early cervical cancer screening in Department of Gynecology,Zhuji People's Hospital from July 2021 to June 2022 were selected as the study objects.HPV DNA,HPV E6/E7 protein and TCT were detected,respectively,and further colposcopic biopsy was performed.The diagnostic efficacy of HPV DNA,HPV E6/E7 protein and TCT in patients with different lesions were compared.Results The positive rates of HPV DNA,HPV E6/E7 protein,TCT and combined detection in CIN3 and cervical cancer patients were significantly higher than those in cervicitis patients(P<0.05).The positive rates of HPV DNA,HPV E6/E7 protein,TCT and combined detection in cervical cancer patients were significantly higher than those in CIN1 patients(P<0.05).The positive rates of HPV DNA,HPV E6/E7 protein,TCT and combined detection in CIN2+ patients were significantly higher than those in CIN1-patients.The sensitivity,specificity,positive predictive value and negative predictive value of HPV DNA,HPV E6/E7 protein and TCT in the diagnosis of high-grade lesions were 90.80%,30.10%,52.32%and 79.48%,respectively.Conclusion HPV DNA,HPV E6/E7 protein and TCT can be used as screening methods for cervical cancer and precancerous lesions,with the combination of the three being the most sensitive.
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Purpose/Significance To explore the characteristics and clinical significance of differentially expressed genes closely re-lated to HPV E6/E7 by using bioinformatics.Method/Process The cervical tissue and clinical information of cervical cancer in TCGA and GTEx of UCSC are used as the training set.The expression profile chip GSE63514 related to cervical cancer in GEO is used as the validation set.Firstly,the limma package of R software is used to screen DEGs of tumor and normal samples,and Venn map of genes re-lated to E6/E7 protein in MigDB is made.Survival analysis is performed by survival kit and verified by ROC and protein expression lev-els.Secondly,key genes are obtained by copy number variation and methylation correlation.Finally,the specific co-expression network is constructed and enrichment analysis and immune infiltration analysis are performed.Result/Conclusion There are 101 differentially expressed genes related to HPV E6/E7 protein,and three genes are found to have significance after screening,namely E2F1,MCM4 and PCNA.At the same time,it is found that the genes in the specific coexpression network are significantly enriched in the DNA replication and chromosome organization pathways.Immune correlation analysis shows that key genes are significantly associated with CD4 T cells,B cells and neutrophils.DNA replication,chromosome organization,etc.,are the molecular mechanisms and key genes significantly related to the development of cervical squamous cell carcinoma and HPV E6/E7 encoded proteins.
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Objective To explore the clinical value of HPV E6/E7 mRNA in screening lesions at grade CIN2 and above.Methods A total of 120 cases with CIN and suspected cervical cancer treated in our hospital from January 2014 to September 2016 were selected.According to the results of pathological examination,60 patients with CIN2,CIN3 and invasive cancer were selected as the research group.60 patients with normal CIN1 and CIN1 were selected as control group.The results of TCT,HPV DNA and HPV E6/E7 mRNA were detected and analyzed.Results In 120 patients,the total positive rate of TCT was 55%,the total positive rate of HPV DNA was 87.5%,the total positive rate of HPV E6/E7 mRNA 60.8% HPV E6/E7;control group mRNA positive rate of 78.3% was significantly higher than the positive rate of HPV DNA 36.7%(P<0.05),HPV in the study group were E6/E7 mRNA positive rate of 85.0% was significantly lower than the positive rate of HPV DNA(96.7%);HPV E6/E7 mRNA CIN1,quantitative CIN2,CIN3 and invasive carci-noma were(4 867.31 ± 694.84),(8 943.51 ± 986.23),(28 243.10 ± 10 963.21),(3 610.84 ± 412.64)copies/mL;HPV E6/E7 mRNA quantitative CIN3 values were significantly higher than that of CIN 2,CIN1 and infil-tration of cancer and the control group(P<0.05),E6/E7 mRNA CIN2 quantitative HPV the value of CIN1 was significantly higher than that of control group(P< 0.05).Two methods of detecting HPV DNA and HPV E6/E7 mRNA in the screening of CIN2 and above lesions,the sensitivity of 84.5% DNA 93.8% HPV sensitivity was higher than that of HPV E6/E7 mRNA(P<0.05)HPV E6/E7 21.6%;specificity the speci-ficity of mRNA was 53.6% higher than that of HPV DNA(P<0.05),HPV DNA.HPV ROC curve E6/E7 mRNA were 0.581 and 0.681,the difference was statistically significant(P<0.05).Conclusion The specific-ity and accuracy of HPV,E6/E7 and mRNA in detecting cervical lesions over CIN2 and above are higher than those of HPV and DNA.It is of certain diagnostic value for screening cervical lesions over CIN 2 and above.
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Objective To evaluate the effect of combined detection of cervical E 6/E7 and liquid based cytology(TCT)in the screening of cervical cancer.Methods A total of 206 cases of high-risk patients with cervical cancer from March 2015 to March 2017 in Yongding Hospital of Suzhou were taken the cervical E6/E7 detection and TCT two screening methods respectively,and the pathological diagnosis was taken as the gold standard,the efficiency difference among TCT,cervical E6/E7 detection,and the com-bined detection of cervical cancer was compared.Results The sensitivity and negative predictive value of E6/E7 were significantly higher than those of TCT(P<0.05).The specificity of TCT was significantly higher than that of cervical E 6/E7(P<0.05).The accuracy of combined diagnosis was significantly higher than that of cervical E6/E7 and TCT alone.The difference was statistically significant(P<0.05).Compared with the cervical E6/E7,the specificity and positive predictive value of the combined diagnosis were higher,and the difference was statistically significant(P<0.05).The sensitivity and negative predictive value of the combined diagnosis were significantly higher than those of TCT,and the difference was statistically significant(P<0.05).The receiver oper-ating characteristic curve(ROC curve)showed that the area under the combined diagnostic curve(AUC)was significantly higher than that of the cervical E6/E7 and TCT alone,and the difference was statistically significant(P<0.05).Conclusion Cervical E6/E7 detection has a low specificity,and sensitivity of TCT is low;the combined detection of cervical E6/E7 detection and TCT has a high screening accuracy,and can improve the sensitivity and specificity of individual diagnosis.
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Objective To analyze the differences of positive detection rate and copy number of human papillomavirus (HPV) DNA and E6/E7 mRNA between different grades of cervical lesions, and evaluate their clinical values in early screen?ing of cervical cancer. Methods The cervical exfoliated cell samples from 154 women undergoing biopsy examination and 32 objects undergoing hysterectomy (control group) were collected in Tianjin Central Hospital of Gynecology Obstetrics in 2014. According to the pathological results of cervical biopsy, 154 samples were divided into low-grade squamous intraepi?thelial lesion group (LSIL, n=51), high-grade squamous intraepithelial lesion group (HSIL, n=71), and squamous cell carci?noma group (SCC, n=32). HPV DNA was tested with hybrid capture technology, and E6/E7 mRNA was detected with fluores?cence quantitative hybridization. Immunohistochemistry was performed by detecting E6/E7 protein in all patients after sur?gery or cervical biopsy. Results Combined results of HPV DNA and E6/E7 mRNA demonstrated that the positive detection rate was significantly lower in control group than that of all levels of lesion groups (P 10 000 E6/E7 were significantly increased in high-grade squamous intraepithelial lesion group. Immunohistochemical results showed that the positive detection rate of E6/E7 was significantly lower in control group than that of all levels of lesion groups (P<0.05). The positive rate of E6/E7 was significantly higher in the high-grade squa?mous intraepithelial lesion group than that of low-grade group (P<0.05). Conclusion HPV infection is closely related to cervical abnormalities, which is one of effective measures for early screening of cervical cancer. The negative result of HPV DNA is very helpful to exclude the cervical abnormality, whereas the positive detection of mRNA has great value in predict?ing the disease. Combined results of positive detection and copy number make a comprehensive evaluation for the risk of cer?vical lesions.
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Objective To investigate the value of HPV E6/E7 mRNA test combining liquid-based cytology test in cervical cancer screening. Methods A total of 377 samples from Wenzhou People's Hospital from June 2014 to September 2015 were collected and screened by HPV E6/E7 mRNA test combining with liquid-based cytology test , and the results was compared with the findings from the gold criteria of histology and pathology. Results The combination of HPV E6/E7 mRNA test and liquid-based cytology test can enhance the testing sensitivity and specificity. The sensitivity of the combination of HPV E6/E7 mRNA test and liquid-based cytology test for the diagnosis of LSIL was 94.41%, and that for the diagnosis of HSIL was 96.36%. Based on the gold criteria of histology and pathology , the sensitivity , specificity , positive-predictive value and negative predictive value of HPV E6/E7 mRNA test for the diagnosis of HSIL was 90%, 60.67%, 48.53% and 92.49%respectively. The sensitivity, specificity, positive-predictive value and negative- predictive value of liquid-based cytology test for the diagnosis of HSIL was 72.73%, 75.28%, 54.79% and 87.01% respectively. Conclusions The sensitivity of HPV E6/E7 mRNA test is superior to that of liquid-based cytology test , while the specificity of HPV E6/E7 mRNA test is inferior to that of liquid-based cytology test. The negative predictive value of HPV E6/E7 mRNA test is more meaningful than that of liquid-based cytology test. The combination of HPV E6/E7 mRNA test and liquid-based cytology test can enhance the testing sensitivity , but it does not increase the specificity.
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Objective To explore the clinical value of examining HPV E6/E7 mRNA level in assessing cervical le?sions infected with high-risk human papillomavirus (HR-HPV). Methods The cervical epithelial cells were collected from 265 patients with HR-HPV infection, including 100 cases of neoplasia free/inflammation group (control group), 88 cas?es of cervical intraepithelial neoplasia (CIN)Ⅰ, 33 cases of CINⅡ, 28 cases of CINⅢand 16 cases of cervical carcinoma and the transcription of HPV E6/E7 mRNA level was examined using branched DNA (b-DNA) technology. Results The positive rate HPV E6/E7 mRNA were higher in CIN Ⅱ(81.82%), CINⅢ(89.29%) and cervical cancer group (100.00%) than tthat in control group (20.00%) and CINⅠ(35.23%) with significant difference, and there were no significant differences between other groups;The positive rate and transcription level of HPV E6/E7 mRNA in HSIL (high grade squamous intraepi?thelial lesion)and cancer group were significantly higher than normal, ASC(atypical squamous cell carcinoma) and LSIL(low grade squamous intraepithelial lesion) group (P<0.05). Conclusion The transcription level of HPV E6/E7 mRNA may re?flect the activity of the virus and the progression of disease, and could be use as an effective indicator to screen high grade cervical pathological changes and a complementary method of cervical lesion screening.