ABSTRACT
Objective To explore the diagnostic value of carcino embryonic antigen (CEA),squamous cell carcinoma antigen(SCC),human papilloma virus-E7 (HPV-E7) in cervical carcinoma.Methods A total of 107 cases of women patients treated in hospital from July 2013 to July 2015 accorded to the pathological examination results were divided into cervical cancer group 60 cases and CIN group 47 cases,another 50 cases of healthy people were selected as control group, and serum expression levels of HPV-E7, CEA and SCC in the three groups were detected by enzyme-linked immunosorbent assay.Results The serum HPV-E7, CEA and SCC in cervical cancer group were significantly higher than those in CIN group and control group (P<0.05).There was no significant difference in serum HPV-E7, CEA and SCC between CIN group and control group.The levels of serum HPV-E7, CEA and SCC in stage I-II were significantly lower than those of stage III-IV in patients with cervical cancer, and the difference with statistically significant between two groups(P<0.05).The area under the ROC of HPV-E7 was significantly higher than that of CEA and SCC (Z=2.914,2.951, P<0.05), and there was no significant difference in the area under the ROC between CEA and SCC (Z=1.580,P=0.057).Conclusion The serum HPV-E7, CEA and SCC in cervical cancer patients are significantly higher, the diagnosis value of HPV-E7 is higher which is expected to become one of the effective indicators of cervical cancer diagnosis.
ABSTRACT
The persistence infection of low-risk type (type 6 or type 11) of human papillomavirus (HPV) is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent is certain to be of value. The aim of this study was to achieve targeted inactivation of viral E 7 gene in keratinocytes using the reprogrammed clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) 9 system. To accomplish this, a universal CRISPR-Cas9 system for targeting both HPV6/11 E 7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E 7-transfromed keratinocytes, the expression level of E 7 protein was measured using western-blot analysis and the sequence of the E 7 gene was determined using Sanger sequencing. Cell proliferation was analyzed by CCK-8 assay, and cell apoptosis was evaluated by Hoechst 33258 staining, flow cytometry analysis and ELISA assay. The results indicated that both HPV6/11 E 7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E 7 led to inhibition of cell proliferation and induction of apoptosis in E 7-transfromed keratinocytes but not in normal keratinocytes. Our data suggested that the reprogrammed CRISPR-Cas9 system has the potential for the development of an adjuvant therapy for genital warts.
ABSTRACT
OBJECTIVE: Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. We compared the immune response after TCI with new construct which was derived from HPV-16 E7opt+K and pK6hf promoter instead of pCMV promoter and various adjuvant. METHODS: First, we made new construct ligated with HPV-16 E7 opt+K to Hair-follicle Specific pK6hf Promoter. Second, we applied pk6hf-E7 opt+K DNA with or without Lipofectamine 2000 and a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin of C57 BL/6 mice. To assess the ability of CTL(cytotoxic T-lymphocyte) activity, we performed intracellular cytokine staining with flow cytometric analysis to determine the number of E7-specific IFN-gamma- secreting CD8+ T cells generated in vaccinated mice with the DNA vaccine. RESULTS: Female C57BL/6 mice immunized by TCI methods with 30 microgram of pk6hf-E7 opt+K DNA with Lipofectamine2000 and CT efficiently generated E7-specific CD8(+) T cells compared with the group of pk6hf-E7 opt+K DNA only or DNA with Lipofectamine2000. CONCLUSION: Our results demonstrate that TCI of the linkaged-E7 DNA , E7 opt+K DNA to pk6hf, and Lipofectamine2000 and CT induced an antigen-specific CTL response. This result is of potential relevance for the development of therapeutic HPV-specific DNA vaccines with TCI and pK6hf promoter can be used safely.