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Objective To evaluate the diagnostic accuracy and efficacy on cervical precancerous lesions and cervical cancer by HPV 16/18 E6 protein detection. Methods A total of 439 females with sexual activities were selected from Department of Gynaecology in Shanxi Cancer Hospital from May 2014 to January 2015, including 299 cases of cervical intraepithelial neoplasia (CIN)Ⅱ, CINⅢ or cervical cancer (the case group), and the other 140 cases (the control group). All the patients accepted the thinprep cytology test (TCT), HPV DNA and HPV 16/18 E6 oncoprotein tests and colposcope examination. Results The positive rates of the TCT, HPV DNA, HPV 16/18 E6 oncoprotein in the case group were 97.0 % (290/299), 94.3 % (282/299) and 66.9 % (200/299), respectively, and those in the control group were 44.3 % (62/140), 21.4 % (30/140) and 2.9%(4/140), respectively, and there were significant differences between both groups (all P<0.05). The sensitivity and specificity of the HPV 16/18 E6 oncoprotein test in detecting CINⅡ and above were 66.9 %and 97.1 %, respectively, and both of HPV DNA test were 94.3 % and 78.6 %, respectively; The consistent rate between HPV 16/18 E6 and HPV DNA was 71.9 % (κ= 0.21). In the case group, when TCT was associated with HPV DNA test, the sensitivity, specificity and accuracy were 98.9%, 82.8%and 81.7%, respectively, and when TCT was combined with HPV 16/18 E6 oncoprotein test, those were up to 97.9 %, 97.1 % and 95.0 %. Conclusion HPV 16/18 E6 oncoprotein test can improve the specificity of cervical cancer screening, so it may be used as a primary screening method in the less developed areas where HPV DNA test is difficult to be carried out, or as a shunt method for HPV DNA positive patients, which will allocate the limited health resources rationally.
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Objective To explore the relationship of signal transduction among human papillomavirus 18 E6 oncoprotein (HPV18E6), signal transducers and activators of transcription 1 (STAT1), protein kinase R( PKR )/α subunit of eukaryotic initiation factor 2 ( eIF2α ), nuclear factor-kappa Bp65 ( NF-κBp65 ), mitogen-activated protein kinase( MAPK)/c-Jun N-terminal kinase(JNK) ,and possible molecular mechanism. Methods Construct two lentiviral vectors which contain shRNA interfering sequence aiming at the targets of HPV18E6 oncogene and NC sequence( HPV18E6-RNAi-LV, NC-GFP-LV), based on the transduction with HPV18E6-RNAi-LV and NC-GFP-LV into HeLa cell to interfere the expression of HPV18E6 oncogene and NC sequence,the expressions of mRNA and protein( including phosphating patem)of HPV18E6, STATI, PKR, eIF2α, NF-κBp65, MAPK, JNK are measured with RT-PCR and Western blot, the difference of proliferation and sensitivity to carboplatin of HeLa cell are determined with Transwell cell methods and MTT among every groups. Results The expression of HPV18E6 oncogene can affect the expression level of mRNA and protein of NF-κBp65 and PKR genes, also affect phosphating levels of phosphating protein p-STAT1, p-PKR and p-eIF2α;the restraining rates of proliferation and sensitivity to carboplatin of HeLa cell are higher in HPV18E6-RNAi-LV group than the other groups( P＜0. 05 or P＜0.01 ). Conclusion HPV18E6 oncoprotein not only reduces the expression of PKR but dephosphorylates p-STAT1, pPKR and p-eIF2α to restrain activation of PKR/eIF2α signal transduction passage, maintain the proliferation and invading ability of HeLa cell and restrain apoptosis. The signal transduction among HPV18E6, MAPK/JNK are not clear.
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Objective: To evaluate the role of HPV16E6,cyclin D1 ,and human telomerase transcriptase (hTERT) in the development and progression of nasopharyngeal carcinoma ( NPC ) and to discuss the clinical significance. Methods: Immunohistochemistry was used to detect the expression of HPV16E6, cyclin D1, and hTERT in paraffin-embedded nasopharyngeal carcinoma tissues and nasopharyngeal chronic inflammation tissues. The relationship between their expression with the clinicopathological features of NPC was analyzed; the influence of their expression on prognoses of patients was also analyzed. Results: The positive rates of HPV16E6,cyclin D1 ,and hTERT in NPC tissues were 62. 5%(35/56) ,50. 0%(28/56) , and 67. 9%(38/56) ,respectively, which were significantly higher than those in the inflammation tissues (P0. 05). HPV16E6 expression was positively correlated with cyclin D1(r=0. 480,P<0. 001) and hTERT (r=0. 494,P<0. 001)in NPC tissues. The mean survival period and median survival period in HPV16E6,cyclin D1 and hTERT positive patients were lower than those in the negative ones (P<0. 05). Conclusion: High expression of HPV16E6,cyclin D1 ,and hTERT might be involved in the development and progression of NPC. HPV16E6 may interact with cyclin D1 and hTERT, contributing to the development of NPC. Examination of the three agents may help to predict the prognoses of NPC.
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OBJECTIVE: Adeno-associated virus Rep 78 protein is known to inhibit the promoter site of several onco-genes and viral gene, including the human papillomavirus type 16 E6 transforming genes. In this study, we investigated AAV Rep 78 mediated inhibition of HPV 16 E6 promotor activity. METHODS: pcDNA3.1/V5/His-Topo vector, cloned by AAV mediated Rep 78, is transfected into cervical cancer cell line (Caski). After that, we confirmed HPV16 derived E6 expression and cell growth inhibition. RESULTS: Transfection rate of Rep78 GFP-vector, approximately from 30 to 60 per-cent, is highly expressed at first day. But E6 expression is lower at this day. The growth of CaSki and HeLa cervical cancer cell lines was inhibited by Rep78 (p<0.05). But, the other cervical cancer cells were unaffected by Rep78 transfection. CONCLUSION: In spite of the high Rep78 transfection efficiency and expression rate, we could not show the cervical cancer cell growth inhibition. In our data, long term expression of Rep78 strategy is needed for cervical carcinoma gene therapy using adeno-associated virus vector.
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Humans , Cell Line , Clone Cells , Dependovirus , Genes, Viral , Genetic Therapy , Human papillomavirus 16 , Oncogenes , Transfection , Uterine Cervical NeoplasmsABSTRACT
Objective:To evaluate the role of HPV16E6,cyclin D1,and human telomerase transcriptase(hTERT) in the development and progression of nasopharyngeal carcinoma(NPC) and to discuss the clinical significance.Methods: Immunohistochemistry was used to detect the expression of HPV16E6,cyclin D1,and hTERT in paraffin-embedded nasopharyngeal carcinoma tissues and nasopharyngeal chronic inflammation tissues.The relationship between their expression with the clinicopathological features of NPC was analyzed;the influence of their expression on prognoses of patients was also analyzed.Results: The positive rates of HPV16E6,cyclin D1,and hTERT in NPC tissues were 62.5%(35/56),50.0%(28/56),and 67.9%(38/56),respectively,which were significantly higher than those in the inflammation tissues(P0.05).HPV16E6 expression was positively correlated with cyclin D1(r=0.480,P
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OBJECTIVE: To evaluate the correlation of HPV infection and E6 oncoprotein expression with carcinogenesis and prognostic role of cervical carcinoma METHODS: Paraffin embedded tissues were obtained from 35 invasive cervical carcinoma, 15 carcinoma in situ, 12 dysplasia of cervix and 20 patients with benign uterine disease as control. HPV type was determined by polymerase chain reaction using type specific primers. E6 oncoprotein expression was analyzed by immunohistochemical staining using C1P5 mouse monoclonal antibody. RESULTS: 1. The positivity of HPV type 16 PCR in invasive cervical carcinoma was 77.1% and it was significantly higher than carcinoma in situ(53.3%), cervical dysplasia(25%), and control (10%). But the positivity of HPV type 18 was not correlated between above groups. 2. The positivity of HPV type 16/18 in cervical carcinoma was not correlated to patient's age. 3. The positivity of HPV in cervical carcinoma was correlated to clinical stage of cervical carcinoma but the positivity of HPV in cervical dysplasia was not correlated to the degree of dysplasia. 4. The positivity of E6 oncoprotein expression was 77.1% in invasive cervical carcinoma, 66.7% in carcinoma in situ and 20% in control group. E6 oncoprotein was not decteted among the cases which negative was for HPV 16/18. The E6 oncoprotein expression in 35 cases of invasive cervical carcinoma was negative in 8, weakly positive in 7, positive in 12, and strong positive in 8 ases. But in 20 cases of normal control group, there was no case of strong positive or posirive E6 oncoprotein expression. 5. The E6 oncoprotein expression was significantly correlated to the clinical stage and lymph node metastasis. The incidence of lymph node metastasis increased and the stage became higher as the E6 oncoprotein expression increased. CONCLUSION: These results suggest that HPV is one of the most important factors in the carcinogenesis of cervical carcinoma and the E6 oncoprotein expression may be associated with biological aggressiveness of cervical carcinoma.