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Objective To evaluate the value of human papillomavirus (HPV) 16/18 E6 protein detection in shunting and prognosis in patients with atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesions (LSIL). Methods A total of 98 patients with ASCUS or LSIL from the Affiliated Cancer Hospital of Shanxi Medical University between May 2014 and May 2015 were selected as the subjects. All of them received the thin-cytologic test (TCT), HPV DNA, HPV16/18 E6 protein tests and colposcopy examination. After 3-year follow-up of patients with cervical intraepithelial neoplasia (CIN) grade Ⅰor bellow lesions diagnosed by biopsy and 30 negative controls, the above tests were performed again. The efficacies of all the tests were analyzed. The value of CIN grade Ⅱ or above was predicted. Results The sensitivity, specificity, positive predictive value and negative predictive value in predicting CIN grade Ⅱor above lesions of HPV16/18 E6 protein , HPV DNA and HPV16/18 DNA was 30.8％, 95.3％, 50.0％, 90.0％, respectively; 84.6％, 37.6％, 17.2％, 94.1％, respectively and 61.5％, 67.1％, 22.2％, 91.9％, respectively in shunting study. The relative risk (RR) of CIN grade Ⅱor above lesions in patients with positive HPV16/18 E6 protein, persistent positive HPV16/18 DNA and positive HPV16/18 DNA was 13.429, 10.231 and 8.343, respectively in the follow-up study. Odds ratio (OR) of HPV16/18 E6 positive protein presenting persistent positive HPV16/18 DNA was 34.833 (95％ CI 5.020-241.711). Conclusions In patients with ASCUS and LSIL, the specificity and positive predictive value of HPV16/18 E6 protein in predicting CIN grade Ⅱ or above lesions are higher than those of HPV DNA and HPV16/18 DNA. Moreover, these patients with HPV16/18 E6 protein positive have a higher risk of developing CIN grade Ⅱ or above lesions and persistent positive HPV16/18 DNA.
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OBJECTIVE: This study was to identify small inhibitory RNAs (siRNAs) that are effective in inhibiting growth of cervical cancer cell lines harboring human papilloma virus (HPV) and to examine how siRNAs interact with interferon beta (IFN-beta) and thimerosal. METHODS: The HPV18-positive HeLa and C-4I cell lines were used. Four types of siRNAs were designed according to their target (both E6 and E7 vs. E6 only) and sizes (21- vs. 27-nucleotides); Ex-18E6/21, Ex-18E6/27, Sp-18E6/21, and Sp-18E6/27. Each siRNA-transfected cells were cultured with or without IFN-b and thimerosal and their viability was measured. RESULTS: The viabilities of HPV18-positive tumor cells were reduced by 21- and 27-nucleotide siRNAs in proportion to the siRNA concentrations. Of the two types of siRNAs, the 27-nucleotide siRNA constructs showed greater inhibitory efficacy. Sp-18E6 siRNAs, which selectively downregulates E6 protein only, were more effective than the E6- and E7-targeting Ex-18E6 siRNAs. siRNAs and IFN-beta showed the synergistic effect to inhibit HeLa cell survival and the effect was proportional to both siRNA and IFN-beta concentrations. Thimerosal in the presence of siRNA exerted a dose-dependent inhibition of C-4I cell survival. Finally, co-treatment with siRNA, IFN-beta, and thimerosal induced the most profound decrease in the viability of both cell lines. CONCLUSION: Long (27-nucleotides) siRNAs targeting E6-E7 mRNAs effectively reduce the viability of HPV18-positive cervical cancer cells and show the synergistic effect in combination with IFN-b and thimerosal. It is necessary to find the rational design of siRNAs and effective co-factors to eradicate particular cervical cancer.
Assuntos
Humanos , Linhagem Celular , Sobrevivência Celular , Células HeLa , Interferon beta , Papiloma , RNA , RNA Mensageiro , RNA Interferente Pequeno , Timerosal , Neoplasias do Colo do ÚteroRESUMO
Cervical cancer is the second most common malignant tumor of women all over the world.Infection with oncogenic human papilloma virus ( HPV) types is the most important risk factor to cause cervical cancer.The viral genome of HPV consists of three regions:the early region (E), the late region (L) and the long control region (LCR).The early proteins (E1, E2, E4, E5, E6, E7 ) play vital roles in the continuous virus expression and replication in cells.Among these proteins, E6 protein has the closest relationship with the tumorogenesis and development of cervical cancer.Many mechanisms of the HPV encoded early protein E6 in in-ducing cervical cancer have been characterized, including degradation of p53 protein, activation of telomerase, interaction with PDZ protein, disturbance of cellular signaling pathways, inhibition of immune recognition and blockage of cellular apoptosis.HPV E6 pro-tein may be a preferred target in prevention and treatment of cervical cancer.
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Objective: To study the expressions of human papillomavirus type 16/ 18-E6 (HPV16/18-E6), p53, and murine double minute-2 (MDM2) proteins in patients with laryngeal squamous cell carcinoma (LSCC) and their relationship with the development/progression of LSCC. Methods: The expressions of HPV16/18-E6, p53, and MDM2 proteins were detected by immunohistochemical method in 72 LSCC specimens, 24 specimens of precancerous larynx, 15 specimens of vocal cord polyp, and 8 normal laryngeal specimens, and their relations with clinicopathological factors of LSCC were statistically analyzed. Results: The positive rates of HPV16/18-E6 protein in the normal laryngeal, vocal cord polyp, precancerous larynx, and LSCC specimens were 0(0/8), 6.7%(1/15), 12.5%(3/24), and 37.5%(27/72); of p53 protein were 0(0/8), 13.3%(2/15), 25.0%(6/24), and 52.8%(38/72); and of MDM2 protein were 0(0/8), 6.7%(1/15), 37.5%(9/24), and 70.8%(51/72), respectively. The expressions of HPV16/18-E6, p53, and MDM2 were significantly different between the vocal cord polyp, the precacerous larynx, and the LSCC (P<0.05). The positive rates of HPV16/18-E6 and MDM2 protein in LSCC specimens increased significantly with the increase of histopathological grades (I-III) (P<0.05). The positive rates of HPV16/18-E6 protein were positively correlated with the clinical staging and lymph nodes metastasis of LSCC (P<0.05). The expression of p53 was positively correlated with the expression of HPV16/18-E6 and MDM2 in LSCC patients (P<0.05). Conclusion: Inactivationo the p53 pathway and overexpression of HPV16/ 18-E6 and MDM2 may contribute to the genesis-of human laryngeal cell carcinoma.
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Objective: To investigate the protein expression of HPV16 and its early gene E6, E7 in human laryngeal squamous cell carcinoma tissues, and to analyze their relationship with the clinical stage and pathological classification of laryngeal squamous cell carcinoma. Methods: The expression of HPV16, HPV16 E6, and HPV16 E7 protein was detected in 147 specimens of different laryngeal lesions immunohistochemically. The specimens included 82 laryngeal carcinoma, 39 non-carcinoma tissues (including 27 specimens of vocal cord polyp and 12 specimens of normal laryngeal tissues taken from more than 1.0 cm adjacent to the carcinoma), and 26 precancerous lesions (leukoplakia) of the larynx. The relationship between the protein expression with the clinical stage and histopathological classification of laryngeal squamous cell carcinoma was analyzed. Results: The positive rates of HPV16, HPV16 E6, and HPV16 E7 protein in precancerous tissues (30.77%, 26. 92%, and 26. 92%, respectively) were significantly lower than those in laryngeal carcinoma lesions (45. 12%, 39. 02%, and 42. 68%, respectively; P<0. 05 or 0. 01), but were significantly higher than those in non-carcinoma tissues (23. 08%, 5. 13%, and 2.56%, respectively; P<0.05 or 0.01). In non-carcinoma tissues, the positive rate of HPV16 protein was significantly higher than that of E6 or E7 (P<0.05 or 0.01), while there was no difference between their positive rates in laryngeal carcinoma or precancerous lesions. We found that human laryngeal carcinoma tissues with different clinical stages and different pathological classifications also had different positive rate of HPV16 protein (P<0.05), but they had a similar positive rate of HPV16 E6 and E7. Conclusion: The expression of HPV16 E6, E7 proteins after the HPV16 infection might be one of the reasons for development of human laryngeal carcinoma. Inhibition of HPV16E7 expression by immunologic strategy may have a potential for treatment of laryngeal carcinoma.