Imiquimod treatment effectively reduces the percentage of viable cells in a cervical carcinoma cell line but does not affect the expression of HLA-G or OCT-4.

OBJECTIVE: Cervical cancer is a challenging pathologic entity because of its lack of response to conventional chemotherapy. Imiquimod is a synthetic analogue which seems to activate skin immune cells, acting as a Toll-like receptor 7 agonist. Previous studies in the field of cervical cancer have showed that its application may play a significant role in the treatment of cervical HPV infection with or without cervical intraepithelial neoplasia (CIN). In the present study we investigate the therapeutic potential of imiquimod in a cervical carcinoma cell line and evaluate whether the expression of HLA-G and OCT-4 is altered during this treatment. METHODS: HeLa cells were cultured in Dulbecco's modified Eagle medium and treated with 200 µl of imiquimod diluted solution (50 µg/ml). Cultured cells were allocated in four groups 1) control, 2) DMSO only, 3) DMSO and imiquimod for 48 hours, 4) DMSO and imiquimod for 72 hours. RESULTS: In the imiquimod treated cell lines we observed a significant reduction of viable cells at 48 and 72 hours (p = .001). The relative expression analysis of OCT-4 and HLA-G genes at 48 and 72 hours did not reveal significant differences after imiquimod treatment. CONCLUSION: Imiquimod effectively reduces the percentage of viable HeLa cells and should be further evaluated in future clinical trials. This effect takes place as of 48 hours after its initial application and seems to persist at least until 72 hours. HLA-G and OCT-4 expression is not affected by this type of treatment.

OCT-4 and DAZL expression in precancerous lesions of the human uterine cervix.

AIM: To determine whether octamer-binding transcription factor 4 (OCT-4) and deleted in azoospermia like (DAZL) are expressed among cells with human papilloma virus (HPV) infection and cervical intraepithelial neoplasia (CIN) lesions and quantify their relative expression when compared with normal cervical cultures. METHODS: Cervical cells derived from normal cell cultures, HPV lesions and CIN lesions were cultured in Dulbecco's modified Eagle's medium supplemented with 20% amniotic fluid and 5 ng/mL basic fibroblast growth factor at 37°C and humidified 10% CO2 in air. Real-time polymerase chain reaction (PCR) was carried out using G6PD as a reference. We used REST for statistical analysis of real-time PCR. RESULTS: Whereas DAZL was not expressed either in normal cultures or HPV and CIN lesions, OCT-4 was expressed in all examined cell lines. Moreover its relative expression was significantly upregulated among cultures of HPV-infected cells (RE, 11.003; 95%CI: 0.054-36 704.527, P = 0.042), an observation that was also close to statistical significance among cultures of CIN lesions (P = 0.066). CONCLUSION: The relative expression of OCT-4 is upregulated during the early, preinvasive stages of cervical carcinogenesis. Future studies should investigate its potential as a screening marker and as a possible target of therapy.