Patrones de infiltración leucocitaria y de expresión tisular de quimioquinas, factor de crecimiento endotelial vascular y mediadores del metabolismo oxidativo en lesiones inflamatorias y neoplásicas de cervix

Abstract

Cervical intraepithelial neoplasia (CIN) is an epithelial lesion that could become an invasive carcinoma in a sequential manner. There are several factors that may modulate the switch from epithelial to extra epithelial neoplasia. Factors such as: increased oxidative metabolism, inflammatory events (leukocyte infiltration, chemokine production) and angiogenesis factor as vascular endothelial growth factor (VEGF) could have a role in CIN progression. Several studies have performed in order to determine the role of those factors in cervical neoplasia in relationship with the in situ or systemic productions. This arise the need to study those factor in situ and their relationship with the evolution of CIN. In the present study, the expressions of leukocyte infiltration, chemokines (MCP-1, IL-8 y MIP-1), VEGF, superoxide anion (O2-) and the production of nitric oxide (NO) were analyzed in biopsies from patients with CIN, cervix inflammation and healthy controls. Since, there is a relationship between human papilloma virus (HPV) infection and CIN; the expression of those factors in CIN will be studied according to the HPV status. Fifty five biopsies from CIN patients with different CIN grades, 7 from patients with inflammatory cervix alterations and 7 from healthy individuals were analyzed. Expressions of VEGF, chemokines and leukocyte infiltration were analyzed by indirect inmunofluorecence using appropriate monoclonal and polyclonal antibodies. The expression of O2 - was determined by histochemical procedures and NO production by ELISA. HPV infection and virus typification were determined by PCR and chemiluminescence, respectively. Increased CD3 positive cells (lymphocytes) infiltration was found in CIN related to lesion grades. Monocyte/macrophage (MФ) and neutrophil infiltrations were no significant increased. This pattern was also observed in samples from patients with inflammatory lesions. Regarding to chemokines, increased expressions of MCP-1 and IL-8 were found in CIN, no related to lesion grades. MIP-1 remained at inflammatory and control values. CD3/MCP-1 or CD3/IL-8 coexpressions were found increased in CIN, but no related to lesion grades. MФ/chemokine or neutrophil/chemokine expressions in CIN remained similar to others groups. There was no association to the grade of CIN when chemokine expressions were represented as number of positive cells, but, there was association when represented as percentage of positive cells. In this regard, CD3/MCP-1 and MФ/MCP-1 expressions decreased with the CIN evolution, however, CD3/IL-8 and MФ /IL-8 expressions increased. Expressions of MCP-1, IL-8 and MIP-1 were found increased in patients with inflammatory lesions. Epithelial and stroma VEGF expressions were increased in CIN patients related to the CIN evolution. This factor was also increased in patients with cervix inflammatory lesions. Coexpressions of CD3/VEGF and MФ/VEGF were found increased in CIN related to CIN evolution. This pattern was also found when total number of positive cells/number of patient analysis was performed. Expression of O2- was found increased in CIN related to CIN evolution; however, NO production remained similar to other groups. Expression of O2- and NO production were found no increased in biopsies from patients with inflammatory lesions. Low percentage of HPV infection (17.39%) was found in CIN patients. The higher percentage was found in patients with CIN III (60%). High risk virus infection was found in patients with CIN II and CIN III. Values from studied parameters in epithelium or stroma were not affected by HPV infection. In conclusion, the in situ expressions of studied factors and their relationship with CIN evolution, suggest that lymphocyte infiltration, increased expressions of chemokines, VEGF and oxidative stress could be important factors in the evolution of CIN with further cervical carcinoma. Further studies are needed to determine the evolution of those factors from CIN to cervical carcinoma.