Patients and samples
We collected blood samples from 78 women: patients with cervical
carcinoma (n=21; median age 44, age range 47.2 +/- 11.5 years), high
grade lesions (n= 9; median age 37, age range 42.9 +/- 16.6 years), low
grade lesions (n=24; median age 30, age range 30.2 +/- 9.6 years), and
healthy donors (n=24; median age 38.5, age range 39.25 +/- 7.2 years).
The women with premalignant lesions and cancer had their diagnosis
confirmed with colposcopy and histopathological analysis. The cancer
group included cervical adenocarcinoma and squamous cell carcinoma in
situ (stage 0), and FIGO stages I, II, III and IV. These patients were
attended at the Instituto Jalisciense de Cancerología, and the Hospital
Civil Nuevo, Guadalajara, Jalisco, México. All study participants gave
their written informed consent. This study was performed according to
the guidelines of the local institutional ethics committees and in
accordance with the 2013 Declaration of Helsinki.
Venous blood was collected from each donor (15 mL). Peripheral Blood
Mononuclear Cells (PBMCs) were separated from the blood samples using
Lymphoprep™ (STEMCELL Technologies, Vancouver, Canada) density gradient
centrifugation. The PBMCs were stored frozen in liquid nitrogen at a
temperature below –130°C until the cytometry analysis. The viability of
the thawed cells was determined with trypan blue and cells were used
when viability was greater than or equal to 90%. Serum from the blood
of the same women was isolated and stored at -80 °C.