Figure 4.

miR-142-3p negatively regulates HMGB1 levels in cervical cancer cells. A. Human cervical cancer cells were transfected with hsa-miR-142-3p mimic or negative control mimics for 48 hours. The relative expression of hsa-miR-142-3p was determined by real time PCR. ^**p<0.01, compared with mimic control transfected HeLa cells. B. HeLa cells were transfected with hsa-miR-142-3p inhibitor or negative inhibitor control for 48 hours. The relative expression of hsa-miR-142-3p was determined by real time PCR. ^**p<0.01, compared with mimic control transfected HeLa cells. (C) Human cervical cancer cell lines were transfected with miR-142-3p and negative control mimics for 24 and 48 h and HMGB1 levels were determined by western blotting. GAPDH was used as an internal reference gene. The gray value of each HMGB1 band in miR-142-3p mimic- and negative control mimic-transfected cervical cancer cells was determined using ImageJ software and was presented in a histogram. Experiments were performed in duplicate. ^**P<0.01 vs. negative control mimic-transfected cells. (D) Cervical cancer cell lines were transfected with miR-142 inhibitor and inhibitor negative controls for 24 and 48 h. HMGB1 expression levels were assessed by western blotting. The gray values in miR-142-3p inhibitor- and inhibitor negative control-transfected HeLa cells are presented in a histogram. Data are presented as the mean ± standard deviation from two experiments. ^**P<0.01 vs. negative control inhibitor-transfected HeLa cells. HMGB1, high mobility group protein B1; RT-qPCR, reverse transcription-quantitative PCR; miR-142, microRNA-142; ctrl, control.