Characteristic Patients (n = 36) Age (Median ± SD) 43.2 ± 15.7 Histology      Undifferentiated 21 (58.3%)    Differentiated 15 (41.7%) Primary tumor stages      T(1) 6 (16.7%)    T(2) 7 (19.5%)    T(3) 9 (25%)    T(4) 14 (38.8%) Nodular metastasis      Yes 6 (16.7%)    No 30 (83.3%) Distant metastasis      Yes 3 (8.3%)    No 33 (91.7%) PLK-1 expression      High (score 3) 17 (47.2%)    Middle (score 2) 8 (22.2%)    Low (score 1) 7 (19.5%)    Negative(score 0) 4 (11.1%) Figure buy AZD1390 1 Immunohistochemical staining of PLK-1 in human cervical carcinoma tissues. Representative

results of immunostaining are presented; cytoplasmic and some nuclear staining can be observed in tumor cells. A, Medium PLK-1 positive staining in human cervical carcinoma tissues (original magnification, 200×); B, low PLK-1 positive staining in human cervical carcinoma Cilengitide tissues (original magnification, 200×); C, PLK-1 negative control staining in human cervical carcinoma tissues (original magnification, 200×); D, Association of PLK-1 expression and primary tumor stage (* P < 0.05 compared to other group). To evaluate the possible importance of PLK-1 in tumor progression, we then evaluated the relationship between PLK-1 intensity and tumor size. Using the Spearman rank correlation test, a statistically significant positive correlation between PLK-1 expression and primary

tumor stage (r = 0.605, P = 0.002) but not metastasis was identified. Our results, therefore, provided clues that the expression of PLK-1 is Vactosertib supplier associated with the local expansion of cervical carcinoma. Levels of PLK-1

mRNA and protein in HeLa cells after PLK-1 or siRNA transfection of To evaluate the effects of PLK-1 siRNA on the biological characteristics of HeLa cells, we first transfected HeLa cells with the PLK-1 plasmid and PLK-1 siRNA. We harvested cells at different time points (0 h, 12 h, 24 h and 36 h) to measure PLK-1 gene and protein expression. As illustrated in Fig 2, levels of PLK-1 mRNA were significantly elevated after PLK-1 transfection compared to the control cells transfected with empty plasmid, with an increase in expression by 2.2-fold at 12 h, 3.5-fold at 24 h, and 4.7-fold at 36 h (P < 0.05). Similarly, an increase was also observed in protein level at 24 h (2.1-fold) and 36 h (2.3-fold). Conversely, siRNA was shown to inhibit PLK-1 mRNA and protein expression. PLK-1 mRNA levels were significantly reduced after PLK-1 siRNA transfection compared to the control cells transfected with empty plasmid, with a decrease of 49% at 12 h, 62% at 24 h, 69% at 36 h (P < 0.05). Similar decreases were also observed at the protein level at 24 h (58%) and 48 h (76%). Our results suggest that PLK-1 siRNA transfection into HeLa cells is able to knock-down the expression of PLK-1. Figure 2 Alteration of PLK-1 gene and protein expression in HeLa cells after PLK-1 or siRNA transfection. PLK-1 production in HeLa cells increased after PLK-1 transfection, but was inhibited by siRNA transfection.