Ured by western blotting assays at 72 h posttransfection. Cologenic assays wereUred by western blotting

Ured by western blotting assays at 72 h posttransfection. Cologenic assays were
Ured by western blotting assays at 72 h posttransfection. Cologenic assays were done as described in Methods and are shown in (B). (C) Migrated and Pepstatin A clinical trials invasive cells on the lower surface of the Transwell filter were stained and photographed, 200? The number of migrated and invasive cells is shown in (D). (E) Ishikawa cells were transfected with miR-204 m, and the growth curves of Ishikawa cells were plotted by MTT assays. (F) Colony formation was observed. Transwell and cell invasion assays were done as described in Methods. The number of migrated and invasive cells is shown in (G and H). In all the panels, data are shown as mean ?SD of at least three independent experiments. * P < 0.05, ** P < 0.01; NS P > 0.05.Bao et al. Molecular Cancer 2013, 12:155 http://www.molecular-cancer.com/content/12/1/Page 11 ofFigure 6 MiR-204-5p inhibits the growth of mouse xenografts bearing human endometrial carcinoma cells. (A) IshikawaTrkB cells were transfected with lentiviruses encoding miR-204 or its scrambled control (NC). MiR-204 was detected by TaqMan PCR and normalized against U6B. Data are expressed as mean ?SD of at least three independent experiments. ** P < 0.01. (B) Tumor growth curve in nude mice bearing IshikawaTrkB cells transfected with lentiviruses encoding miR-204 or its scrambled control. Data are expressed as mean ?SD of 8 mice. * P < 0.05. (C) Photograph of tumor xenografts. (D) Tumor weights are expressed as mean ?SD of tumors from panel C. *P < 0.05. (E) Representative H E staining histopathology of miR-204 NC and miR-204 tumor tissues in mice (left panels). MiR-204 expression was detected by in situ hybridization (second panels), and TrkB, p-STAT3, Ki 67 and PCNA expression was detected by immunohistochemistry (right panels) (magnification, 200?. Results are representative of three independent experiments.protein markers ki67 and PCNA by immunohistochemical staining (Figure 6E, last two pairs of panels). The ki67 proliferation index of the NC group [(92.80 ?3.24) ] was markedly greater than that of the miR-204-5p group [(52.76 ?9.62) ; P < 0.05]. Consistently, the PCNA proliferation index of the NC group [(91.30 ?6.75) ] was significantly higher than that of the miR-204-5p group [(20.60 ?11.46) ; P < 0.05], (Additional file 4: Figure S4B). These results indicate that miR-204-5p inhibits the tumorigenicity of endometrial carcinoma cells in vivo and further suggests a tumor-suppressive effect of miR-204-5p via the TrkB/STAT3 pathway.MiR-204-5p expression correlates with tumor stage and lymph node metastasis of endometrial cancer patientsTo further determine the correlation between the clinicopathologic characteristics of endometrial cancer patients and miR-204-5p expression, we measured the expression levels of miR-204-5p in 25 normal endometrium samples and 71 endometrial cancer tissues by TaqMan PCR assays. We observed a significantly lower expression of miR-2045p in endometrial cancer tissues compared with thenormal endometrium (P < 0.0001) (Figure 7A). Moreover, Spearman correlation analysis showed a strong inverse correlation between the expression of TrkB and miR-2045p (r = -0.2414, P < 0.05) (Figure 7B). Our RT-PCR assays further showed increasingly lower miR-204-5p levels as tumors progressed from FIGO stage I to III (stage I vs. III, P < 0.05) (Figure 7C). Though miR-204-5p levels were lower in type II vs. I tumors and tumors with myometrial invasion, no statistical association was observed between miR-204-5p PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28300835 expression and.