The RT PCR and Western blotting assays showed that in miR 204i transfected cells, co transfection with siTrkB significantly reduced TrkB ex pression as compared to co transfection with siTrkB NC. Furthermore, in the absence of miR 204 5p activity, the number of colonies in Ishikawa cells was increased, while silencing of TrkB by specific siRNA resulted in a marked gefitinib mechanism of action reduction in the number of colonies compared with the scrambled control . and a similar reduction was observed in the number of migrated and invasive cells. Furthermore, we chose Ishikawa cells, who do not express TrkB to see if miR 204 5p alone exerted any functional effect on EC cell lines. As expected, cells treated with miR 204 m had no obvious change in cellular pro liferation, colony formation, migration and invasion com pared with wildtype Ishikawa cells.

Together, these results suggest that TrkB is a functionally Inhibitors,Modulators,Libraries important downstream target of miR 204 5p that is involved in the clonogenic growth, migration and invasion of endometrial carcinoma cells. MiR 204 5p inhibits the growth of mouse xenografts bearing human endometrial carcinoma cells We were interested in whether miR 204 5p suppressed the growth of xenografts bearing human endometrial carcinoma cells. We transfected IshikawaTrkB cells with miR 204 5p precursor or its scrambled control using lentiviruses. Compared to the scrambled control, mouse xenografts bearing IshikawaTrkB cells overexpressing Inhibitors,Modulators,Libraries miR 204 5p showed a dramatic reduc tion in tumor size and tumor weight.

To verify its effects on protein expression, tumor tissue was embedded in paraffin and then stained with hematoxylin and eosin for histological examination. As expected, TrkB and phospho STAT3 levels were detectable in the control xenografts. Inhibitors,Modulators,Libraries however, the miR 204 expressing tumors had significantly lower levels of these proteins, thus verifying its role as a negative regulator of the TrkB/STAT3 pathway in vivo. To determine whether miR 204 5p expression affects the in vivo proliferative ability of endometrial carcinoma cells, we examined the expression of two proliferation protein markers ki67 and PCNA by immunohistochemical staining. The ki67 proliferation index of the NC group was markedly greater than that of the miR 204 5p group. Consistently, the PCNA pro liferation index of the NC group was significantly higher than that of the miR 204 5p group.

These results indicate that miR 204 5p inhibits the tumori genicity 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 patients To further Inhibitors,Modulators,Libraries determine the correlation between the clinico pathologic characteristics of endometrial cancer patients and miR 204 5p expression, we measured Inhibitors,Modulators,Libraries the expression levels of miR 204 5p in 25 normal endometrium samples and 71 endometrial cancer tissues by TaqMan AZD-2281 PCR assays.