Mouse anti-Human MTA1 monoclonal antibody, clone 5E22

To investigate the effect of metastasis-associated gene MTA1 on proliferation and invasion potential of ovarian cancer cell line A2780. The eukaryotic expressing vector pcDNA3. 1-MTA1 was introduced into A2780 cells by gene transfection in vitro. The MTA1 mRNA and protein level in cancer cells were detected by reverse transcription polymerase chain reaction (RT-PCR) and western blot, respectively. The growth activities of cancer cells were detected by trypan blue stain method. The clone formation assay in soft agar was used to observe the proliferation of cancer cells. Wound healing assay and Transwell assay were used to evaluate migration and invasion abilities of cancer cells. And the protein level of bcl-xL in ovarian cancer cells was measured by immunohistochemistry and western blot. The TUNEL assay was performed to study the apoptosis of tumor cells. Seventy-two hours after transfection, the MTA1 expression increased significantly (P < 0.01). The up-regulation of MTA1 did not affect the growth activities of cancer cells (P > 0.05), but it promoted clone formation, migration and invasion abilities of cancer cells (P < 0.01). The cellular expression of bcl-xL increased 65.22 %, with a PI value of (71.64 +/- A 5.96) %. With the up-regulation of MTA1 and bcl-xL level, the apoptotic rate of A2780 cell was decreased. MTA1 gene plays an important role in progression and metastasis of ovarian cancers, which provides an ideal strategy for gene therapy of ovarian cancers.

Objective. Though metastasis-associated protein 1 (MTAI) is widely overexpressed in human cancers and is associated with advanced clinicopathological characteristics and survival in related diseases, the association between MTAI and endometrial cancer (EC) is little known and needs to be studied. Methods. Western blot and immunohistochemistry were used to analyze protein expression level of cells and tissues, while real-time PCR was used for RNA detection. Bioinformatics tool analysis revealed the relationship between MTA1 and clinicopathological characteristics and survival. CCK-8 assay, colony-formation assay, cell scratch assay, and Transwell assay were performed to determine cell proliferation, migration and invasion abilities, respectively. Results. The expression level of MTAI was significantly higher in human EC tissues than in normal endometrium. MTAI expression was correlated positively with lymph nodes metastasis and poor survival rate in EC. Experimentally overexpressed MTA1 could promote cell proliferation, migration and invasion abilities of EC cell lines Ishikawa, HEC-1B, and RL-952, while reduction of MTAI inhibited these cell biological behaviors. Moreover, MTAI could also reverse the negative effect of miR-30c, a direct modulator of MTA1, on EC cells. Our research also revealed that overexpression of MTAI contributed to EC tumor growth, while knockdown of MTAI resulted in tumor growth inhibition. Additionally, the phosphorylation levels of mTOR (S2448) and 4E-BP1 (T37/46) changed significantly along with AKT (T308) under regulation of MTAI, both in vivo and vitro. Conclusion. Our results showed that MTAI, as a downstream target of miR-30c, might promote EC progression via AKT/mTOR/4E-BP1 pathway, which indicated the potential therapy target of MTAI in EC. (C) 2019 Published by Elsevier Inc.