18P - TC-1 is required for TBC1D3-induced Wnt/beta-catenin accumulation and cell migration in MCF-7 breast cancer cells

Date 17 December 2016
Event ESMO Asia 2016 Congress
Session Poster lunch
Topics Basic Science
Breast Cancer
Presenter Huzi Zhao
Citation Annals of Oncology (2016) 27 (suppl_9): ix1-ix8. 10.1093/annonc/mdw573
Authors H. Zhao
  • Immunology, Zhongda Hospital Southeast University, 210009 - Nanjing/CN

Abstract

Background

TC-1 was originally found in thyroid cancer and it was subsequently demonstrated that TC1 up-regulates Wnt/beta-catenin target genes implicated in invasiveness and aggressive behavior of cancers in many tumors. TBC1D3 is a hominoid-specific gene that was originally identified as a novel amplified oncogene, based on its ability to confer tumorigenicity to NIH 3T3 cells. TBC1D3 highly overexpresses in breast cancer, while its function in the development and progress of tumors remains unknown.

Methods

Human breast cancer MCF-7 cells were transfected with or without the Flag-TBC1D3 plasmid and then transcriptome sequencing was performed. ShRNA-NC/MCF-7 cells and shRNA-TC-1/MCF-7cells were established and identified. Then two establishment steady cells were transfected with or without the Flag-TBC1D3 plasmid, and then 10 × 104 cells of each group were performed with transwell assay treating with XAV939(Wnt/beta-catenin inhibitor) or not; other cells were lysed with RIPA and immunoblotted.

Results

The transcriptome sequencing assay showed that TBC1D3expression up-regulates TC-1 and beta-catenin mRNA levels. Western blot showed that TBC1D3 expression up-regulates TC-1 and beta-catenin protein levels. ShRNA against human TC-1knocked down TC-1 expression by more than 80% and was assessed by immunoblotting in MCF-7 cells. When TC-1 was knocked down, the TBC1D3-induced up-regulation of beta-catenin was reduced. Otherwise, there was no effect on TBC1D3-induced up-regulation of TC-1 after treating with XAV939. Transwell assay showed the migration number of MCF-7 cells transfected with TBC1D3 plasmid was 90 ± 10, which is higher than the control group, and the difference was significant (P 

Conclusions

TC-1 is required for TBC1D3-induced Wnt/beta-catenin accumulation and cell migration in MCF-7 breast cancer cells.

Clinical trial indentification

Legal entity responsible for the study

N/A

Funding

Natural Science Foundation of China

Disclosure

All authors have declared no conflicts of interest.