Abstract 24P
Background
Deregulated expression of the c-Myc oncogene is an important molecular hallmark of cancer. Previous studies have shown that the murine zinc finger protein ZF5 is a putative transcriptional regulator of c-Myc and can affect cell growth in mouse cell lines. The human zinc finger protein ZFP161 is 98% homologous to murine ZF5. The human gene Zfp161 is localized to 18p11.21 and cDNA of the gene has an open reading frame of 1347bp. We tested the regulatory effects of ZFP161 on c-Myc in human cells.
Methods
Correlation of Zfp161 and Myc in human colon cancer cells was derived from the R2 database. A Zfp161 knockout Hct116 cell line was prepared. Hct116 cells were transfected with PLK01, ZFP161 60shRNA and ZFP161 50shRNA to make Zfp161 knockdown cells. Western blot analysis was performed to determine ZFP161 and c-MYC protein levels in wild type Hct116 cells, Zfp161 knockout cells, control cells, 60sh-Zfp161 cells and in 50sh-Zfp161 cells. To test Zfp161 expression in the same cells, quantitative PCR (qPCR) analysis was performed. Clonogenic assays of 5 different cell types in vitro were performed to monitor the growth regulatory activity of Zfp161 in human cells.
Results
Western blots demonstrated decreased levels of ZFP161 protein in Zfp161 knockout and shRNA knockdown cells. Simultaneously, decreased c-MYC protein levels were observed in Zfp161 knockout and shRNA knockdown cells compared with wild type and knockdown control cells. Using qPCR, decreased c-Myc RNA levels were observed in Zfp161 knockout cells compared to wildtype cells and decreased c-Myc RNA levels were observed in shRNA knockdown cells compared to control cells. Our results show that knockout and knowdown of Zfp161 results in a significant decrease in c-Myc expression in human cells. The decrease of c-Myc in Zfp161 knockout and knockdown cells translated into a robust decrease of in vitro cell survival in colony formation assays.
Conclusions
Zfp161 is a regulator of the c-Myc oncogene in human cells. Zfp161 knockout and knowdown cells show a robust decrease of cell survival in vitro.
Clinical trial identification
Editorial acknowledgement
Legal entity responsible for the study
Zhenkun Lou.
Funding
Mayo Clinic.
Disclosure
All authors have declared no conflicts of interest.