Page 76 - ebook

[B. Cell Biology/Stem Cell] B-38

Functional Validation of RKIP knockout effect on Cancer Cell

Signaling

Md Abu Rayhan¹, Eun-Young Lee², Mohammad Faysal Al Mazid¹, Minjeong Kim², Hye Jin You¹,²*

¹Department of Cancer Biomedical Science, National Cancer Center (NCC)-Graduate School of Cancer Science and

Policy (GCSP), Gyeonggi 10408, Korea, ²Tumor Microenvironment Branch, Division of Cancer Biology, National

Cancer Center, Gyeonggi 10408, Korea

Colon cancer, one of common cancers is relatively well characterized. There are many pathways leading to colon
cancer malignancy including the Raf-kinase/MEK/ERK pathway. Here, we hypothesized that targeting the Raf-

kinase/MEK/ERK pathway might be targetable against tumor malignancy and selected an raf kinase regulator, RKIP
for the study. First, we generated RKIP knockout cell lines by using CRISPR/CAS9 system and validated their

expression. By gDNA PCR and immunoblotting, the expression of RKIP were confirmed in HCT116, MOCK and two
RKIP edited cell lines. In adherent culture, cell lines showed similar growth rate until confluent. Interestingly, RKIP

edited cell lines showed delayed growth until 6 days, followed robust growth in non-adherent culture, suggesting
a role of RKIP for cell plasticity. Furthermore, proteins related to EMT, such as vimentin, Snai1, and others were

altered by RKIP knockout in cell lines. we profiled transcriptome and performed gene set enrichment analysis. In
addition, we will try the detailed regulatory mechanism how RKIP is involved in cell signaling to tumor malignancy

especially modulating plasticity. In the future, we will provide biochemical evidence to clarify the role of RKIP and
its regulatory mechanism for cancer malignancy which will be helpful for anticancer therapeutic strategy of colon
cancer.