Construction and identification of human ETV4 deletion mutants

Abstract:

Objective：To construct ETS Variant 4（ETV4） truncated mutants and to investigate their role in colorectal cancer cell growth and migration. Methods：Based on our previous constructed ETV4 vector，ETV4 deletion mutants that containing different con－served domains were constructed：ETV4（1-339） and ETV4（340-484）. Each mutant function was analyzed in colorectal cancer cell growth by CCK-8 kit. Each mutant function in colorectal cancer cell migration was detected with wound healing assay. Moreover，each mutant function in EMT was further measured by qRT-PCR. Results：CCK-8 assay showed that mutant ETV4（1-339） didn’t promote cancer cell growth whereas mutant ETV4（340-484） and wide type ETV4 evidently promoted cancer cell growth. Wound healing assay revealed that mutant ETV4（340-484） and wide type ETV4 promoted colorectal cancer cell migration. Further qRT-PCR analysis showed that mutant ETV4（340-484） and wide type ETV4 were able to alter the expression of ETM markers whereas mutant ETV4（1-339） could not. Conclusion：ETS domain of ETV4 is indispensable in ETV4-induced colorectal cancer cell prolif－eration and migration，which is probably related with EMT.