Abstract Aberrant cholesterol biosynthesis is a notable hallmark of cancers, supporting unlimited cell proliferation. Previously, we delineated that cholesterol synthases FDFT1 and SQLE promoted colorectal cancer (CRC) cell proliferation, while how these enzymes influence CRC metastasis and stemness remains unclear. This study demonstrates that suppress of FDFT1 or SQLE enhances CRC metastasis and stemness through impairing ER stress and degrading PDIA4 or SYVN1 individually.