IKKε phosphorylates kindlin-2 to induce invadopodia formation and promote colorectal cancer metastasis.

Abstract

Invadopodia formation is a key driver of cancer metastasis. The noncanonical IkB-related kinase IKKε has been implicated in cancer metastasis, but its roles in invadopodia formation and colorectal cancer (CRC) metastasis are unclear. Methods: Immunofluorescence, gelatin-degradation assay, wound healing assay and transwell invasion assay were used to determine the influence of IKKε over-expression, knockdown and pharmacological inhibition on invadopodia formation and the migratory and invasive capacity of CRC cells in vitro. Effects of IKKε knockdown or pharmacological inhibition on CRC metastasis were examined in mice. Immunohistochemistry staining was used to detect expression levels of IKKε in CRC patient tissues, and its association with prognosis in CRC patients was also analyzed. Immunoprecipitation, western blotting and in vitro kinase assay were constructed to investigate the molecular mechanisms. Results: IKKε co-localizes with F-actin and the invadopodia marker Tks5 at the gelatin-degrading sites of CRC cells. Genetic over-expression/knockdown or pharmacological inhibition of IKKε altered invadopodia formation and the migratory and invasive capacity of CRC cells in vitro. In vivo, knockdown or pharmacological inhibition of IKKε significantly suppressed metastasis of CRC cells in mice. IKKε knockdown also inhibited invadopodia formation in vivo. Clinical investigation of tumor specimens from 191 patients with CRC revealed that high IKKε expression correlates with metastasis and poor prognosis of CRC. Mechanistically, IKKε directly binds to and phosphorylates kindlin-2 at serine 159; this effect mediates the IKKε-induced invadopodia formation and promotion of CRC metastasis. Conclusions: We identify IKKε as a novel regulator of invadopodia formation and a unique mechanism by which IKKε promotes the metastasis of CRC. Our study suggests that IKKε is a potential target to suppress CRC metastasis.

• Authors: Ge Liu, Yantao Bao, Chaohua Liu, Qinchang Zhu, Lin Zhao, Xiaopeng Lu, Qian Zhu, Yafei Lv, Feng Bai, He Wen, Yujie Sun, Wei-Guo Zhu
• Journal: Theranostics (Theranostics) Vol. 10 Issue 5 Pg. 2358-2373 ( 2020) ISSN: 1838-7640 [Electronic] Australia
• PMID: 32104508 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © The author(s).

Chemical References

• Actins
• Cytoskeletal Proteins
• Fish protein, mouse
• Muscle Proteins
• Phosphate-Binding Proteins
• RNA, Small Interfering
• kindlin-2 protein, mouse
• I-kappa B Kinase

Topics

• Actins (metabolism)
• Animals
• Cell Line, Tumor
• Cell Movement
• Cell Proliferation
• Colorectal Neoplasms (genetics, metabolism, pathology)
• Cytoskeletal Proteins (genetics, metabolism)
• Female
• Gene Knockdown Techniques
• Humans
• I-kappa B Kinase (genetics, metabolism)
• Male
• Mice
• Muscle Proteins (genetics, metabolism)
• Neoplasm Metastasis
• Phosphate-Binding Proteins (metabolism)
• Phosphorylation
• Podosomes (genetics, metabolism)
• RNA, Small Interfering (genetics)