CRIM1 Promotes Adhesion And Migration Of Non-small Cell Lung Cancer Cell

Cysteine-rich motor neuron1 protein(CRIM1) was reported to be a type I transmenbrane protein. And it was rich in cysteine. It was reported to regulate the BMPs activity. CRIM1 was an important player in the regulation of placental development, organogenesis, angiogenesis and even kidney disease. According to these research results, herein, we proposed that CRIM1 may join in the regulation of cell-cell junction in cancer cells, and then took participate in cell adhesion and/or migration. So, in this paper we focused on its function in the cancer cell behavior. Our major work was as below:We chose different transfection conditions to find out optimal transfection condition by using q PCR and western blot to analysis the CRIM1 expression. And we found that when we hold the transfection time at 48 h and make the Vsi RNA: VRoch at 1:1, the CRIM1 si RNA could reach the highest productivity.We knockdown the CRIM1 expression, there showed a suppression of the migration ability and adhension ability respectively by migration assay and wound scratch assay in A549 cells. Samely, CRIM1 peptide treatment could up regulate the migration and adhension ability of A549 by migration assay and wound scratch assay. However, CRIM1 could not affect the A549 proliferation.Interestingly, the CRIM1 silencing A549 cells presented special morphological changes which showed similarity to the counterpart in EMT. So, we used the TGFβ1 peptide to induce A549 cells EMT, and check the CRIM1 protein expression. As we expected, it showed a significant up-regulation. The CRIM1 peptide had no capacity to induce cell EMT alone, but we found that CRIM1 could give an influence on the TGFβ1-induced-EMT, although the marker showed no significance between each other. What’s more, we found RNA interference-mediated knockdown of CRIM1 in A549 significantly increased the expression of E-CAD and N-CAD at protein level in vitro. Totally, we got the conclusion that CRIM1 regulated the E-CAD and N-CAD expression, whereas it cannot induce cell EMT.And furthermore, we overexpressed protooncogene YAP1 in A549 cells by reconstructing and transfecting the pc DNA3.1(+)-EGFP-YAP1 plasmid to found out the relationship between CRIM1 and Hippo pathway. As we expected, when overexpressed YAP1, CRIM1 showed an upregulation, which meant that CRIM1 is a target of the Hippo pathway.