Abstract 5987: Membrane spanning 4 domains A12 (MS4A12) c.C211T stop-gain single-nucleotide polymorphism (rs2298553) promotes metastasis in colorectal cancer via the Hippo pathway

Abstract Background: Colorectal cancer (CRC) is one of the most malignant tumors worldwide. The prognosis of patients with colorectal cancer remains poor, with a 5-year metastasis survival rate of 20%-30%. Membrane spanning 4 domains subfamily A (MS4A) is an evolving family of structurally related cell surface proteins. Previous studies demonstrated that MS4A12 expression in CRC was significantly higher than that in the normal tissues. And low expression of MS4A12 in CRC was associated with poor survival in patients. However, the impact and mechanism of MS4A12 c.C211T stop-gain single-nucleotide polymorphism (SNP) (rs2298553) on the metastatic phenotype of CRC have not previously been examined. Current opinion holds that SNP introducing premature termination codons could alter the stability and function of transcripts and proteins. The aim of this study was to investigate the correlation between MS4A12 stop-gain SNP and invasion and migration of CRC cells. Methods: Whole exome sequencing and target sequencing were used to screen SNP related to distant metastasis. Immunohistochemical staining of tissue microarray was used to detect proteins in CRC tumor cancer and normal tissues. Recombinant plasmids carrying wild-type (CC) and MS4A12 c.C211T stop-gain SNP (TT) were constructed and then transfected to colon cancer cell lines DLD1, RKO. Knockdown of MS4A12 used the short hairpin RNA (shRNA). Western blot was performed to detect the level of protein expression. Wound healing essay, migration and invasion assays were used to detect metastatic ability of CRC. RNA-seq analysis, co-immunoprecipitation and mass assays were performed to further mechanism. Results: MS4A12 c.C211T stop-gain SNP (rs2298553) was screened and identified to be associated with CRC metastasis. Tissue microarray revealed that the expression of MS4A12 was lower in the colorectal tumor tissues compared to normal tissues. The low expression of MS4A12 in CRC was associated with a poor prognosis in patients. The expression level of the TT group was close to that of the basal level of empty vector transfected groups, which suggested that the c.C211T stop-gain SNP (rs2298553) inhibited truncated MS4A12 protein expression. Compared to the CC group, the TT group showed significantly increased invasion and migration ability. Meanwhile, MS4A12 knockdown also promoted invasion and migration. GO and KEGG pathway analysis suggested that the Hippo signaling pathway might be associated with the inhibition of migration and invasion by MS4A12. Compared to the CC group, the TT group impedes YAP/TAZ phosphorylation and facilitates YAP/TAZ nuclear translocation. Conclusion: MS4A12 c.C211T stop-gain SNP (rs2298553) promotes the invasion and migration of CRC. These results provide an important theoretical basis for studying the invasion and migration of CRC. Citation Format: Shuang Liu, Ting Jiang, Yan Yuan, Haina Yu, Zhiwei Guo, Yuanhong Gao, Yan Li, Weiwei Xiao. Membrane spanning 4 domains A12 (MS4A12) c.C211T stop-gain single-nucleotide polymorphism (rs2298553) promotes metastasis in colorectal cancer via the Hippo pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5987.