Panoply™ Human ALOX12 Knockdown Stable Cell Line

Colorectal cancer is a highly malignant tumor with a low prognosis and mortality rate. Bevacizumab, as the first approved biologic agent for metastatic colorectal cancer (mCRC), has proven to be effective when used in combination with chemotherapy and immunotherapy. However, the efficacy of bevacizumab and immunotherapy varies greatly among colorectal cancer patients at different stages. Therefore, exploring a new biomarker to comprehensively assess the prognosis of colorectal cancer and the response to bevacizumab and immunotherapy is of great significance. Here, researchers used weighted gene co-expression network analysis (WGCNA) and receiver operating characteristic (ROC) curves to identify genes associated with bevacizumab. ALOX12 was identified as a key gene associated with bevacizumab response. Prognostic analysis and in vitro experiments further confirmed that ALOX12 is closely related to prognosis, tumor proliferation, invasion, and metastasis. Multiomics data analysis based on mutation and copy number variation (CNV) showed that RYR3 drives ALOX12 expression, while 17p12 deletion inhibits ALOX12 expression. Furthermore, the researchers explored the relationship between ALOX12 and immune cells and immune checkpoints. The results showed that high ALOX12 expression predicted higher immune infiltration and better immunotherapy response, a finding further validated using the Tumor Immune Dysfunction and Exclusion (TIDE) and Submap methods.

To verify the biological function of ALOX12 in CRC cells, researchers constructed ALOX12-knockdown HCT-116 and SW480 cell lines. CCK8 growth curves showed that downregulation of ALOX12 inhibited the proliferation of HCT-116 and SW480 CRC cells. Clonogenesis assays demonstrated that downregulation of ALOX12 significantly suppressed the colony-forming ability of HCT-116 and SW480 cells. Simultaneously, 5-ethynyl-2'-deoxyuridine (EdU) assays showed that ALOX12 knockdown reduced the proportion of positive cells. Furthermore, scratch assays showed that cell migration was inhibited in ALOX12-knockdown cells (Figure 1A-D). Transwell assays (including migration and invasion assays) also showed that the migration and invasion abilities of ALOX12-knockdown CRC cells were reduced (Figure 1E-J). Therefore, ALOX12 promotes the proliferation, invasion, and metastasis of CRC.

Figure 1. The result of wound healing (A-D) and transwell assay (E-J) in HCT-116 and SW480 CRC cell lines. (Weng S, et al., 2022)