Panoply™ Human RAF1 Knockdown Stable Cell Line

Anlotinib is an effective drug for treating advanced non-small cell lung cancer (NSCLC), but resistance often develops during treatment. RAF1, a serine/threonine kinase involved in cancer progression, has limited research in NSCLC, particularly regarding anlotinib resistance. Here, bioinformatics analysis showed low RAF1 expression levels in lung cancer tissues in the TCGA and GEPIA databases. Further pathway analysis revealed a positive correlation between RAF1 expression and targeted therapy resistance, and a negative correlation with the expression of the anti-apoptotic protein Bcl-2. Immunohistochemical analysis showed that high RAF1 expression in NSCLC tissues was associated with anlotinib resistance. In vitro experiments demonstrated that RAF1 is involved in anlotinib resistance in NSCLC cells. Overexpression of RAF1 increased the viability of PC9 and PC9/AR cells and reduced apoptosis, while knockdown of RAF1 produced the opposite effect. Therefore, these results suggest that RAF1 mediates anlotinib resistance in NSCLC cells by regulating apoptosis and may serve as a predictive biomarker for anlotinib resistance in patients with advanced lung cancer.

To investigate the potential function of RAF1 in non-small cell lung cancer (NSCLC) cells, researchers constructed RAF1-overexpressing and RAF1-knockdown cells. CCK-8 assays showed that RAF1 overexpression significantly improved the survival rate of PC9 and PC9/AR cells compared to the control/pc-NC group (Figure 1A1), while RAF1 knockdown exhibited the opposite effect (Figure 1A2). Flow cytometry analysis revealed that RAF1-overexpressing cells had a decreased apoptosis rate (Figure 1B1, C1), while RAF1-knockdown cells had an increased apoptosis rate (Figure 1B2, C2). Western blot analysis showed that RAF1 overexpression reduced Bax expression in PC9 and PC9/AR cells and significantly increased Bcl-2 expression (Figure 1D1, E1). Conversely, RAF1-knockdown cells showed upregulated Bax expression and inhibited Bcl-2 expression (Figure 1D2, E2). These observations suggest that RAF1 regulates the viability and apoptosis of PC9 and PC9/AR cells.

Figure 1. Effects of RAF1 overexpression and knockdown on PC9 and PC9/AR cells viability and apoptosis. (Wu S, et al., 2025)