Research Paper Volume 13, Issue 15 pp 19750—19759

Dual inhibition of FGFR4 and BCL-xL inhibits multi-resistant ovarian cancer with BCL2L1 gain

BCL2L1 gain is associated with multi-resistance. Reproduced from TCGA OvCa dataset, shown were (A) BCL2L1 expression in different copy number alteration in OvCa; Reproduced from GDSC dataset, shown were (B) volcano plot revealing drug sensitivity of cancer cells with 20q11.21 gain; (C) cancer cells with BCL2L1 gain showing resistance to 2 BCL2-targeted compounds; (D) western blotting showing BCL-xL level in 3 OvCa cell lines with different BCL2L1 status treated or untreated with BCL2L1 knockdown (KD) by shRNA; (E) cell viability detected by MTT assay in 3 OvCa cell lines over 10-day course with or without BCL2L1-KD plus cisplatin (Pt, 1 mg/mL in 0.9% NaCl); Gene enrichment analysis using (F) NET-GE platform and (G) GSEA method showing genes enriched in BCL2L1-gained OvCa cases in TCGA cohort; (H) shown was PCR validation of sgRNAs using 3 pairs of primers with Cruser method in SK-OV-3 cells prepared for CRISPR/Cas9 knockout (KO) of BCL2L1; (I) western blotting showing BCL-xL level in SK-OV-3 cells with or without BCL2L1-KO; (J) cell viability of SK-OV-3 cells with or without BCL2L1-KO plus cisplatin over 5-day course; Western blotting showing (K) levels of FGFR/NCAM family members in SK-OV-3 cells with or without BCL2L1-KO and (L) BCL2L1 (BCL-xL) level in SK-OV-3 cells with both KD of BCL2L1 and one of FGFR/NCAM family members (N = 5 in all assays; *P

Figure 2. BCL2L1 gain is associated with multi-resistance. Reproduced from TCGA OvCa dataset, shown were (A) BCL2L1 expression in different copy number alteration in OvCa; Reproduced from GDSC dataset, shown were (B) volcano plot revealing drug sensitivity of cancer cells with 20q11.21 gain; (C) cancer cells with BCL2L1 gain showing resistance to 2 BCL2-targeted compounds; (D) western blotting showing BCL-xL level in 3 OvCa cell lines with different BCL2L1 status treated or untreated with BCL2L1 knockdown (KD) by shRNA; (E) cell viability detected by MTT assay in 3 OvCa cell lines over 10-day course with or without BCL2L1-KD plus cisplatin (Pt, 1 mg/mL in 0.9% NaCl); Gene enrichment analysis using (F) NET-GE platform and (G) GSEA method showing genes enriched in BCL2L1-gained OvCa cases in TCGA cohort; (H) shown was PCR validation of sgRNAs using 3 pairs of primers with Cruser method in SK-OV-3 cells prepared for CRISPR/Cas9 knockout (KO) of BCL2L1; (I) western blotting showing BCL-xL level in SK-OV-3 cells with or without BCL2L1-KO; (J) cell viability of SK-OV-3 cells with or without BCL2L1-KO plus cisplatin over 5-day course; Western blotting showing (K) levels of FGFR/NCAM family members in SK-OV-3 cells with or without BCL2L1-KO and (L) BCL2L1 (BCL-xL) level in SK-OV-3 cells with both KD of BCL2L1 and one of FGFR/NCAM family members (N = 5 in all assays; *P < .05; **P < .01; ***P < .001; ****P < .0001).