sn38  (MedChemExpress)

Journal: Cancer Medicine

Article Title: Maritoclax Overcomes FBW7 Deficiency‐Driven Irinotecan Resistance in Colorectal Cancer by Targeting MCL1

doi: 10.1002/cam4.71419

Figure Lengend Snippet: Loss or low expression of FBW7 promoted SN38 resistance in CRC cells. (A) The protein levels of FBW7 and MCL1 in FBW7 wild‐type (WT) and knockout (KO) HCT‐116 and DLD1 cells were detected by Western blot. CCK8 assay was performed to measure absorbance and calculate the inhibition ratio after wild‐type (WT) and FBW7 ‐ knockout (KO) (B) HCT‐116 and (C) DLD1 cells were treated by increasing doses of SN38 for 48 h. The protein levels of FBW7 and MCL1 in (D) control (Ctrl) and FBW7‐knockdown (KD) RKO and (E) scramble control (scr) and FBW7‐knockdown (shFBW7) HCT‐116 cells were detected by Western blot. CCK8 assay was performed to measure absorbance and calculate the inhibition ratio after (F) control (Ctrl) and FBW7‐knockdown (KD) RKO and (G) scramble control (scr) and FBW7‐knockdown (shFBW7) HCT‐116 cells were treated by increasing doses of SN38 for 48 h. Data are expressed as the mean ± SD ( n = 3, representing three independent experiments). p values were calculated by using Student's t ‐test; **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Irinotecan (Cat# T6228; Targetmol), SN38 (Cat# T1703; Targetmol), Marinopyrrole A (Maritoclax, Cat# T11944 ; Targetmol) and MG132 (Cat# MB5137‐1; Meilunbio) were dissolved in dimethylsulfoxide (DMSO); puromycin (Cat# MB2005; Meilunbio) was dissolved in sterile deionized water.

Techniques: Expressing, Knock-Out, Western Blot, CCK-8 Assay, Inhibition, Control, Knockdown

Journal: Cancer Medicine

Article Title: Maritoclax Overcomes FBW7 Deficiency‐Driven Irinotecan Resistance in Colorectal Cancer by Targeting MCL1

doi: 10.1002/cam4.71419

Figure Lengend Snippet: FBW7 mutation conferred resistance to SN38 in CRC cells. (A) The inhibition ratios of SN38 on FBW7 wild‐type (Lim1215, HCT‐116, Lim2405 and RKO) and mutant (LOVO, SW1463, SW48 and HCT‐8) CRC cells were detected by CCK8 assay after treatment with SN38 for 48 h. (B) The expression levels of FBW7 and MCL1 proteins in FBW7 wild‐type (Lim1215 and HCT‐116) and mutant (LOVO and SW48) CRC cells were detected by Western blot. (C) The inhibition ratio of SN38 treatment for 48 h on HA‐Vec and HA‐FBW7 R465C‐overexpressed HCT‐116 cells was detected by CCK8 assay. (D) The expression of FBW7 and MCL1 protein levels in HA‐Vec and HA‐FBW7 R465C‐overexpressed HCT‐116 cells were detected by Western blot. (E) The inhibition ratio of SN38 treatment for 48 h on FBW7 wild‐type (WT), knockout (KO), KO rescued Flag‐FBW7 wild‐type and KO rescued Flag‐FBW7 R465C HCT‐116 cells was detected by CCK8 assay. (F) The expression of FBW7 and MCL1 protein levels in cells from (E) was detected by Western blot. (G) DNA sequencing of the targeted genomic regions in CRC patient‐derived tumor tissue slides highlighting the WT and corresponding FBW7 R465C‐mutant sequences. (H) The protein levels of MCL1 in FBW7 wild‐type (WT) and R465C mutant tumor samples were detected by immunohistochemistry. (I) Three pictures of each sample were collected for quantification. Average Optical Density (%) = Integrated optical density/Area. Data are expressed as the mean ± SD ( n = 3, representing three independent experiments). p values were calculated by using Student's t ‐test or Two‐way ANOVA; ns: P > 0.05, **** p < 0.0001, ** p < 0.01, * p < 0.05.

Article Snippet: Irinotecan (Cat# T6228; Targetmol), SN38 (Cat# T1703; Targetmol), Marinopyrrole A (Maritoclax, Cat# T11944 ; Targetmol) and MG132 (Cat# MB5137‐1; Meilunbio) were dissolved in dimethylsulfoxide (DMSO); puromycin (Cat# MB2005; Meilunbio) was dissolved in sterile deionized water.

Techniques: Mutagenesis, Inhibition, CCK-8 Assay, Expressing, Western Blot, Knock-Out, DNA Sequencing, Derivative Assay, Immunohistochemistry

Journal: Cancer Medicine

Article Title: Maritoclax Overcomes FBW7 Deficiency‐Driven Irinotecan Resistance in Colorectal Cancer by Targeting MCL1

doi: 10.1002/cam4.71419

Figure Lengend Snippet: SN38 regulates MCL1 expression in CRC cells. The protein levels of MCL1 in HCT‐116 cells (A) treated with 100/200/300 nM SN38 for 48 h or (B) treated with 300 nM SN38 for 0/12/24/36/48 h were detected by Western blot. The mRNA levels of MCL1 in HCT‐116 cells (C) treated with 100/200/300 nM SN38 for 48 h or (D) treated with 300 nM SN38 for 0/12/24/36/48 h were detected by RT‐qPCR. The protein levels of MCL1 in RKO cells (E) treated with 100/200/300 nM SN38 for 48 h or (F) treated with 300 nM SN38 for 0/12/24/36/48 h were detected by Western blot. The mRNA levels of MCL1 in RKO cells (G) treated with 100/200/300 nM SN38 for 48 h or (H) treated with 300 nM SN38 for 0/12/24/36/48 h were detected by RT‐qPCR. “D” represents DMSO (vehicle control). Data are expressed as the mean ± SD ( n = 3, representing three independent experiments). p values were calculated by using Student's t ‐test, **** p < 0.0001, *** p < 0.001.

Article Snippet: Irinotecan (Cat# T6228; Targetmol), SN38 (Cat# T1703; Targetmol), Marinopyrrole A (Maritoclax, Cat# T11944 ; Targetmol) and MG132 (Cat# MB5137‐1; Meilunbio) were dissolved in dimethylsulfoxide (DMSO); puromycin (Cat# MB2005; Meilunbio) was dissolved in sterile deionized water.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: Cancer Medicine

Article Title: Maritoclax Overcomes FBW7 Deficiency‐Driven Irinotecan Resistance in Colorectal Cancer by Targeting MCL1

doi: 10.1002/cam4.71419

Figure Lengend Snippet: MCL1 overexpression resulted in SN38 resistance. Western blot was performed to detect the expression of HA‐MCL1 plasmid in (A) HCT‐116 cells and (B) RKO cells after transfection for 16 h. CCK8 assay was performed to detect the absorbance and calculate the inhibition ratio of 30/100 nM SN38 treatment for 48 h on (C) HCT‐116 cells and (D) RKO cells transfected with HA‐Vec or HA‐MCL1 plasmids. (E) The chemical structure of Marinopyrrole A (Maritoclax). Western blot was performed to detect the protein levels of MCL1 in (F) HCT‐116 cells, (G) FBW7 knockdown and control RKO cells, (H) Flag‐Vec‐ or Flag‐FBW7 R465C‐overexpressed HCT‐116 cells treated with 0.1 μM SN38 and/or 1 μM Maritoclax for 48 h. CCK8 assay was performed to detect the absorbance and calculate the inhibition ratio of 0.1 μM SN38 and/or 1 μM Maritoclax treatment for 48 h on (I) Flag‐Vec‐ or Flag‐FBW7 R465C‐overexpressed HCT‐116 cells, (J) FBW7 knockdown and control RKO cells. Data are expressed as the mean ± SD ( n = 3, representing three independent experiments). p values were calculated by using Student's t ‐test, ns: P > 0.05, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Irinotecan (Cat# T6228; Targetmol), SN38 (Cat# T1703; Targetmol), Marinopyrrole A (Maritoclax, Cat# T11944 ; Targetmol) and MG132 (Cat# MB5137‐1; Meilunbio) were dissolved in dimethylsulfoxide (DMSO); puromycin (Cat# MB2005; Meilunbio) was dissolved in sterile deionized water.

Techniques: Over Expression, Western Blot, Expressing, Plasmid Preparation, Transfection, CCK-8 Assay, Inhibition, Knockdown, Control