sw48 isogenic cells  (Koatech Technology Corporation)

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: HER2 expression level as a key predictive factor for the susceptibility of KRAS G13D CRC cells to CTX. A-H The inhibitory effects of CTX on cell growth ( A , C , E , and G ) and colony forming ability ( B , D , F , and H ) were evaluated in SW48 isogenic cell lines and their HER2-knockdown (KD) clones: WT_shCTRL/WT_shHER2 ( A , B ), G12D_shCTRL/G12D_shHER2 ( C , D ), G12 V_shCTRL/G12V_shHER2 ( E , F ), and G13D_shCTRL/G13D_shHER2 ( G , H ). 24 h treatment was conducted for growth inhibition ( n = 6) and a 10 day treatment for long-term anti-proliferative effects ( n = 5) at the indicated concentrations (ANOVA, ns = non-significant, **** p < 0.0001 vs. shCTRL). I The tumor growth inhibitory effect of CTX (intraperitoneal injection at 1 mg/kg every 3 days) was evaluated using xenograft mouse models of SW48 isogenic cell lines and their HER2-silenced clones ( n = 5 per group) (ANOVA, *** p < 0.001 vs. CON). J The growth inhibitory effect of CTX was tested against HCT15 (HCT15_shCTRL) and its HER2 knockdown clone (HCT15_shHER2), with caco-2 serving as a positive control ( n = 4). 24 h-treatment was conducted at the indicated concentrations. K The long-term anti-proliferative effect of CTX was assessed against HCT15_shCTRL and HCT15_shHER2 cells ( n = 5) following 10 d-treatment at the indicated concentrations (ANOVA, ns = non-significant, *** p < 0.001, **** p < 0.0001 vs. control (0), #### p < 0.0001 vs. shCTRL). L Changes in CTX sensitivity were assessed in Caco-2 and HER2-KD Caco-2 cells transduced with KRAS G13D using pcDNA4-KRAS G13D -Hismax construct following 16 h-treatment at the indicated concentrations

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Expressing, Knockdown, Clone Assay, Inhibition, Injection, Positive Control, Control, Transduction, Construct

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: Highly-expressed HER2 as a critical inducer of aggressive oncogenic features in KRAS G13D CRC cells. A-F The cell growth rate ( A , C , and E ) and colony-forming ability ( B , D , and F ) of SW48 isogenic cell lines and their HER2-knockdown (KD) clones were evaluated: G12D_shCTRL/G12D_shHER2 ( A , B ), G12V_shCTRL/G12V_shHER2 ( C , D ), and G13D_shCTRL/G13D_shHER2 ( E , F ). The cell growth rate was assessed over short-term periods at the indicated time points ( n = 5), while the colony formation rate was tested over 10 days ( n = 5) (ANOVA was used for the cell growth rate analysis and Student’s t-test for the colony formation rate, ns = non-significant, ** p < 0.01, **** p < 0.0001 vs. shCTRL). G , H The growth rates ( G ) and long-term cell proliferation rates ( H ) of HCT15 (HCT15_shCTRL) and its HER2-silenced model (HCT15_shHER2) were evaluated. For growth rate evaluation, each cell line was monitored up to 72 h ( n = 5), and cell viability at each time point was colorimetrically assessed by absorbance at 450 nm. For the cell proliferation rate assessment, both cell lines were incubated for 10 days ( n = 5) (ANOVA was used for ( G ) and Student’s t-test for ( H ), ns = non-significant, ** p < 0.01, **** p < 0.0001 vs. shCTRL). I Tumor forming abilities of SW48 isogenic cell lines and their HER2-KD clones were evaluated using xenograft mouse models. Each cell line pair (shCTRL and shHER2) was separately injected to the left or right side of the mouse flank and tumor growth was monitored for a total of 30 days ( n = 5). Both changes in tumor volume and tumor weight data were presented as relative ratios to shCTRL (Student’s t-test, ns = non-significant, ** p < 0.01, *** p < 0.001 vs. shCTRL)

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Knockdown, Clone Assay, Incubation, Injection

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: EMT as a process distinctively engaged in KRAS G13D CRCs with high HER2 levels. A GSEA was conducted on the expression dataset of KRAS G13D CRC patient samples from GSE39582 using a pre-annotated hallmark gene set collection, with FDR q < 0.05 and NOM p < 0.05 considered significant. B , C GSEA plots for the EMT gene set were generated using the expression datasets of KRAS G13D CRC ( B ) and KRAS G12 CRC ( C ) patient samples from GSE39582 . The datasets were reconstituted based on HER2 expression levels. NES, NOM p and FDR q values are as displayed above. D Representative IHC images of HER2, E-cadherin, and vimentin for HER2 high tumors with different KRAS mutational statuses are shown (I, KRAS WT ; II, KRAS G12 ; III, KRAS G12/13 ; and IV, KRAS G13D ). Images are at 100 × magnification (scale bars = 200 μm). E IHC scores of E-cadherin and vimentin were calculated by intensity score × fraction score, with quantification performed using ImageJ software. Box-and-Whisker plots were used to compare the distribution of each sample within the groups (ANOVA, ns = non-significant, ** p < 0.01). F HER2-mediated changes in the expression levels of E-cadherin and vimentin were evaluated in various SW48 isogenic cell lines. G Transwell migration assay was performed on shCTRL and shHER2 clones of SW48 isogenic cell lines, with images at 200 × magnification (scale bars = 100 μm). Quantification was conducted through ImageJ software, presenting the rates as relative ratios to shCTRL for each cell line ( n = 3) (ANOVA, ns = non-significant, ** p < 0.01, **** p < 0.0001 vs. shCTRL)

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Expressing, Generated, Software, Whisker Assay, Transwell Migration Assay, Clone Assay

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: Transcriptional regulatory axis of HER2-ELF3-KRAS as a therapeutic target for KRAS G13D CRCs. A , B The anti-proliferative effect of trastuzumab ( A ) and its impact on HER signaling ( B ) were assessed in SW48 G13D , HCT15, and LoVo cells. Trastuzumab was applied for 10 days ( A ) and 16 h ( B ) at the indicated concentrations. C , D shHER2-mediated alteration in HER signaling ( C ) and KRAS mRNA levels ( D ) ( n = 3, mean ± S.D, normalized to GAPDH) were investigated in SW48 G13D , HCT15 and LoVo cells. Cells were harvested after 36 h of incubation (ANOVA, *** p < 0.001, **** p < 0.0001, vs. shCTRL). E Schematic representation of the HER2-ELF3-KRAS transcriptional regulatory network. F Reporter gene assay was performed using pGL3-KRAS reporter gene. pGL3-KRAS was co-transfected with empty vector or pcDNA3.1-ELF3 for 24 h ( n = 4, mean ± S.D). β-Gal was used for normalization of transfection efficiency (ANOVA, ** p < 0.01, **** p < 0.0001 vs. Basic, #### p < 0.0001 vs. KRAS + emp ). G shELF3-mediated changes in the KRAS expression level were evaluated after transient transduction for 24 h. H , I shHER2-induced alterations in protein ( H ) and gene ( I ) expression level of HER2 and ELF3 were assessed ( n = 3, mean ± S.D, normalized to GAPDH ). (ANOVA, ** p < 0.01, **** p < 0.0001 vs. shCTRL). J Changes in KRAS expression levels were evaluated in SW48 G13D cells following pCDH-HER2 overexpression, shHER2 knockdown, and shELF3 transduction. HER2 overexpression and knockdown were stably induced, while shELF3 was transiently transfected for 24 h

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Incubation, Reporter Gene Assay, Transfection, Plasmid Preparation, Expressing, Transduction, Over Expression, Knockdown, Stable Transfection

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: Inhibition of ELF3-MED23 as a novel therapeutic approach to attenuate the HER2-ELF3-KRAS axis in KRAS G13D CRCs. A GST pull-down assay was performed using GST-ELF3. GST-ELF3 was co-transduced with either empty p3xFLAG or p3xFLAG-ELF3 construct. YK1 was applied 12 h post-transfection and maintained for additional 12 h prior to cell harvest. B , C YK1-mediated alterations in protein ( B ) and gene ( C ) expression levels of HER2, ELF3, and KRAS were evaluated ( n = 5, mean ± S.D, normalized to GAPDH ) . (ANOVA, **** p < 0.0001 vs. CON). D The growth inhibitory effect of YK1 was assessed in SW48 G13D and HCT15 cells. YK1 was treated for 48 h at the indicated concentrations ( n = 5) (ANOVA, **** p < 0.0001 vs. CON)

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Inhibition, Pull Down Assay, Transduction, Construct, Transfection, Expressing

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: Transcriptionally downregulating HER2 via YK1 as a relevant strategy to overcome therapeutic limitations of KRAS G13D CRC cells. A-D The effect of YK1 and CTX co-treatment on cell viability was assessed on SW48 isogenic cell lines [SW48 WT ( A ), SW48 G12D ( B ), SW48 G12V ( C ), and SW48 G13D ( D )]. Cells were treated with the indicated concentrations for 24 h (ANOVA, ns = non-significant, ** p < 0.0001, **** p < 0.0001 vs. control, ## p < 0.0001 vs. CTX). E Combination index (CI) values for YK1 (10 μM) and CTX (10 μg/mL) in SW48 isogenic cell lines were calculated using Compusyn software. F Co-treatment effects of YK1 and CTX on colony-forming ability were measured against SW48 G13D and HCT15 cells ( n = 5) following a 10 d-incubation at indicated concentrations (ANOVA, ns = non-significant, * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. control (0), ## p < 0.01, ### p < 0.01 vs. CTX). G YK1-induced changes in EMT marker gene expression were examined in SW48 G13D and HCT15 cells after 16 h of treatment at the indicated concentrations ( n = 5, mean ± S.D., normalized to GAPDH) (ANOVA, ns = non-significant, *** p < 0.001, **** p < 0.0001 vs. CON). H Transwell migration assay was performed after 16 h of YK1 treatment in SW48 G13D and HCT15 cells at the indicated concentrations. Images were captured at 200 × magnification (scale bars = 100 μm). Quantification of cell migration was conducted using ImageJ software, presented as a relative ratio on CON ( n = 3). (ANOVA, *** p < 0.001 vs. CON)

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: Control, Software, Incubation, Marker, Gene Expression, Transwell Migration Assay, Migration

Journal: Molecular Cancer

Article Title: Targeting the HER2-ELF3-KRAS axis: a novel therapeutic strategy for KRAS G13D colorectal cancer

doi: 10.1186/s12943-025-02343-5

Figure Lengend Snippet: YK1 induces potent anti-cancer effects in KRAS G13D CRC tumors. A The co-administration effect of CTX (1 mg/kg) and YK1 (10 mg/kg) was evaluated in an in vivo xenograft mouse model using SW48 G13D cells ( n = 6). Drug administration began on day 1, and tumor length and width were measured with calipers. Tumor volumes were calculated using the formula: (length × width 2 )/2. Data are presented as mean ± S.E.M. (ANOVA, ns = non-significant, **** p < 0.0001 vs. CON, #### p < 0.0001 vs. CTX). B Representative images of the excised tumors from each group ( n = 6). C Tumor weights were assessed across treatment groups ( n = 6). (ANOVA, ns = non-significant, * p < 0.05, **** p < 0.0001 vs. CON, ### p < 0.001 vs. CTX). D IHC staining for Ki67 and HER2 was performed on tumor tissues from each group. The positive area and intensity (both normalized to CON) were quantified for Ki67 and HER2, respectively ( n = 6, 10 independent fields per animal). Data are shown as mean ± S.D. (ANOVA, ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON, ### p < 0.001 vs. CTX). E , F Gene expression levels of HER2, ELF3, KRAS ( E ), and EMT markers ( F ) were evaluated in tumor tissues from the indicated treatment groups ( n = 6, mean ± S.D, normalized to GAPDH ). (ANOVA, ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON, ## p < 0.001 vs. CTX). G The combined effect of CTX (1 mg/kg) and YK1 (20 mg/kg) was tested in an in vivo orthotopic mouse model of HCT15-luc cells ( n = 3). Tumor growth was monitored for 18 days using bioluminescent imaging. Data are presented as the percentage change in tumor size relative to day 1 (mean ± S.E.M.). (ANOVA, * p < 0.05, ** p < 0.01 vs. Vehicle on day 1)

Article Snippet: Single-cell suspensions (5 × 10 6 cells) of SW48 isogenic cells were subcutaneously injected into the left flank of 5-week-old athymic nude mice (Koatech, Korea).

Techniques: In Vivo, Immunohistochemistry, Gene Expression, Imaging

Journal: PLoS ONE

Article Title: CDK1 Is a Synthetic Lethal Target for KRAS Mutant Tumours

doi: 10.1371/journal.pone.0149099

Figure Lengend Snippet: (A) Schematic of KRAS isogenic cell lines generation. KRAS mutations were introduced into the parental cell lines via r-AAV-mediated homologous recombination. A general structure of the targeting construct is represented. The resulting mutant KRAS allele is expressed from its endogenous promoter. The Neo cassette is removed from the genome of the targeted cells by Cre recombinase-mediated excision. AAV, adeno-associated virus; ITR, inverted terminal repeat; Neo, geneticin-resistance gene; P, SV40 promoter; triangles, loxP sites (Figure adapted from ). (B) RAS activation status of LIM1215 KRAS isogenic cell lines. Western blot showing active RAS (RAF1 GTP-bound) levels for LIM1215 KRAS isogenic cell lines. The RAF1 RAS binding domain (RBD) was used to precipitate GTP-RAS. The RAS activation status was tested for each clone with mutated KRAS. Precipitated RAS-GTP was detected by western blot using anti-RAS antibody. As a positive control, HeLa cells (RAS wild-type) were stimulated with epidermal growth factor (EGF) to activate the RAS pathway. HeLa and MCF7 cells (unstimulated) were used as negative controls. Total lysates were also immunoblotted with anti-β-Actin antibody as loading control. (C) and (D) KRAS dependence in the LIM1215 KRAS isogenic cell line models, obtained from the HT siRNA screen described in . Bar graph of KRAS siRNA Z-score values across the LIM1215 KRAS WT and mutant isogenic cell lines, C and D respectively. KRAS dependence was greater in the cell lines carrying KRAS mutations than in WT cells. Error bars represent SEM from three independent experiments. (E) Western blot of KRAS in SW48 cells expressing KRAS -specific siRNAs. Multiple KRAS siRNA oligos and a pool efficiently suppressed KRAS expression showing that the siRNAs were on-target.

Article Snippet: For assessment of the in vivo efficacy of AZD5438, 5x10 6 of SW620 cells, or SW48 KRAS WT or p.G12V isogenic cells were injected into the flank regions of female athymic Balb/C mice, twenty mice per cell line (Harlan Laboratories).

Techniques: Homologous Recombination, Construct, Mutagenesis, Virus, Activation Assay, Western Blot, Binding Assay, Positive Control, Control, Expressing

Journal: PLoS ONE

Article Title: CDK1 Is a Synthetic Lethal Target for KRAS Mutant Tumours

doi: 10.1371/journal.pone.0149099

Figure Lengend Snippet: (A) GTP-RAS assay showing the RAS activation status of SW48 KRAS isogenic cell lines. (B) KRAS dependence in the SW48 KRAS isogenic cell lines (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, Student’s t-test for comparison between each KRAS mutant and the WT cell lines). (C) CDK1-specific siRNAs suppress CDK1 expression. Cell viability after CDK1 depletion in SW48 isogenic KRAS cell lines (ns, not statistically significant, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, Student’s t-test for comparison between each KRAS mutant and the WT cell lines). Error bars represent SEM from three independent experiments. (D) Western blot of CDK1 in SW48 parental cells expressing CDK1-specific siRNAs.

Article Snippet: For assessment of the in vivo efficacy of AZD5438, 5x10 6 of SW620 cells, or SW48 KRAS WT or p.G12V isogenic cells were injected into the flank regions of female athymic Balb/C mice, twenty mice per cell line (Harlan Laboratories).

Techniques: Activation Assay, Comparison, Mutagenesis, Expressing, Western Blot

Journal: PLoS ONE

Article Title: CDK1 Is a Synthetic Lethal Target for KRAS Mutant Tumours

doi: 10.1371/journal.pone.0149099

Figure Lengend Snippet: (A) Exposure of SW48 isogenic cell lines to RO-3306 in a fifteen-day colony formation assay. (B) Exposure of SW48 isogenic cell lines to inhibitor AZD5438, in a fifteen-day colony formation assay. (C) Drug-dose response curves of CRC cells after AZD5438 exposure in a fifteen-day colony formation assay. ****P<0.0001, Two-way ANOVA. Error bars represent SEM of three technical replicates. All the experiments were performed two independent times with three technical replicates.

Article Snippet: For assessment of the in vivo efficacy of AZD5438, 5x10 6 of SW620 cells, or SW48 KRAS WT or p.G12V isogenic cells were injected into the flank regions of female athymic Balb/C mice, twenty mice per cell line (Harlan Laboratories).

Techniques: Colony Assay

Journal: PLoS ONE

Article Title: CDK1 Is a Synthetic Lethal Target for KRAS Mutant Tumours

doi: 10.1371/journal.pone.0149099

Figure Lengend Snippet: (A—C). CDK1 phosphorylation levels in KRAS mutant and WT cells as shown by Western blot analysis of total cell protein lysates from SW48 KRAS isogenic (A), non-isogenic pancreatic tumour cell lines (B) and non-isogenic colorectal cell lines (C). Western blots were probed for CDK1 (pThr161 CDK1 and total CDK1). β-actin detection was used as a loading control. (D and E) Bar graphs illustrating the percentage of cells in G1, S and G2/M cell cycle phases in SW48 KRAS WT or p.G12V mutant cell lines after AZD5438 exposure. SW48 KRAS WT (D) and p.G12V (E) were exposed to 0.3 μM AZD5438 or DMSO for 16, 24 and 48 hours after which cell cycle profiles were assessed by propidium iodide (PI) staining and flow cytometry. The KRAS p.G12V mutant cells showed a decrease in S and G2-fractions after exposure to AZD5438 when compared to control (DMSO) treated cells and to KRAS WT cells (AZD5438 and DMSO). (F—H) DNA synthesis in SW48 KRAS WT and p.G12V cell lines after AZD5438 exposure. (F) and (G) 5-ethynyl-2'-deoxyuridine (EDU)/ PI FACS plots in SW48 KRAS WT (F) and p.G12V mutant cells exposed to AZD5438 0.3 μM and 0.75 μM, or DMSO for 24 and 48 hours. After AZD5438 exposure, EDU/PI profiles were assessed by flow cytometry. EDU stained cells are represented in blue. (H) Bar graph illustrating the percentage of cells stained with EDU over time for both SW48 KRAS WT and p.G12V mutant cells. (I) Western blot illustrating the phosphorylation of Retinoblastoma protein (pRb) in SW48 KRAS WT and p.G12V mutant cell lines after AZD5438 exposure. Cells were exposed to AZD5438 for two hours after which total cell lysates were generated and western blotted as shown. Detection of β-Actin was used as a loading control. The levels of Rb phosphorylation on Ser807/811 were decreased in the KRAS p.G12V cells when compared to the WT cells, after AZD5438 2 hours exposure. (J) Western blot illustrating PARP1 cleavage in SW48 KRAS WT and p.G12V mutant cells after 72h of AZD5438 exposure. Cells were exposed to AZD5438 for two hours after which total cell lysates were generated and western blotted as shown. Exposure to camptothecin was used as a positive control.

Article Snippet: For assessment of the in vivo efficacy of AZD5438, 5x10 6 of SW620 cells, or SW48 KRAS WT or p.G12V isogenic cells were injected into the flank regions of female athymic Balb/C mice, twenty mice per cell line (Harlan Laboratories).

Techniques: Phospho-proteomics, Mutagenesis, Western Blot, Control, Staining, Flow Cytometry, DNA Synthesis, Generated, Positive Control