Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: Knockdown of SKA2 inhibits gastric cancer growth in vitro and in vivo (A) SKA2 mRNA expression levels in gastric cancer from the TCGA database via the GEPIA2 portal. ∗ p < 0.05. (B) The KM-Plotter database was used to demonstrate the impact of high SKA2 expression on the overall survival (OS) of patients with gastric cancer. (C–E) Relative cell growth of SNU638, NUGC3, and SNU668 SKA2-knockdown cell lines was detected using CCK-8 assay ( n = 5). (F–H) Colony formation assay showed inhibited viability after SKA2-knockdown in SNU638, NUGC3, and SNU668 cell lines compared to control. Representative images are shown from 3 biologically independent experiments. (I–K) Relative cell growth of SKA2 overexpression in SNU638, NUGC3, and SNU668 SKA2-knockdown cell lines was detected using CCK-8 assay ( n = 5). (L) NUGC3 scramble cells and NUGC3 SKA2-knockdown cells were injected subcutaneously into BALB/c-nude mice ( n = 5 mice per group). Three weeks after injection, xenografts were removed. Representative images of xenografts were shown. (M) Tumor volume and (N) tumor weight were determined. Data in (C)–(E), (I)–(K), (M), and (N) are presented as mean ± SD, n = 5 biologically independent samples. Data in (F)–(H) are presented as mean ± SD, n = 3 biologically independent experiments. p values in (C)–(E), (I)–(K), and (M) are based on two-factor repeated measures ANOVA. p values in (A), (F)–(H), and (N) are based on Student’s t test or one-way ANOVA (∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001).

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: Knockdown, In Vitro, In Vivo, Expressing, CCK-8 Assay, Colony Assay, Control, Over Expression, Injection

Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: Knocking down SKA2 induces gastric cancer cell lines G2/M arrest (A and B) Cell cycle analysis of SKA2 knockdown in SNU638 and NUGC3 cell lines. Percentage of flow cytometric histogram images shows the effects of SKA2. (C) Western blotting analysis of the expression of Cyclin D1, Cyclin A2, Cyclin B1, and SKA2 in SNU638 and NUGC3 SKA2-knockdown cell lines. α-Tubulin was used as the internal control. (D and E) Cell cycle analysis of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines. Percentage of flow cytometric histogram images shows the effects of SKA2. (F) Western blotting analysis of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines using anti-Cyclin D1, anti-Cyclin A2, anti-Cyclin B1, and anti-SKA2 antibodies. α-Tubulin was used as the internal control. Representative flow cytometry histograms and blotting images are shown from 3 biologically independent experiments. Data in (A)–(B) and (D)–(E) are presented as mean ± SD, n = 3 biologically independent experiments. p values are based on a one-way ANOVA test. ns, no significance (∗∗ p < 0.01; ∗∗∗ p < 0.001).

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: Cell Cycle Assay, Knockdown, Western Blot, Expressing, Control, Over Expression, Flow Cytometry

Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: Knocking down SKA2 induces gastric cancer cell line apoptosis (A and B) Effects of SKA2 knockdown on apoptosis in SNU638 and NUGC3 cell lines. Representative flow cytometric plots are shown. (C) Western blotting analysis of the expression of PARP, Cleaved-Caspase3, and SKA2 in SNU638 and NUGC3 SKA2-knockdown cell lines. α-Tubulin was used as the internal control. (D and E) Effects of SKA2 overexpression on apoptosis in SNU638 and NUGC3 SKA2-knockdown cell lines. Representative flow cytometric plots are shown. (F) Western blotting analysis of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines using anti-PARP, anti-Cleaved-Caspase3, and anti-SKA2 antibodies. α-Tubulin was used as the internal control. Representative flow cytometry plots and blotting images are shown from 3 biologically independent experiments. Data in (A)–(B) and (D)–(E) are presented as mean ± SD, n = 3 biologically independent experiments. p values are based on a one-way ANOVA test (∗∗∗ p < 0.001).

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: Knockdown, Western Blot, Expressing, Control, Over Expression, Flow Cytometry

Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: Identification and enrichment analyses of DEGs associated with SKA2 (A) The number of up-regulated and down-regulated DEGs. (B) Heatmap plot for hierarchical cluster analysis of all sequencing samples and DEGs. (C) Volcano plot for the distribution of DEGs. (D) GO enrichment analysis results for DEGs. (E) Heatmap plot of top 20 genes ranked in metabolic process. (F and G) Real-time PCR detection of SKA2 and SLC6A9 in SNU638 and NUGC3 SKA2-knockdown cell lines. (H) Western blotting analysis of the expression of SLC6A9/Glyt1 and SKA2 in SNU638 and NUGC3 SKA2-knockdown cell lines. α-Tubulin was used as the internal control. (I and J) Real-time PCR detection of SKA2 and SLC6A9 in SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines. (K) Western blotting analysis of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines using anti-SLC6A9/Glyt1 and anti-SKA2 antibodies. α-Tubulin was used as the internal control. Representative blotting images are shown from 3 biologically independent experiments. Data in (F)–(G) and (I)–(J) are presented as mean ± SD, n = 3 biologically independent experiments. p values are based on a one-way ANOVA test (∗∗∗ p < 0.001).

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: Sequencing, Real-time Polymerase Chain Reaction, Knockdown, Western Blot, Expressing, Control, Over Expression

Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: The intracellular GSH and ROS level affected by SKA2 and SLC6A9/Glyt1 in GC (A–E) Intracellular glycine, serine, threonine, methionine, and glutathione levels in SKA2-shRNA/SNU638 and scramble-shRNA/SNU638 cells. (F and G) Relative DCFH-DA fluorescence measured by flow cytometry of cells treated with SKA2 shRNA or scrambled shRNA. (H and I) Relative DCFH-DA fluorescence measured by flow cytometry of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines. (J) Western blotting analysis of GLYT1 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines using anti-Flag and anti-SKA2 antibodies. α-Tubulin was used as the internal control. (K) Relative DCFH-DA fluorescence measured by flow cytometry of Glyt1 overexpression in SNU638 SKA2-knockdown cell lines. (L) Relative DCFH-DA fluorescence measured by flow cytometry of Glyt1 overexpression in NUGC3 SKA2-knockdown cell lines. (M) Cell cycle analysis of GLYT1 overexpression in SNU638 SKA2-knockdown cell lines. (N) Effects of GLYT1 overexpression on apoptosis in SNU638 SKA2-knockdown cell lines. Representative flow cytometry histograms and blotting images are shown from 3 biologically independent experiments. Data are presented as mean ± SD, n = 3 biologically independent experiments. p values in (A)–(E) are based on Student’s t test. p values in (F)–(N) are based on a one-way ANOVA test (ns, no significance; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001).

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: shRNA, Fluorescence, Flow Cytometry, Over Expression, Knockdown, Western Blot, Control, Cell Cycle Assay

Journal: iScience

Article Title: SKA2 promotes gastric cancer progression by regulating glutathione metabolism

doi: 10.1016/j.isci.2026.115202

Figure Lengend Snippet: Knocking down SKA2-induced cell-cycle arrest and apoptosis through the SKA2/ROS/ATM axis in GC cell lines (A) Western blotting analysis of γ-H2AX (Ser139), ATM, p -ATM (Ser1981), p -Chk2 (Thr68), and SKA2 expression in SNU638 and NUGC3 SKA2-knockdown cell lines. (B) Western blotting analysis of SKA2 overexpression in SNU638 and NUGC3 SKA2-knockdown cell lines using antibodies against γ-H2AX (Ser139), ATM, p -ATM (Ser1981), p -Chk2 (Thr68), and SKA2. (C) Western blotting analysis of KU-55933 treatment in SNU638 and NUGC3 SKA2-knockdown cell lines using antibodies against cyclin D1, cyclin A2, cyclin B1, p -Chk2 (Thr68), PARP, Cleaved-Caspase3, JNK, p -JNK (Thr183/Tyr185), ATM, p -ATM (Ser1981), and SKA2. (D) Western blotting analysis of BML-277 treatment in SNU638 and NUGC3 SKA2-knockdown cell lines using antibodies against cyclin D1, cyclin A2, cyclin B1, p -Chk2 (Thr68), and SKA2. (E) Western blotting analysis of P38, p-P38 (Thr180/Tyr182), ERK, p -ERK1/2 (Thr202/Tyr204), JNK, p -JNK (Thr183/Tyr185), and SKA2 expression in SNU638 and NUGC3 SKA2-knockdown cell lines. (F) Western blotting analysis of the rescue effect of SKA2 overexpression on MAPK pathway markers (ERK, p -ERK1/2, JNK, and p -JNK) in SNU638 and NUGC3 SKA2-knockdown cell lines. (G) Western blotting analysis of JNK-IN-8 treatment in SNU638 SKA2-knockdown cell lines using antibodies against PARP, cleaved-caspase3, JNK, p -JNK (Thr183/Tyr185), and SKA2. α-Tubulin was used as the internal control for all blots. Representative blotting images are shown from 3 independent experiments.

Article Snippet: Human gastric cancer cell lines SNU638 and SNU668 were obtained from the Korean Cell Line Bank (KCLB, Seoul, Korea)., NUGC3 cells were obtained from the Japanese Collection of Research Biosources (JCRB, Osaka, Japan).

Techniques: Western Blot, Expressing, Knockdown, Over Expression, Control