Journal: Stem cell research & therapy

Article Title: Transplantation of gastric epithelial mitochondria into human gastric cancer cells inhibits tumor growth and enhances chemosensitivity by reducing cancer stemness and modulating gastric cancer metabolism.

doi: 10.1186/s13287-025-04223-7

Figure Lengend Snippet: Fig. 1 Transplanted GES-1 gastric epithelial mitochondria suppress gastric cancer cell stemness. (A) Protein expression of stemness markers (SOX2 and NANOG) after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (B) Quantification of SOX2 and NANOG expression. (C) GRP78 expression after GES-1 mitochondrial transplantation for 24 h. (D) Quantification of GRP78 expression. (E) CD44 positive cells on MKN45 or AGS cells after GES-1 mitochon drial transplantation for 24 h. (F) Quantification of CD44 positive cells. (G) ALDH1 activity after mitochondrial transplantation for 24 h in MKN45 cells. (H) Quantification of ALDH1 activity. (I) CD24 positive cells after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (J) Quantification of CD24 posi tive cells. (K) LGR5 positive cells after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (L) Quantification of LGR5 positive cells. (M) Expression of p-JNK and JNK after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (N) NOTCH1 expression after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. The data are presented as the means ± SEMs; n ≥ 3 for independent experiments; two-tailed Student’s t test: *p < 0.05 and **p < 0.01

Article Snippet: The human g a s t r i c cancer cell line MKN45 (DSMZ No. ACC409) was purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH), under a Material Transfer Agreement ( h t t p s : / / w w w . d s m z . d e / c o l l e c t i o n / c a t a l o g u e / m i c r o o r g a n i s m s / c b d - a n d - m a t e r i a l - t r a n s f e r - a g r e e m e n t s).

Techniques: Expressing, Transplantation Assay, Activity Assay, Two Tailed Test

Journal: Stem cell research & therapy

Article Title: Transplantation of gastric epithelial mitochondria into human gastric cancer cells inhibits tumor growth and enhances chemosensitivity by reducing cancer stemness and modulating gastric cancer metabolism.

doi: 10.1186/s13287-025-04223-7

Figure Lengend Snippet: Fig. 3 Glycolytic and mitochondrial biogenesis proteins were downregulated after epithelial mitochondrial transplantation. (A) Protein expression of PKM2, MCT1, and MCT4 after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (B) Quantification of PKM2, MCT1, and MCT4 expression after GES-1 mitochondrial transplantation for 24 h. (C) Lactate metabolism were analyzed using a Lactate-Glo Assay Kit after GES-1 mitochondrial transplanta tion for 24 h in MKN45 cells. (D) Lactate secretion by MKN45 and GES-1 cells was analyzed using a Lactate-Glo Assay Kit. (E) Protein expression of PGC-1α after GES-1 mitochondrial transplantation for 24 h in MKN45 cells. (F) Quantification of PGC-1α. The data are presented as the means ± SEMs; n ≥ 3 for independent experiments; two-tailed Student’s t test: *p < 0.05 and ***p < 0.005

Article Snippet: The human g a s t r i c cancer cell line MKN45 (DSMZ No. ACC409) was purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH), under a Material Transfer Agreement ( h t t p s : / / w w w . d s m z . d e / c o l l e c t i o n / c a t a l o g u e / m i c r o o r g a n i s m s / c b d - a n d - m a t e r i a l - t r a n s f e r - a g r e e m e n t s).

Techniques: Transplantation Assay, Expressing, Glo Assay, Two Tailed Test

Journal: Stem cell research & therapy

Article Title: Transplantation of gastric epithelial mitochondria into human gastric cancer cells inhibits tumor growth and enhances chemosensitivity by reducing cancer stemness and modulating gastric cancer metabolism.

doi: 10.1186/s13287-025-04223-7

Figure Lengend Snippet: Fig. 6 Transplantation of gastric epithelial mitochondria promoted gastric cancer cell apoptosis under hypoxic conditions and enhanced chemosensitiv ity. (A) MKN45 or (B) AGS cells (C) were treated with CoCl2 and GES-1 mitochondria for 48 h. Cell viability was analyzed via an SRB assay. Protein expression and quantification of HIF-1α in (C) MKN45 or (D) AGS cells were measured after GES-1 mitochondrial transplantation under hypoxic conditions for 24 h. (E) Protein expression and quantification of Drp1 in (C) MKN45 or (D) AGS cells were analyzed after GES-1 mitochondrial transplantation under hypoxic conditions. (G) Apoptotic protein expression was quantified on the basis of the c-csapase3/caspase3 ratio via Western blotting. (H,I) Caspase activity in AGS cells was analyzed through flow cytometry. (J,K) Caspase activity in MKN45 cells was analyzed through flow cytometry. The data are presented as the means ± SEMs; n ≥ 3 for independent experiments; two-tailed Student’s t test: *p < 0.05, **p < 0.01 and ***p < 0.001; #p < 0.05, ##p < 0.01 and ###p < 0.005

Article Snippet: The human g a s t r i c cancer cell line MKN45 (DSMZ No. ACC409) was purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH), under a Material Transfer Agreement ( h t t p s : / / w w w . d s m z . d e / c o l l e c t i o n / c a t a l o g u e / m i c r o o r g a n i s m s / c b d - a n d - m a t e r i a l - t r a n s f e r - a g r e e m e n t s).

Techniques: Transplantation Assay, Sulforhodamine B Assay, Expressing, Western Blot, Activity Assay, Flow Cytometry, Two Tailed Test

Journal: Stem cell research & therapy

Article Title: Transplantation of gastric epithelial mitochondria into human gastric cancer cells inhibits tumor growth and enhances chemosensitivity by reducing cancer stemness and modulating gastric cancer metabolism.

doi: 10.1186/s13287-025-04223-7

Figure Lengend Snippet: Fig. 7 Human gastric epithelial mitochondria reduce gastric cancer cell chemoresistance by promoting apoptosis. (A,B) MKN45 or AGS cells were treated with 5-FU or GES-1 mitochondria or both in combination for 24 h. Cell viability was analyzed via SRB Assay. (C,D) Protein expression and quantification of BAX in MKN45 or AGS cells were analyzed after treatment for 72 h. (E–H) Caspase activity in MKN45 or AGS cells was analyzed through flow cytom etry. (J) Quantification of protein expression (p-AKT/AKT ratio) in MKN45 cells after GES-1 mitochondrial transplantation. The data are presented as the means ± SEMs; n ≥ 3 for independent experiments; two-tailed Student’s t test: **p < 0.01 and ***p < 0.001; #p < 0.05, ##p < 0.01 and ###p < 0.005

Article Snippet: The human g a s t r i c cancer cell line MKN45 (DSMZ No. ACC409) was purchased from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH), under a Material Transfer Agreement ( h t t p s : / / w w w . d s m z . d e / c o l l e c t i o n / c a t a l o g u e / m i c r o o r g a n i s m s / c b d - a n d - m a t e r i a l - t r a n s f e r - a g r e e m e n t s).

Techniques: Sulforhodamine B Assay, Expressing, Activity Assay, Transplantation Assay, Two Tailed Test

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Activation of EIF4E by Aurora kinase A depicts a novel druggable axis in everolimus resistant cancer cells

doi: 10.1158/1078-0432.CCR-16-2141

Figure Lengend Snippet: FLO-1 RAD-R (C1) cells (A) or SK-GT-4 RAD-R (Pool) cells (C) treated with alisertib or in combination with RAD001 were subjected to cell viability assay. Alisertib significantly inhibited the ability of RAD001 resistant cells to form colonies with or without RAD001 co-treatment. Parental FLO-1 cells (B) and SK-GT-4 cells (D) were treated with alisertib and subjected to cell viability assay. E) Human gastric MKN45 cell line displayed intrinsic resistance to RAD001 and sensitivity to alisertib as indicated by cell viability assay. Inhibition of AURKA with alisertib (F) or knockdown with siRNA (G) downregulated p-EIF4E (S209) and c-MYC protein levels as assayed by Western blotting. H) AURKA inhibition (left panel) or knockdown (right panel) significantly downregulated cap-dependent translation in MKN45 cells, as determined by a dual Renilla-firefly-luciferase pcDNA3-rLuc-PolioIRES-fluc reporter that measures cap-dependent/independent translation. I) AURKA inhibition (left panel) or knockdown (right panel) significantly downregulated c-MYC transcriptional activity in MKN45 cells, as measured by the 4xEMS luciferase reporter.

Article Snippet: MKN45 cells (5 × 10 6 ) suspended in 200 μL DMEM/growth factor-reduced Matrigel mixture (50% DMEM supplemented with 10% FBS and 50% Matrigel) were injected subcutaneously into the flank regions of female SCID Hairless Congenic (SHC) Mice (Charles River Laboratories, Inc., Stone Ridge, NY).

Techniques: Viability Assay, Inhibition, Knockdown, Western Blot, Luciferase, Activity Assay

Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

Article Title: Activation of EIF4E by Aurora kinase A depicts a novel druggable axis in everolimus resistant cancer cells

doi: 10.1158/1078-0432.CCR-16-2141

Figure Lengend Snippet: A) Animals injected with MKN45 cells, and the tumors were allowed to grow until 200 mm3 in size, then treated with RAD001, alisertib or their combination for 5 weeks. Data indicated that alisertib alone or in combination with RAD001 significantly reduced tumor size in comparison with untreated or RAD001 alone treated groups. Although tumors in RAD001 treatment alone grew at a significantly slower rate than those in untreated group, they continued to grow, confirming the intrinsic resistant phenotype to RAD001. B and C) Immunohistochemistry analysis for cleaved caspase-3 expression, marker of apoptosis, and Ki-67 expression, marker of proliferation, in representative tumors of treated groups. D) A schematic diagram showing a proposed mechanism of RAD001 resistance. AURKA expression promotes RAD001 resistance through inhibition of PP2A, which leads to activation of EIF4E. Inhibition of AURKA by alisertib restores PP2A activity, thereby inhibiting EIF4E and inducing death in RAD001 resistant cancer cells.

Article Snippet: MKN45 cells (5 × 10 6 ) suspended in 200 μL DMEM/growth factor-reduced Matrigel mixture (50% DMEM supplemented with 10% FBS and 50% Matrigel) were injected subcutaneously into the flank regions of female SCID Hairless Congenic (SHC) Mice (Charles River Laboratories, Inc., Stone Ridge, NY).

Techniques: Injection, Comparison, Immunohistochemistry, Expressing, Marker, Inhibition, Activation Assay, Activity Assay