Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: Sevoflurane induces apoptosis of glioma cells by mitochondrial apoptosis pathway. (A) JC-1 staining was performed to detect MMP level. (B) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (C) Western blotting was performed to detect the expression of Bax, cytochrome c , cleaved-caspase-9/caspase-9, cleaved-caspase-3/caspase-3 and Bcl-2 proteins. (D) Western blotting was performed to detect the expression of LC3-I, LC3-II and p62 proteins; (E) LC3-II/I ratio. **P<0.01, ***P<0.001, n=3. MMP, mitochondrial membrane potential; Bax, bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Staining, Fluorescence, Western Blot, Expressing, Membrane

Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: SEV exposure promotes apoptosis of glioma cells through upregulation of miR-211-5p expression, mediating mitochondrial apoptosis pathway. (A) U251 cells were treated with SEV and miR-211-5p mimic/mimic NC, followed by reverse transcription-quantitative PCR was performed to detect miR-211-5p in cells. (B) MTT method was performed to evaluate cell viability, while (C) flow cytometry was performed to detect cell apoptosis. (D) JC-1 staining was performed to detect MMP level. (E) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (F) Western blotting was used to detect the expression of Bcl-2, Bax, cytochrome c , cleaved-caspase-9/caspase-9 and cleaved-caspase-3/caspase-3 proteins. (G) Western blotting was used to detect the expression of LC3-I, LC3-II and p62 proteins. (H) LC3-II/I ratio. **P<0.01, ***P<0.001, n=3. SEV, sevoflurane; miR, microRNA; NC, negative control; MMP, mitochondrial membrane potential; ROS, reactive oxygen species; Bcl-2, B-cell lymphoma-2; Bax, bcl-2-associated X protein.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Flow Cytometry, Staining, Fluorescence, Western Blot, Negative Control, Membrane

Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: SIRT1 mediates sevoflurane mitochondria-dependent apoptosis pathway to induce the apoptosis of glioma cells. (A) The transfection efficiency of SIRT1 in U251 cells was elevated by reverse transcription-quantitative PCR. (B) Cell viability was assessed using the MTT method, and (C) flow cytometry detection of cell apoptosis. (D) JC-1 staining to detect MMP level. (E) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (F) Western blotting detection of Bcl-2, Bax, cytochrome c , cleaved-caspase-9/caspase-9, cleaved-caspase-3/caspase-3, and Bcl-2 protein expression. (G) Western blotting detection of LC3-I, LC3-II and p62 protein expression. *P<0.05, **P<0.01, ***P<0.001, n=3. SIRT1, silent information regulator 1; MMP, mitochondrial membrane potential; Bcl-2, B-cell lymphoma-2; Bax, bcl-2-associated X protein.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Flow Cytometry, Staining, Fluorescence, Western Blot, Expressing, Membrane

Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: Sevoflurane induces apoptosis of glioma cells by mitochondrial apoptosis pathway. (A) JC-1 staining was performed to detect MMP level. (B) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (C) Western blotting was performed to detect the expression of Bax, cytochrome c , cleaved-caspase-9/caspase-9, cleaved-caspase-3/caspase-3 and Bcl-2 proteins. (D) Western blotting was performed to detect the expression of LC3-I, LC3-II and p62 proteins; (E) LC3-II/I ratio. **P<0.01, ***P<0.001, n=3. MMP, mitochondrial membrane potential; Bax, bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Staining, Fluorescence, Western Blot, Expressing, Membrane

Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: SEV exposure promotes apoptosis of glioma cells through upregulation of miR-211-5p expression, mediating mitochondrial apoptosis pathway. (A) U251 cells were treated with SEV and miR-211-5p mimic/mimic NC, followed by reverse transcription-quantitative PCR was performed to detect miR-211-5p in cells. (B) MTT method was performed to evaluate cell viability, while (C) flow cytometry was performed to detect cell apoptosis. (D) JC-1 staining was performed to detect MMP level. (E) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (F) Western blotting was used to detect the expression of Bcl-2, Bax, cytochrome c , cleaved-caspase-9/caspase-9 and cleaved-caspase-3/caspase-3 proteins. (G) Western blotting was used to detect the expression of LC3-I, LC3-II and p62 proteins. (H) LC3-II/I ratio. **P<0.01, ***P<0.001, n=3. SEV, sevoflurane; miR, microRNA; NC, negative control; MMP, mitochondrial membrane potential; ROS, reactive oxygen species; Bcl-2, B-cell lymphoma-2; Bax, bcl-2-associated X protein.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Flow Cytometry, Staining, Fluorescence, Western Blot, Negative Control, Membrane

Journal: Molecular Medicine Reports

Article Title: Sevoflurane: A dual modulator of miR‑211‑5p and mitochondrial apoptosis in glioma therapy

doi: 10.3892/mmr.2025.13544

Figure Lengend Snippet: SIRT1 mediates sevoflurane mitochondria-dependent apoptosis pathway to induce the apoptosis of glioma cells. (A) The transfection efficiency of SIRT1 in U251 cells was elevated by reverse transcription-quantitative PCR. (B) Cell viability was assessed using the MTT method, and (C) flow cytometry detection of cell apoptosis. (D) JC-1 staining to detect MMP level. (E) MitoSOX™ Red fluorescence staining was used to detect mitochondrial reactive oxygen species levels. (F) Western blotting detection of Bcl-2, Bax, cytochrome c , cleaved-caspase-9/caspase-9, cleaved-caspase-3/caspase-3, and Bcl-2 protein expression. (G) Western blotting detection of LC3-I, LC3-II and p62 protein expression. *P<0.05, **P<0.01, ***P<0.001, n=3. SIRT1, silent information regulator 1; MMP, mitochondrial membrane potential; Bcl-2, B-cell lymphoma-2; Bax, bcl-2-associated X protein.

Article Snippet: Primary antibodies were: B-cell lymphoma-2 (Bcl-2)-associated X protein (Bax) (cat. no. 50599-2-Ig; 1:8,000; Wuhan Sanying Biotechnology); cytochrome c (cat. no. 10993-1-AP, 1:4,000; Wuhan Sanying Biotechnology); cleaved-caspase-3 (cat. no. ab32042, 1:2,000, Abcam); caspase-3 (cat. no. ab184787; 1:2,000, Abcam); caspase-9 (cat. no. ab184786; 1:2,000, Abcam); caspase-9 (cleaved Asp330) antibody (cat. no. PA5-105272; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); Bcl2 antibody (cat. no. 12789-1-AP; 1:9,000; Wuhan Sanying Biotechnology); microtubule-associated protein light chain 3 (LC3) (cat. no. 14600-1-AP; 1:2,500; Wuhan Sanying Biotechnology); P62 (cat. no. 18420-1-AP; 1:10,000; Wuhan Sanying Biotechnology); SIRT1 (cat. no. 13161-1-AP; 1:3,000; Wuhan Sanying Biotechnology); PI3K antibody (cat. no. 20584-1-AP; 1:300; Wuhan Sanying Biotechnology); phosphorylated (p-)PI3K p85/p55 (Tyr458, Tyr199) (cat. no. PA5-17387; 1:1,000; Invitrogen; Thermo Fisher Scientific, Inc.); AKT (cat. no. 10176-2-AP; 1:1,000; Wuhan Sanying Biotechnology); and p-AKT (Ser473) (cat. no. ab81283; 1:2,000; Abcam).

Techniques: Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Flow Cytometry, Staining, Fluorescence, Western Blot, Expressing, Membrane