Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: CHMP4C plasmid. Full-length cDNAs of human CHMP4C were cloned into the expression vector pCMV-MCS-3Flag.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Plasmid Preparation, Clone Assay, Expressing

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: Influence of CHMP4C expression on the occurrence and development of cervical cancer. (A) Kaplan-Meier analyses of the overall survival rate of patients with cervical squamous cell carcinoma and endocervical adenocarcinoma with high or low expression of CHMP4C, derived from the Gene Expression Profiling Interactive Analysis database. Representative immunohistochemical staining of cervical cancer tissues for CHMP4C expression: (B) Normal tissues showed weak immunostaining, (C) squamous cell carcinoma tissues showed strong immunostaining, and (D) adenocarcinoma tissue showed strong immunostaining. CHMP4C, charged multivesicular body protein 4C; TPM, transcripts per million; HR, hazard ratio.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Expressing, Derivative Assay, Gene Expression, Immunohistochemical staining, Staining, Immunostaining

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: Cell line selection and CHMP4C expression validation. (A) Expression of CHMP4C mRNA was assessed using reverse transcription-quantitative PCR in the cervical cancer SiHa, HeLa and Caski cell lines. (B) Western blotting was used to evaluate the protein expression level of CHMP4C in the cervical cancer cell lines HeLa and SiHa, and in the normal cervical H8 cell line. Experiments were repeated three times independently. *P<0.05. CHMP4C, charged multivesicular body protein 4C.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Selection, Expressing, Biomarker Discovery, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: Assessment of si-CHMP4C transfection efficiency and the effect of silencing CHMP4C expression on cervical cancer cells. After transfection with si-CHMP4C, reverse transcription-quantitative PCR assessed the expression of CHMP4C in (A) SiHa and (B) HeLa cells. The MTT assay evaluated the effect of CHMP4C silencing on the viability of (C) SiHa and (D) HeLa cells. Flow cytometry was used to assess the effect of CHMP4C silencing on the apoptosis of (E) SiHa and (F) HeLa cells. The wound-healing assay in (G) SiHa and (H) HeLa cells, and the cell invasion assay in (I) SiHa and (J) HeLa cells evaluated cell migration and invasion, respectively, in the presence of si-CHMP4C. Experiments were repeated three times independently. *P<0.05. si, small interfering; CHMP4C, charged multivesicular body protein 4C.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Transfection, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, MTT Assay, Flow Cytometry, Wound Healing Assay, Invasion Assay, Migration

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: Evaluation of CHMP4C plasmid transfection efficiency and the effect of overexpressed CHMP4C on cervical cancer cells. The overexpression efficiency of CHMP4C in (A) SiHa and (B) HeLa cells was assessed using reverse transcription-quantitative PCR. The effect of CHMP4C overexpression on the proliferation of (C) SiHa and (D) HeLa cells was evaluated using MTT assays. The effect of the overexpression of CHMP4C on the migration of (E) SiHa and (F) HeLa cells was evaluated using wound healing assays. Flow cytometry was used to assess the effect of the overexpression of CHMP4C on the apoptotic rate of (G) SiHa and (H) HeLa cells. Experiments were repeated three times independently, *P<0.05. CHMP4C, charged multivesicular body protein 4C.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Plasmid Preparation, Transfection, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Migration, Flow Cytometry

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: Effect of CHMP4C and E6 silencing on the expression of Bcl2, Bcl-XL, survivin and Caspase-7 proteins in cervical cancer cells. (A) Western blotting assessed the effect of silencing of CHMP4C expression on the expression of the apoptosis-related proteins Bcl2, Bcl-XL, Survivin and Caspase-7. Evaluation of CHMP4C knockdown using reverse transcription-quantitative PCR in (B) SiHa and (C) HeLa cells. (D) Effect of the knockdown of HPV16 E6 in SiHa cells and HPV18 E6 in HeLa cells on the expression of CHMP4C. The experiments were repeated three times independently. *P<0.05. CHMP4C, charged multivesicular body protein 4C; HPV, human papillomavirus; si, small interfering.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Expressing, Western Blot, Knockdown, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Charged multivesicular body protein 4C promotes the progression of cervical cancer through the HPV E6/miR‑543 axis

doi: 10.3892/ol.2025.15021

Figure Lengend Snippet: CHMP4C as a direct target of miR-543. Effect of the knockdown of (A) HPV16 E6 in SiHa cells and (B) HPV18 E6 in HeLa cells on the expression of miR-543. (C) Bioinformatics analysis results for a potential binding site between miR-543 and CHMP4C. (D) The dual-luciferase reporter assay was used to assess the effect of the overexpression of miR-543 on the luciferase activity of 293T cells transfected with wtCHMP4C. The overexpression efficiency of miR-543 mimics was assessed using reverse transcription-quantitative PCR in (E) SiHa and (F) HeLa cells. (G) Effect of the overexpression of miR-543 on the expression of CHMP4C was evaluating using western blotting. The experiments were repeated three times independently. *P<0.05. CHMP4C, charged multivesicular body protein 4C; miR, microRNA; mu, mutant; wt, wild-type; si, small interfering; NC, negative control; HPV, human papillomavirus.

Article Snippet: The sections were then incubated with anti-CHMP4C antibodies (1:400; #GTX122876; GeneTex, Inc.) overnight at 4°C.

Techniques: Knockdown, Expressing, Binding Assay, Luciferase, Reporter Assay, Over Expression, Activity Assay, Transfection, Reverse Transcription, Real-time Polymerase Chain Reaction, Western Blot, Mutagenesis, Negative Control