human cervical cancer epithelial cells  (ATCC)

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 HPV16-positive cervical cancer SiHa cell line (National Infrastructure of Cell Line Resource) was cultured in RPMI1640 medium (HyClone; Cytiva), and the human cervical epithelial immortalized H8 (Jennio Biotech Co., Ltd.), HPV16-positive cervical cancer Caski (National Infrastructure of Cell Line Resource) and HPV18-positive cervical cancer HeLa (Pricella ® ; Wuhan Elabscience Biotechnology Co., Ltd.) cell lines were cultured in DMEM medium (HyClone; Cytiva), containing 10% fetal bovine serum (Shanghai Excell Biological Technology Co., Ltd.).

Techniques: Selection, Expressing, 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 HPV16-positive cervical cancer SiHa cell line (National Infrastructure of Cell Line Resource) was cultured in RPMI1640 medium (HyClone; Cytiva), and the human cervical epithelial immortalized H8 (Jennio Biotech Co., Ltd.), HPV16-positive cervical cancer Caski (National Infrastructure of Cell Line Resource) and HPV18-positive cervical cancer HeLa (Pricella ® ; Wuhan Elabscience Biotechnology Co., Ltd.) cell lines were cultured in DMEM medium (HyClone; Cytiva), containing 10% fetal bovine serum (Shanghai Excell Biological Technology Co., Ltd.).

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 HPV16-positive cervical cancer SiHa cell line (National Infrastructure of Cell Line Resource) was cultured in RPMI1640 medium (HyClone; Cytiva), and the human cervical epithelial immortalized H8 (Jennio Biotech Co., Ltd.), HPV16-positive cervical cancer Caski (National Infrastructure of Cell Line Resource) and HPV18-positive cervical cancer HeLa (Pricella ® ; Wuhan Elabscience Biotechnology Co., Ltd.) cell lines were cultured in DMEM medium (HyClone; Cytiva), containing 10% fetal bovine serum (Shanghai Excell Biological Technology Co., Ltd.).

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 HPV16-positive cervical cancer SiHa cell line (National Infrastructure of Cell Line Resource) was cultured in RPMI1640 medium (HyClone; Cytiva), and the human cervical epithelial immortalized H8 (Jennio Biotech Co., Ltd.), HPV16-positive cervical cancer Caski (National Infrastructure of Cell Line Resource) and HPV18-positive cervical cancer HeLa (Pricella ® ; Wuhan Elabscience Biotechnology Co., Ltd.) cell lines were cultured in DMEM medium (HyClone; Cytiva), containing 10% fetal bovine serum (Shanghai Excell Biological Technology Co., Ltd.).

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 HPV16-positive cervical cancer SiHa cell line (National Infrastructure of Cell Line Resource) was cultured in RPMI1640 medium (HyClone; Cytiva), and the human cervical epithelial immortalized H8 (Jennio Biotech Co., Ltd.), HPV16-positive cervical cancer Caski (National Infrastructure of Cell Line Resource) and HPV18-positive cervical cancer HeLa (Pricella ® ; Wuhan Elabscience Biotechnology Co., Ltd.) cell lines were cultured in DMEM medium (HyClone; Cytiva), containing 10% fetal bovine serum (Shanghai Excell Biological Technology Co., Ltd.).

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

Journal: bioRxiv

Article Title: VP-CLEM-Kit: An accessible pipeline for visual proteomics using super resolution volume correlative light and electron microscopy (SR-vCLEM)

doi: 10.1101/2025.04.08.647770

Figure Lengend Snippet: (a) Overview RF and WF images showing serial sections of HeLa cells (scale bar: 400 µm) with insets showing zooms of one of the sections in each modality (scale bar: 200 µm). (b, c, d) High resolution LSM900 Airyscan images showing the preservation of Hoechst (nucleus), mitoGFP (mitochondria) and lysotracker (lysosomes) in a 200 nm thick easyIRF section (scale bar: 20 µm). (e) Cell of interest showing overlay of Hoechst, mitoGFP and lysotracker signals and (f) ultrastructure (scale bar: 10 µm), with inset showing preservation of mitochondria, nucleus, nuclear envelope, and large (white boxes) and small (arrows) lysosomes (scale bar: 2 µm). (g) hiPSC-derived cortical neurons showing preservation of mitoGFP in a 200 nm thick easyIRF section, and (h) the matching ultrastructure (scale bar: 20 µm). Insets show that the pattern of mitoGFP in the LM images matches the pattern of mitochondria in the EM image (scale bar: 5 µm).

Article Snippet: Human cervical cancer epithelial (HeLa) cells were obtained from the American Type Culture Collection (ATCC, CCL-2).

Techniques: Preserving, Derivative Assay

Journal: bioRxiv

Article Title: VP-CLEM-Kit: An accessible pipeline for visual proteomics using super resolution volume correlative light and electron microscopy (SR-vCLEM)

doi: 10.1101/2025.04.08.647770

Figure Lengend Snippet: (a) System diagram of the tomoSTORM open-hardware microscope based on the openFrame modular system, showing the hardware components used to acquire RF, WF and SMLM array tomography images. The system can switch between imaging modalities by manually switching the reflection and multi-line fluorescence dichroic cubes using the dichroic switcher layer. There is a manual 4-objective turret with 10x (Amscope, 10x/0.3 NA) and 100x (Amscope, 100x,/1.3 NA) objectives. The TEC-cooled CMOS camera is a CellCam Kikker (Cairn Research Ltd, CellCam Kikker). The system uses a XY stage (Märzhäuser Wetzlar GmbH, SCAN IM 130 x 85) and a Z-stepper motor (Cairn Research Ltd, Z-Act). The openFrame has an openAF autofocus unit to maintain focus during SMLM acquisitions. The excitation source is a 4-line multimode laserbank (Cairn Research Ltd, Laserbank) which is fibre coupled into the critical illuminator. (b) A photo showing the tomoSTORM light microscope with the individual hardware components labelled. (c) The easyIRF resin block was sectioned and section ribbons retrieved onto an ITO-coated coverslip. The coverslip was attached to a slide-sized metal mount and placed into the slide-holding insert on the xy stage for imaging. (d) RF and (e) WF tiled images were acquired with the 10x objective from a ribbon of easyIRF sections containing Hela cells expressing mitoGFP. Insets (white boxes) show sections 4-6 of the ribbon. Tiled WF images were recorded using the 100x/ 1.3 NA objective from section 4. (f) SMLM of mitoGFP in HeLa cells in an easyIRF section. Left: Multiple frames of a single FOV show blinking in sequential frames. Middle: A single frame of the raw microscope acquisition alongside WF, compared to Right: the final SMLM reconstruction in section 6 of the same sample, highlighting the resolution improvement achieved with SMLM.

Article Snippet: Human cervical cancer epithelial (HeLa) cells were obtained from the American Type Culture Collection (ATCC, CCL-2).

Techniques: Microscopy, Tomography, Imaging, Fluorescence, Light Microscopy, Blocking Assay, Expressing