human cervical cancer cell line c33a  (ATCC)

Journal: Cell Death & Disease

Article Title: Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates

doi: 10.1038/s41419-024-07073-y

Figure Lengend Snippet: A The expression of ZDHHC3 protein in cancers based on the Human Protein Atlas: ZDHHC3 expression levels are evaluated according to the immunohistochemistry staining on cancer tissue samples of the Human Protein Atlas database. The X axis represents the cancer type. Pan-cancer includes 20 different cancer types. BRCA, breast cancer. CESC, cervical cancer. SKCM, skin cancer and melanoma. HNSC, head and neck cancer. The Y axis represents the percentage of patients with specific expression pattern of ZDHHC3. High, high expression. Low, low expression. None, no expression. B The correlation between PDL1 (CD274) and PD1(PDCD1), ZDHHC1, ZDHHC2, or ZDHHC3 expression in TCGA cancer types. TIMER2.0 Gene_Corr Module is used to evaluate the expression between two genes in different cancer types of TCGA database. The heatmap lists the purity-adjusted partial spearman’s rho (Cor) value as the degree of the correlation between two genes and the number (n) of patient cases in each cancer typer. C Western blot analysis was utilized to examine the expression of PD-L1 protein in MDA-MB-231 and C33A cells following the knockdown of DHHC1, DHHC2, and DHHC3. D A schematic representation of the structural design of cp-PCC. E Western blot analysis was conducted to assess the expression of PD-L1 protein in four cell lines (MDA-MB-231, C33A, FaDu, A375) following a 6-hour incubation with 10 µM PCC16/17/18, each corresponding to CRBN/VHL/IAP-based cp-PCC, respectively. Quantitative data are presented as mean ± standard error. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test ( n = 3): * P < 0.05, ** P < 0.01, ***P < 0.001.

Article Snippet: Human cervical cancer cell line C33A, mouse mammary carcinoma cell line 4T1, and mouse cervical cancer cell line U14 were purchased from ATCC.

Techniques: Expressing, Immunohistochemistry, Staining, Western Blot, Knockdown, Incubation

Journal: Cell Death & Disease

Article Title: Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates

doi: 10.1038/s41419-024-07073-y

Figure Lengend Snippet: A – C Western blot analysis was utilized to examine the expression of DHHC3/PD-L1 proteins in C33A cells following 6 h of treatment with varying concentrations of PCC16/17/18. D Comparative analysis of the DC50 values for PCC16/17/18. E – G Western blot analysis to assess the expression of DHHC3/PD-L1 proteins in C33A cells post-incubation with specific concentrations of PCC16/17/18 for different time periods. H Confocal microscopy was employed to observe the fluorescence intensity of PCC16/17/18 entering the cells at various time points. Quantitative data are presented as mean ± standard error. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test ( n = 3): * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human cervical cancer cell line C33A, mouse mammary carcinoma cell line 4T1, and mouse cervical cancer cell line U14 were purchased from ATCC.

Techniques: Western Blot, Expressing, Incubation, Confocal Microscopy, Fluorescence

Journal: Cell Death & Disease

Article Title: Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates

doi: 10.1038/s41419-024-07073-y

Figure Lengend Snippet: A Confocal observation of DHHC3 binding with drug PCC16. Scale bar, 1 µM. B C33A cell lysates treated at different temperatures after incubation with 0.5 µM PCC16, with Western blot analysis of DHHC3 protein expression; C CETSA melting curve plotted. D Flow cytometry analysis of fluorescent cells after treatment with Rhodamine-labeled PCC16 at different concentration gradients; E C33A cells treated with PCC16 over time and concentration gradients, with cell toxicity of PCC16 measured by MTT assay.

Article Snippet: Human cervical cancer cell line C33A, mouse mammary carcinoma cell line 4T1, and mouse cervical cancer cell line U14 were purchased from ATCC.

Techniques: Binding Assay, Incubation, Western Blot, Expressing, Flow Cytometry, Labeling, Concentration Assay, MTT Assay

Journal: Cell Death & Disease

Article Title: Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates

doi: 10.1038/s41419-024-07073-y

Figure Lengend Snippet: A C33A cells pre-treated with/without MG132 for 4 h and then treated with PCC16/17/18, with Western blot analysis to detect PD-L1 protein levels; B Bar graph representing the quantification of grayscale values; C Confocal microscopy observation of the immunofluorescence intensity of PD-L1 protein under PCC16/17/18 ± MG132 conditions. Scale bar, 10 μM. Quantitative data expressed as mean ± standard error; analyzed by one-way ANOVA with Tukey’s post hoc test ( n = 3): * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human cervical cancer cell line C33A, mouse mammary carcinoma cell line 4T1, and mouse cervical cancer cell line U14 were purchased from ATCC.

Techniques: Western Blot, Confocal Microscopy, Immunofluorescence

Journal: Cell Death & Disease

Article Title: Treating ICB-resistant cancer by inhibiting PD-L1 via DHHC3 degradation induced by cell penetrating peptide-induced chimera conjugates

doi: 10.1038/s41419-024-07073-y

Figure Lengend Snippet: A Western blot analysis of the effects of PCC series drugs and PD-L1 monoclonal antibody on PD-L1 protein levels in C33A cells; B TUNEL assay to assess apoptosis in cervical cancer cells treated with cisplatin and PCC16; C Ki67 assay for evaluating the proliferation of cervical cancer cells treated with cisplatin and PCC16; D Plate cloning experiment analyzing the proliferation of cervical cancer cells treated with cisplatin and PCC16; E In a C33A-T cell co-culture model, ELISA was used to measure IFN-γ and TNF-α expression levels after treatment with different concentrations of PCC16; F Hoechest33528 staining in the co-culture system to observe apoptosis of C33A cells after treatment with different concentrations of PCC16. Quantitative data are expressed as mean ± standard error; analyzed by one-way ANOVA with Tukey’s post hoc test ( n = 3): * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human cervical cancer cell line C33A, mouse mammary carcinoma cell line 4T1, and mouse cervical cancer cell line U14 were purchased from ATCC.

Techniques: Western Blot, TUNEL Assay, Cloning, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Staining

Journal: Mediators of Inflammation

Article Title: Tumor Necrosis Factor- α Promotes the Tumorigenesis, Lymphangiogenesis, and Lymphatic Metastasis in Cervical Cancer via Activating VEGFC-Mediated AKT and ERK Pathways

doi: 10.1155/2023/5679966

Figure Lengend Snippet: TNF- α stimulates cervical cancer cells to secrete VEGFC and promotes HLEC viability. (a, b) Cervical cancer cells (HeLa and C33A) were stimulated with different concentrations of TNF- α (0, 5, or 10 ng/ml) for 48 h, and corresponding CM was collected. Next, the level of VEGFC in the corresponding CM was detected by ELISA. (c, d) HLECs were cultured in CM with or without MAZ51 treatment. HLEC viability was measured by CCK-8 assay. ∗∗ P < 0.01 compared with CM group; ## P < 0.01 compared with CM-TNF- α group, n = 3.

Article Snippet: HeLa and C33A human cervical cancer cell lines were purchased from the American Type Culture Collection and cultured in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Inc.) with 5% CO 2 at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, CCK-8 Assay

Journal: Mediators of Inflammation

Article Title: Tumor Necrosis Factor- α Promotes the Tumorigenesis, Lymphangiogenesis, and Lymphatic Metastasis in Cervical Cancer via Activating VEGFC-Mediated AKT and ERK Pathways

doi: 10.1155/2023/5679966

Figure Lengend Snippet: CM-TNF- α promotes the proliferation and angiogenesis of HLECs. Cervical cancer cells (HeLa and C33A) were stimulated with 10 ng/ml TNF- α for 48 h, and corresponding CM was collected. Next, HLECs were cultured in CM with or without MAZ51 treatment. (a, b) The proliferation of HLECs was measured by EdU staining assay. (c, d) The number of tube node of HLECs was observed using a microscope. ∗∗ P < 0.01 compared with CM group; ## P < 0.01 compared with CM-TNF- α group, n = 3.

Article Snippet: HeLa and C33A human cervical cancer cell lines were purchased from the American Type Culture Collection and cultured in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Inc.) with 5% CO 2 at 37°C.

Techniques: Cell Culture, Staining, Microscopy

Journal: Mediators of Inflammation

Article Title: Tumor Necrosis Factor- α Promotes the Tumorigenesis, Lymphangiogenesis, and Lymphatic Metastasis in Cervical Cancer via Activating VEGFC-Mediated AKT and ERK Pathways

doi: 10.1155/2023/5679966

Figure Lengend Snippet: CM-TNF- α increases the migration of HLECs. Cervical cancer cells (HeLa and C33A) were stimulated with 10 ng/ml TNF- α for 48 h, and corresponding CM was collected. Next, HLECs were cultured in CM with or without MAZ51 treatment. (a, b) The migration of HLECs was measured by wound healing assay. (c, d) The migration of HLECs was measured by transwell migration assay. ∗∗ P < 0.01 compared with CM group; ## P < 0.01 compared with CM-TNF- α group, n = 3.

Article Snippet: HeLa and C33A human cervical cancer cell lines were purchased from the American Type Culture Collection and cultured in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Inc.) with 5% CO 2 at 37°C.

Techniques: Migration, Cell Culture, Wound Healing Assay, Transwell Migration Assay

Journal: Mediators of Inflammation

Article Title: Tumor Necrosis Factor- α Promotes the Tumorigenesis, Lymphangiogenesis, and Lymphatic Metastasis in Cervical Cancer via Activating VEGFC-Mediated AKT and ERK Pathways

doi: 10.1155/2023/5679966

Figure Lengend Snippet: CM-TNF- α upregulates the expressions of p-AKT and p-ERK of HLECs. C33A cells were stimulated with 10 ng/ml TNF- α for 48 h, and the CM was collected. Next, HLECs were cultured in CM with or without MAZ51 treatment. (a–c) The levels of AKT, p-AKT, ERK, and p-ERK of HLECs were measured by western blot. ∗∗ P < 0.01 compared with CM group; ## P < 0.01 compared with CM-TNF- α group, n = 3.

Article Snippet: HeLa and C33A human cervical cancer cell lines were purchased from the American Type Culture Collection and cultured in DMEM (Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Thermo Fisher Scientific, Inc.) with 5% CO 2 at 37°C.

Techniques: Cell Culture, Western Blot