Journal: International journal of cancer

Article Title: Autocrine/paracrine erythropoietin signalling promotes JAK/STAT-dependent proliferation of human cervical cancer cells.

doi: 10.1002/ijc.25935

Figure Lengend Snippet: Figure 3. Exogenous Epo induces proliferation of cervical cancer cells. SiHa, HeLa and C33 cells were treated with increasing

Article Snippet: Cervical cancer-derived HeLa, SiHa and C33 cells (American Type Culture Collection, Manassas VA) were maintained in Dulbecco’s modified Eagle’s medium (DMEM; Invitro, Mexico DF, Mexico) supplemented with 2% heat-inactivated foetal bovine serum (FBS) (Invitrogen, Carlsbad, CA).

Techniques:

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) RT-qPCR for the expression of CCHCR1 and TCF19 in HeLa cells transfected with E2F activators (E2F1, E2F2 or E2F3). pEGFP-N1 was the empty vector control. Gene expression level was normalized to the expression level of RPL13A . Results were mean ± SD of three replicates. Statistically significant differences of the gene expression levels were compared to empty vector control and were analyzed by paired t-test (** = P ≤ 0.01). ( b ) Down-regulation of CCHCR1 and TCF19 mRNA by E2F1 knockdown. Cells were transfected with siE2F1 and were harvested for the time indicated (time post-transfection). mRNA expressions (normalized to RPL13A gene expression) were analyzed by RT-qPCR. Results were mean ± SD of three replicates. Statistically significant differences of the gene expression levels were compared to 0 h post-siE2F1 transfection and were analyzed by paired t-test (*** = P ≤ 0.001).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Quantitative RT-PCR, Expressing, Transfection, Plasmid Preparation, Control, Gene Expression, Knockdown

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were synchronized and arrested in G1/S transition by a double thymidine block. The cells were released and collected at different time points, stained with propidium iodide (PI). The percentage of cells in different cell cycle phases in each time point (G1/S, S, G2/M, and G1) were analyzed by flow cytometry. ( b ) Percentage of cells in different cell cycles after double thymidine block and release. (C) Relative expression of CCHCR1 and TCF19 in different cell cycle phases. CCHCR1 and TCF19 gene expression were normalized to RPL13A gene expression. The time points after release from the double thymidine block were indicated for different cell cycle phases; i.e. 0 h: G1/S, 2 h: S, 8 h: G2/M and 14 h: G1. Results were mean ± SD of three replicates. Statistically significant differences of the gene expressions were compared to 0 h after double thymidine release (G1/S transition) and were analyzed by paired t-test (** = P ≤ 0.01 and *** = P ≤ 0.001)

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Blocking Assay, Staining, Flow Cytometry, Expressing, Gene Expression

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were transfected with the corresponding siRNAs. Non-viable cells were counted using trypan blue exclusion assay. siCTL was Silencer select negative control #1 (Ambion, Life Technologies, USA). Results are mean ± SD of five replicates. Statistically significant differences of the cell counts were compared to the untransfected samples in the corresponding time point, and were analyzed by paired t-test (* = ≤0.05 and ** = P ≤ 0.01). ( b ) Up-regulation of p21 mRNA was observed after knockdown of CCHCR1 . siRNAs were transfected into HeLa cells and were collected for analysis 4 days post-transfection. Gene expressions (normalized to RPL13A gene expression) were analyzed by RT-qPCR. Results are mean ± SD of three replicates. Statistically significant differences of the gene expressions were compared to untransfected sample and were analyzed by paired t-test (* = ≤0.05, ** = P ≤ 0.01 and *** = P ≤ 0.001).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Transfection, Trypan Blue Exclusion Assay, Negative Control, Knockdown, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Transcriptional Regulation of the P-bodies Associated Psoriasis Candidate Gene CCHCR1

doi: 10.1101/2023.04.08.535951

Figure Lengend Snippet: ( a ) HeLa cells were treated with 0.1% DMSO (vehicle control) or 2 μM CPT for 24 hours. The percentage of cells in different cell cycle phases were analyzed by flow cytometry. ( b ) Relative expression of CCHCR1 and TCF19 (normalized to RPL13A gene expression) in HeLa cells after the treatment with CPT for 24 hours. Results are mean ± SD of three replicates. Statistically significant differences of the gene expression level were compared to 0.1% DMSO treatment and were analyzed by paired t-test (* = ≤0.05 and ** = P ≤ 0.01).

Article Snippet: Human cervical cancer HeLa cells (ATCC ® CRM-CCL-2 TM ) were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and maintained at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Control, Flow Cytometry, Expressing, Gene Expression

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 is highly expressed in CC tissues and cells. (A) Heatmap of differentially expressed circRNAs in CC tissues and normal tissues according to the online data set (GSE102686). (B) Expression level of hsa_circ_0101119 was detected via reverse transcription-quantitative PCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and normal human cervical epithelial cell line, HcerEpic. **P<0.01 vs. HcerEpic cells. (C) Expression level of hsa_circ_0101119 in CC tissues and normal tissues, according to the online data set (GSE102686). circRNA/circ, circular RNA.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: hsa_circ_0101119 facilitates the progression of cervical cancer via an interaction with EIF4A3 to inhibit TCEAL6 expression

doi: 10.3892/mmr.2021.12293

Figure Lengend Snippet: hsa_circ_0101119 recruits EIF4A3 to inhibit TCEAL6 expression in CC. (A) Bioinformatics was used to predict the interaction probabilities of the RNA-binding protein EIF4A3 with hsa_circ_0101119. Predictions with probabilities >0.5 were considered ‘positive’, suggesting that the corresponding RNA and protein are likely to interact. (B) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated hsa_circ_0101119 in SiHa and HeLa cell lysates. (C) Pull down assay indicated that biotin-labeled hsa_circ_0101119 interacted with EIF4A3. (D) Bioinformatics was used to predict the interaction probabilities of EIF4A3 with TCEAL6. (E) RIP assay using anti-EIF4A3 showed that EIF4A3 precipitated TCEAL6 in SiHa and HeLa cell lysates. (F) Expression levels of EIF4A3 and TCEAL6 were detected via RT-qPCR in CC cell lines (C-33A, SiHa, CaSki and HeLa) and a normal human cervical epithelial cell line, HcerEpic. (G) Expression levels of EIF4A3 and TCEAL6 in CC tissues and normal tissues, according to the analysis of TCGA. (H) Correlation between EIF4A3 and TCEAL6 in CC samples from TCGA. (I) After transfection with sh-EIF4A3, RT-qPCR was used to detect EIF4A3 expression in SiHa and HeLa cells. (J) After transfection with sh-EIF4A3, western blotting was performed to detect the expression level of TCEAL6 in SiHa and HeLa cells. (K) After co-transfection with si-hsa_circ_0101119 and sh-EIF4A3, western blotting was performed to measure the expression level of TCEAL6 in SiHa and HeLa cells. (L) A proposed model whereby hsa_circ_0101119 sequesters EIF4A3 away from TCEAL6 mRNA, in turn suppressing TCEAL6 mRNA translation. **P<0.01 vs. IgG group (B and E); *P<0.05, **P<0.01 vs. HcerEpic cells group (F); *P<0.05 vs. normal tissues group (G); **P<0.01, vs. sh-NC group (I and J); **P<0.01 vs. sh-NC group, ## P<0.01, vs. si-hsa_circ group. (K) RIP, RNA immunoprecipitation; RT-qPCR, reverse transcription-quantitative PCR; TCGA, The Cancer Genome Atlas; sh, short hairpin RNA; NC, negative control; si, small interfering RNA; circ, circular RNA; EIF4A3, eukaryotic initiation factor 4A-3; TCEAL6, transcription elongation factor A-like 6; T, tumor; N, normal; CC, cervical cancer.

Article Snippet: The four human CC cell lines (C-33A, SiHa, CaSki and HeLa) and the normal human cervical epithelial cell line, HcerEpic, were supplied by VCANBIO Cell & Gene Engineering Co., Ltd. All the cells were cultured in RPMI 1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% (v/v) FBS (Invitrogen; Thermo Fisher Scientific, Inc.) and 1% penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.), and cultured at 37°C in a humidified 5% CO 2 incubator.

Techniques: Expressing, RNA Binding Assay, Pull Down Assay, Labeling, Quantitative RT-PCR, Transfection, Western Blot, Cotransfection, Immunoprecipitation, Real-time Polymerase Chain Reaction, shRNA, Negative Control, Small Interfering RNA

Journal: Biomolecules and Biomedicine

Article Title: Role of Fascin-1 in cervical cancer metastasis via Wnt/β-catenin pathway activation

doi: 10.17305/bb.2025.12114

Figure Lengend Snippet: Impact of Fascin-1 on in vitro growth of cervical cancer cells. (A) Assessment of Fascin-1 levels in Hela and C33A cells through Western blot, post-infection with lentiviruses carrying either FSCN1 -targeting shRNAs or control shRNAs; (B) RT-PCR analysis of FSCN1 mRNA levels when knockdown FASCIN; (C) Evaluation of cell proliferation in Hela and C33A cells with FSCN1 or control shRNA expression, using MTT assays; (D) Growth of Hela and C33A cells expressing respective shRNAs, cultured for 10 days and stained with crystal violet; colony counts were recorded; (E) Western blot analysis of cell lysates from Hela and C33A cells stably expressing FLAG- FSCN1 empty vector; (F) RT-PCR analysis of FSCN1 mRNA levels when overexpressed FSCN1 ; (G) Proliferation analysis via MTT assays in cells with either control or FSCN1 overexpression; (H) Analysis of colony formation in cells with indicated treatments; (I) Cell cycle experiments in indicated cells. Data for – and – are presented as mean ± SD, n ═ 3; *** P < 0.001, ** P < 0.01, * P < 0.05 (Student’s t -test). shRNA: Short hairpin RNA.

Article Snippet: The Hela and C33A human cervical cell lines were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in RPMI 1640 and MEM media, respectively, both supplied by Meilunbio (Dalian, China).

Techniques: In Vitro, Western Blot, Infection, Control, Reverse Transcription Polymerase Chain Reaction, Knockdown, shRNA, Expressing, Cell Culture, Staining, Stable Transfection, Plasmid Preparation, Over Expression

Journal: Biomolecules and Biomedicine

Article Title: Role of Fascin-1 in cervical cancer metastasis via Wnt/β-catenin pathway activation

doi: 10.17305/bb.2025.12114

Figure Lengend Snippet: Enhancement of cervical cell migration by Fascin-1 in vitro . (A and B) Migration and invasion capacities of Hela and C33A cells, with either FSCN1 shRNAs or control shRNAs, were assessed through respective assays; (C and D) Cells overexpressing either empty vectors or FLAG-tagged FSCN1 were analyzed for migration and invasion abilities. Scale bars represent 100 µm. Statistical data are presented as mean ± SD ( n ═ 3); *** P < 0.001, ** P < 0.01, * P < 0.05 (Student’s t -test).

Article Snippet: The Hela and C33A human cervical cell lines were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in RPMI 1640 and MEM media, respectively, both supplied by Meilunbio (Dalian, China).

Techniques: Migration, In Vitro, Control

Journal: Biomolecules and Biomedicine

Article Title: Role of Fascin-1 in cervical cancer metastasis via Wnt/β-catenin pathway activation

doi: 10.17305/bb.2025.12114

Figure Lengend Snippet: Downregulation of Fascin-1 impairs Wnt/β-catenin signaling. (A) GSEA enrichment plot of Wnt signaling pathway using RNA-seq data generated from Hela cells; (B) Heatmap of genes involved in Wnt signaling pathway; (C) mRNA was isolated from Hela and C33A cells expressing specific shRNAs, and quantitative real-time RT-PCR assays were conducted to measure expression levels; (D) RT-PCR analysis of FSCN1 , CTNNB1 and c-Myc mRNA levels in the indicated cells; (E and F) Protein lysates from indicated Hela and C33A cells, were analyzed by Western blot using relevant antibodies; (G) pTop-Flash/pFop-Flash reporter vectors were used to evaluate the transcriptional activity of the Wnt/β-catenin signaling pathway. Each bar in the graph indicates the average ± SD from three independent experiments, *** P < 0.001 compared to control (Student’s t -test). GSEA: Gene set enrichment analysis.

Article Snippet: The Hela and C33A human cervical cell lines were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China) and cultured in RPMI 1640 and MEM media, respectively, both supplied by Meilunbio (Dalian, China).

Techniques: RNA Sequencing, Generated, Isolation, Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Western Blot, Activity Assay, Control