Journal: Nanomaterials

Article Title: Anticancer Potential of L-Histidine-Capped Silver Nanoparticles against Human Cervical Cancer Cells (SiHA)

doi: 10.3390/nano11113154

Figure Lengend Snippet: ( I ) Antiproliferative effect of L-HAgNPs against SiHa cells; ( II ) phase-contrast microscopic images to assess structural changes in cells treated with L-HAgNPs; ( III ) fluorescent images illustrating intracellular ROS level; ( IV ) bar graph depicting quantitative measurement of green fluorescent intensity proportional to ROS level. * p < 0.05 denotes statistical significance between control vs. treated groups.

Article Snippet: SiHa cells were purchased from National Centre for Cell Science (NCCS), Pune, India.

Techniques: Control

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: Frontiers in Oncology

Article Title: Mechanisms and Applications of the Anti-cancer Effect of Pharmacological Ascorbic Acid in Cervical Cancer Cells

doi: 10.3389/fonc.2020.01483

Figure Lengend Snippet: Effects of L-AA on other human cervical cancer cell lines. C33A (A) and SiHa (B) cells (2 × 10 5 cells/well) were incubated for 24 h with the indicated concentrations of L-AA. (C,D) HeLa cells (2 × 10 5 cells/well) were incubated for 24 h with the indicated concentrations of (C) sodium ascorbate or (D) indicated reagents. Cell lysates were subjected to Western blot analysis using antibodies against Nrf2, p62, e-IF2α, and p-e-IF2α. ACTN was the protein loading control. The results are representative of three independent experiments. Protein bands were quantified through pixel density scanning and evaluated using ImageJ, version 1.44a ( http://imagej.nih.gov/ij/ ). The fold was normalized to the internal control protein (ACTN).

Article Snippet: HeLa cervical carcinoma (ID 60005), C33A (ID 60554), and SiHa (ID 60528) cells were purchased from Bioresource Collection and Research Center, Taiwan, Republic of China.

Techniques: Incubation, Western Blot, Control

Journal: PLoS ONE

Article Title: Inactivation of SLIT2-ROBO1/2 Pathway in Premalignant Lesions of Uterine Cervix: Clinical and Prognostic Significances

doi: 10.1371/journal.pone.0038342

Figure Lengend Snippet: Bars represented the gene expression normalized to β2- microglobulin and relative to a pool of normal cervical tissues, using the -ddCt method. Bars 22 and 23 represent the SiHa and HeLa cell lines, respectively. Analysis of promoter methylation of ROBO1/2 and SLIT2 by MSRA and validation by MSP. ( C1 ) Schematic representation of promoter regions of candidate genes revealing distribution of Hpa II/ Msp I (CCGG: arrowhead) restriction sites. →: location of methylation primers. +1 : transcription start site. ( C2 ) Representative tumor samples (#3920T, #3229T, #4025T) showing methylated status at different genes, normal cervical tissue were unmethylated. M and H : Msp I and Hpa II digested DNA respectively. U: Undigested DNA. K1 and K2 : Controls for DNA digestion and integrity respectively, T: Tumor DNA, N: DNA from corresponding normal tissue. ( C3 ) Representative tumor samples showing methylation status of candidate genes by MSP. U: amplicon obtained with primer for bisulphite modified unmethylated DNA, M: amplicon obtained with primer for bisulphite modified methylated DNA, T: Tumor DNA, N: DNA from corresponding normal tissue. ( D ) Reactivation of RNA expression of ROBO1 and the ncRNAs in SiHa and HeLa cells. Reactivated RNA expression of the ligand-receptor genes in SiHa ( E ) and HeLa ( F ) cell lines in presence of 5 µM, 10 µM and 20 µM 5-aza dC. The bars represent increased gene expression in 5-aza dC treated cells compared to untreated control.

Article Snippet: Similar trend (except for ROBO1) was observed for the CACX cell lines SiHa and HeLa, in the following order: SiHa: SLIT2 (38 folds) > ROBO2 (29 folds), HeLa: ROBO2 (80 folds) > SLIT2 (2.8 folds) ( ).

Techniques: Gene Expression, Methylation, Biomarker Discovery, Amplification, Modification, RNA Expression, Control

Journal: PLoS ONE

Article Title: Inactivation of SLIT2-ROBO1/2 Pathway in Premalignant Lesions of Uterine Cervix: Clinical and Prognostic Significances

doi: 10.1371/journal.pone.0038342

Figure Lengend Snippet: (i) LOH: loss of heterozygosity, (ii) MA-1: microsatellite size alteration of one allele. (iii) LOH + MA: loss of one allele and microsatellite size alteration of the other. (iv) Hemizygous (HE) deletion of ROBO2 locus as shown by D3S2515. (v) & (vi) HE deletion as shown by exonic markers (EM) from ROBO1 and ROBO2 respectively, SST used as control. The sample numbers and marker loci are indicated above and below the figure respectively. →: allelic loss, “*”: allelic size alteration. Deletion of ROBO1/2 and SLIT2 analyzed by microsatellite and exonic markers in ( B ) CIN and ( C ) CACX. T: Tumor DNA, N: DNA from normal cervix/PBL. ( D ) Pattern of deletion and methylation of ROBO1/2 and SLIT2 during disease progression. Asterisk denotes statistical significance (P<0.05). ( E ) Overall alteration patterns of the individual genes, SLIT2-ROBO1 and SLIT2-ROBO2 ligand-receptor pairs, during disease progression. Asterisk denotes statistical significance (P<0.05).

Article Snippet: Similar trend (except for ROBO1) was observed for the CACX cell lines SiHa and HeLa, in the following order: SiHa: SLIT2 (38 folds) > ROBO2 (29 folds), HeLa: ROBO2 (80 folds) > SLIT2 (2.8 folds) ( ).

Techniques: Control, Marker, Methylation, Biomarker Discovery

Journal: PLoS ONE

Article Title: Inactivation of SLIT2-ROBO1/2 Pathway in Premalignant Lesions of Uterine Cervix: Clinical and Prognostic Significances

doi: 10.1371/journal.pone.0038342

Figure Lengend Snippet: Correlation between deletion/methylation and reduced expression (RNA/protein) of ROBO1/2 and SLIT2 in CIN/CACX.

Article Snippet: Similar trend (except for ROBO1) was observed for the CACX cell lines SiHa and HeLa, in the following order: SiHa: SLIT2 (38 folds) > ROBO2 (29 folds), HeLa: ROBO2 (80 folds) > SLIT2 (2.8 folds) ( ).

Techniques: Expressing

Journal: PLoS ONE

Article Title: Inactivation of SLIT2-ROBO1/2 Pathway in Premalignant Lesions of Uterine Cervix: Clinical and Prognostic Significances

doi: 10.1371/journal.pone.0038342

Figure Lengend Snippet: ( A ) In normal cervical epithelium the basal and parabasal layer stained intensely for all the three proteins, whereas the intensity and frequency of stained cells reduced with further differentiation in the spinous layer. ( B ) In primary CACX expression pattern of these proteins were concordant with respective molecular alterations. ( C ) In SiHa cells ROBO1 and ROBO2 were membrane localized, whereas SLIT2 was present mostly in the cytoplasm. T: primary CACX sample; scale bars for both 20X and 40X is 50 µm; original magnifications are indicated in parenthesis. Magnification of panel C is 40X.

Article Snippet: Similar trend (except for ROBO1) was observed for the CACX cell lines SiHa and HeLa, in the following order: SiHa: SLIT2 (38 folds) > ROBO2 (29 folds), HeLa: ROBO2 (80 folds) > SLIT2 (2.8 folds) ( ).

Techniques: Staining, Expressing, Membrane

Journal: PLoS ONE

Article Title: Inactivation of SLIT2-ROBO1/2 Pathway in Premalignant Lesions of Uterine Cervix: Clinical and Prognostic Significances

doi: 10.1371/journal.pone.0038342

Figure Lengend Snippet: Alterations of ( A ) SLIT2 and/or ROBO1 and ( B ) SLIT2 and/or ROBO2 ligand-receptor pairs were significantly associated with poor patient outcome [OS]. ( C ) Representation of Cox Multivariate analyses of genetic, clinical and etiological parameters in predicting outcome of CACX patients. N: total number of samples and P<0.05 denotes statistical significance.

Article Snippet: Similar trend (except for ROBO1) was observed for the CACX cell lines SiHa and HeLa, in the following order: SiHa: SLIT2 (38 folds) > ROBO2 (29 folds), HeLa: ROBO2 (80 folds) > SLIT2 (2.8 folds) ( ).

Techniques: