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:

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: PLoS ONE

Article Title: Induction of apoptosis by pinostrobin in human cervical cancer cells: Possible mechanism of action

doi: 10.1371/journal.pone.0191523

Figure Lengend Snippet: Cytotoxicity % (CT) was assessed in cervical cell lines (A) HeLa, (B) Ca Ski, (C) SiHa cells on different concentrations of P N treatments as determined by MTT reduction assay at different incubation period. The bar graphs represent the percentage of cytotoxicity of P N in the cells. Cytotoxicity is shown as mean ± SD derived from at least three separate experiments in triplicate wells. Ordinary two-way ANOVA (multiple comparisons) was performed to calculate the statistical difference ( p≤0 . 05 ) among all treated groups as compared to vehicle treated group. * represents p ≤0.05, ∞ p ≤0.01, $ p ≤0.001 and # p ≤0.0001.

Article Snippet: HeLa (human adeno cervical carcinoma cells), Ca Ski (human epidermoid cervical carcinoma cells), SiHa (Grade II, squamous cervical carcinoma cell) and HEK293 (Human embryonic kidney cells) cell lines were procured from culture collection at National Centre for Cell Science (NCCS, Pune) India.

Techniques: MTT Reduction Assay, Incubation, Derivative Assay