Journal: OncoTargets and therapy

Article Title: Gemcitabine and carboplatin demonstrate synergistic cytotoxicity in cervical cancer cells by inhibiting DNA synthesis and increasing cell apoptosis

doi: 10.2147/OTT.S54217

Figure Lengend Snippet: Carboplatin reduced cell viability and induced DNA damage in cervical cancer cells. ( A ) SiHa and CaSki cells were treated with the indicated concentrations of carboplatin for 72 hours, and cell viability was measured by Cell Counting Kit-8 viability assay. ( B ) Carboplatin induced DNA damage in SiHa cells. SiHa cells were exposed to 20, 40, and 80 μmol/L carboplatin for 12 hours, respectively. Immunofluorescence analysis was used to detect nuclear γ-H2AX foci formation with anti-H2AX antibody (red, fluorescein isothiocyanate). Nuclei were counterstained with DAPI (blue). ( C ) γ-H2AX-positive cells were counted under a fluorescent microscope. We calculated more than 1,000 cells for each well. Quantitative data are represented the mean ± standard deviation of three individual experiments. Abbreviation: DAPI, 4′,6-diamidino-2-phenylindole.

Article Snippet: SiHa and CaSki human cervical cancer cell lines (American Type Culture Collection, Manassas, VA, USA) were maintained in RPMI-1640 medium (Gibco, Carlsbad CA, USA) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and 100 U/mL penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Cell Counting, Viability Assay, Immunofluorescence, Microscopy, Standard Deviation

Journal: OncoTargets and therapy

Article Title: Gemcitabine and carboplatin demonstrate synergistic cytotoxicity in cervical cancer cells by inhibiting DNA synthesis and increasing cell apoptosis

doi: 10.2147/OTT.S54217

Figure Lengend Snippet: Synergistic cytotoxicity of gemcitabine combined with carboplatin in cervical cancer cell lines. ( A ) SiHa and CaSki cells were seeded into 96-well plates and treated with gemcitabine or/and carboplatin at the indicated concentrations. After 72 hours, cell viability was measured using the Cell Counting Kit-8 viability assay. The data shown represent the mean ± standard deviation (n=3). ( B ) The synergistic effect of gemcitabine combined with carboplatin was quantitatively analyzed with a Cl and expressed as log 10 (CI) versus fractional effect Where calculable, 95% confidence intervals are shown. Abbreviations: DAPI, 4′,6-diamidino-2-phenylindole; GEM, gemcitabine; CBDCA, carboplatin; CI, combination index; SD, standard deviation.

Article Snippet: SiHa and CaSki human cervical cancer cell lines (American Type Culture Collection, Manassas, VA, USA) were maintained in RPMI-1640 medium (Gibco, Carlsbad CA, USA) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and 100 U/mL penicillin-streptomycin at 37°C in a humidified atmosphere of 5% CO 2 .

Techniques: Cell Counting, Viability Assay, Standard Deviation

Journal: PLoS ONE

Article Title: Over-Expressed miR-224 Promotes the Progression of Cervical Cancer via Targeting RASSF8

doi: 10.1371/journal.pone.0162378

Figure Lengend Snippet: (A) Wound healing assay was implemented in cervical cancer cell line SiHa and CaSki after transfected with 50 nM of miR-224 mimic or negative control (miR-NC). The wound healing was measured at the time points as indicated. Bars represented the average percentage of wound healing ±s.d. ** P<0.01. (B) Cell invasion capabilities were measured in SiHa and CaSki cells with transwell chambers, as described in materials and methods. Photos were representative fields of invasive cells on the membrane with a 200-fold magnification. Bar graphs represented the average number of cells per field on the underside of the membrane ±s.d. ** P<0.01.

Article Snippet: SiHa and CaSki cell lines, purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cell bank of Chinese Academy of Science in Shanghai respectively, were cultured at 37°C and 5% CO2 in a humidified incubator in Dulbecco’s modified Eagle’s medium (DMEM) and 1640 medium separately.

Techniques: Wound Healing Assay, Transfection, Negative Control, Membrane

Journal: PLoS ONE

Article Title: Over-Expressed miR-224 Promotes the Progression of Cervical Cancer via Targeting RASSF8

doi: 10.1371/journal.pone.0162378

Figure Lengend Snippet: (A,B) At 72h after transfected with 100 nM of miRNA mimic and negative control, the endogenous protein levels of RASSF8 in SiHa and CaSki cells were measured by western blot. Bars indicated the relative protein levels that were normalized to GAPDH. Data were presented as mean±s.d. (n¼3) *P<0.05, **P<0.01. (C) Putative miR-224-binding site in the RASSF83'-UTR mutation was generated by mutating 3nt that is recognized by miR-224. Either wild-type (WT) or mutant RASSF8 3'-UTR was subcloned into the dual-luciferase reporter vector. (D) The luciferase reporter vector containing wild (wild type) RASSF8 3’-UTR or mutant RASSF8 3’-UTR was cotransfected into SiHa cells with miR-224 mimic or miRNA negative control. (E) Firefly luciferase activities were determined at 48h postransfection and normalized to Renilla luciferase. Each column represented the mean±s.d. of three independent experiments. **P <0.01.

Article Snippet: SiHa and CaSki cell lines, purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cell bank of Chinese Academy of Science in Shanghai respectively, were cultured at 37°C and 5% CO2 in a humidified incubator in Dulbecco’s modified Eagle’s medium (DMEM) and 1640 medium separately.

Techniques: Transfection, Negative Control, Western Blot, Binding Assay, Mutagenesis, Generated, Luciferase, Plasmid Preparation

Journal: PLoS ONE

Article Title: Over-Expressed miR-224 Promotes the Progression of Cervical Cancer via Targeting RASSF8

doi: 10.1371/journal.pone.0162378

Figure Lengend Snippet: (A) Specific siRNA targeting RASSF8 suppressed RASSF8 protein expression. SiHa and CaSki cells were transfected with 50 nmol of siRNA RASSF8 or siRNA-negative control (siR-NC) and the transfection efficiency were assessed. RASSF8 protein expression levels were determined by western blot analysis. GAPDH was served as the internal control. (B)Representative images of wound healing were taken at 0h, 24h, and 48h after the wound scratched. Data are presented as mean±s.d. *P<0.05, **P<0.01. (C) The number of invaded cells following RASSF8 knockdown using specific siRNA targeting RASSF8(siR-RASSF8-a) versus negative control (siR-NC) were shown by stained with crystal violet. Bars indicated the invasion capability compared with that of negative control ± s.d*P<0.05, **P<0.01.

Article Snippet: SiHa and CaSki cell lines, purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cell bank of Chinese Academy of Science in Shanghai respectively, were cultured at 37°C and 5% CO2 in a humidified incubator in Dulbecco’s modified Eagle’s medium (DMEM) and 1640 medium separately.

Techniques: Expressing, Transfection, Negative Control, Western Blot, Control, Knockdown, Staining