Journal: Frontiers in Cell and Developmental Biology

Article Title: Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca 2+ Flux Mediates Membrane Ruffling and Cell Survival

doi: 10.3389/fcell.2022.865056

Figure Lengend Snippet: Piezo1-mediated extracellular Ca 2+ influx induced by Yoda1 in various cell lines. (A) Viability of MCF-7 cells exposed to control [0.2% (v/v) DMSO] and Yoda1 (0.1–25 μM) for 24 h, measured using the WST-8 assay. The bar graphs describe mean values of cell viability with error bars indicating the standard error of the mean (S.E.M) ( n = 4, ** p < 0.01, student t -test). (B,D) Piezo1 expression in MCF-7 cells with or without control/Piezo1 shRNA as analysed by (B) RT-PCR and (D) western blotting. GAPDH was used as an internal control. (C,E) The bar graphs describe the cumulative densitometric analysis of (C) RT-PCR and (E) western blot bands normalized to the GAPDH. Data are presented as the means ± S.E.M ( n = 4, ** p < 0.01 and *** p < 0.001, student t -test). (F,H) Time-lapse FRET images of the Cytosolic-D3cpv in (F) MCF-7 cells, (H) HEK293A and HeLa cells introduced with or without exogenous Piezo1. MCF-7 cells were exposed to control [0.2% (v/v) DMSO] and Yoda1 (1 μM) dissolved in CO 2 -independent culture medium (Yoda1 group), Ca 2+ -free medium (Yoda1 + w/o Ca 2+ group), and Ca 2+ medium (Yoda1 + w/Ca 2+ group), respectively. (F) Yoda1 + Piezo1 KD group and (H) HEK293A and HeLa cells were incubated in CO 2 -independent culture medium. The color scale bars represent the range of the FRET/CFP emission ratio determined using the biosensor. Hot and cold colors indicate high and low Ca 2+ concentration, respectively. (G,I,J) The time courses represent the average of the normalized FRET/CFP emission ratio changes of cytosolic-D3cpv in (G) MCF-7, (I) HEK293A, and (J) HeLa cells. The lines are mean values of normalized emission ratios, with diluted colors indicating the S.E.M ( n = 7).

Article Snippet: The MCF-7, SiHa, and BeWo cell lines were purchased from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea), and HEK293A and HeLa cell lines were provided by Dr. Jihye Seong (Korea Institute of Science and Technology, Seoul, South Korea).

Techniques: Expressing, shRNA, Reverse Transcription Polymerase Chain Reaction, Western Blot, Incubation, Concentration Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Förster Resonance Energy Transfer-Based Single-Cell Imaging Reveals Piezo1-Induced Ca 2+ Flux Mediates Membrane Ruffling and Cell Survival

doi: 10.3389/fcell.2022.865056

Figure Lengend Snippet: Piezo1-dependent ER-stored Ca 2+ release in response to Yoda1. (A,D) Time-lapse FRET images of the ER Ca 2+ sensor in (A) MCF-7 and (D) HeLa cells. The cells were exposed to control [0.2% (v/v) DMSO] and Yoda1 (1 μM) dissolved in CO 2 -independent culture medium (Yoda1 group), Ca 2+ -free medium (Yoda1 + w/o Ca 2+ group), and Ca 2+ medium (Yoda1 + w/Ca 2+ group), respectively. The color scale bars represent the range of the FRET/CFP emission ratio determined using the biosensor. Hot and cold colors indicate high and low Ca 2+ concentration, respectively. (D) The red fluorescence protein images confirmed the expression of Piezo1-tdTomato. (B,E) The time courses represent the average of the normalized FRET/CFP emission ratio changes of the ER Ca 2+ sensor. The lines are mean values of the normalized emission ratios, with diluted colors indicating the S.E.M ( n = 7). (C,F) The bar graph describes the mean values of the normalized FRET/CFP emission ratios of the ER Ca 2+ sensor at the described time with error bars indicating the S.E.M ( n = 7, * p < 0.05 and ** p < 0.01, Student’s t -test).

Article Snippet: The MCF-7, SiHa, and BeWo cell lines were purchased from the Korean Cell Line Bank (KCLB; Seoul, Republic of Korea), and HEK293A and HeLa cell lines were provided by Dr. Jihye Seong (Korea Institute of Science and Technology, Seoul, South Korea).

Techniques: Concentration Assay, Fluorescence, Expressing

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: Journal of Translational Medicine

Article Title: RGS1 and related genes as potential targets for immunotherapy in cervical cancer: computational biology and experimental validation

doi: 10.1186/s12967-022-03526-0

Figure Lengend Snippet: Effect of RGS1 knockdown on cervical cancer cells. A qRT-PCR was used to detect the expression of RGS1 after knockdown of RGS1 in HeLa and SiHa cells. B Western Blot was used to detect the expression of RGS1 after knockdown of RGS1 in HeLa and SiHa cells. C CCK8 detected the effect of RGS1 on cell proliferation. D Transwell examined the effects of RGS1 on cell migration and invasion. E FCM was used to detect the effect of RGS1 on apoptosis. F The localization of RGS1 in cells was determined by immunofluorescence

Article Snippet: HeLa (CL-0101, Procell, China) and SiHa (CL-0210, Procell, China) were cultured in RPMI 1640 (SH30255.01, HyClone, USA) with 10% fetal bovine serum (FBS) (164,210, Procell, China) at 37 °C and 5% CO 2 .

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Western Blot, Migration, Immunofluorescence

Journal: Journal of Oncology

Article Title: Deficiency of kin17 Facilitates Apoptosis of Cervical Cancer Cells by Modulating Caspase 3, PARP, and Bcl-2 Family Proteins

doi: 10.1155/2022/3156968

Figure Lengend Snippet: Establishment and determination of cervical cells line HeLa and SiHa with kin17 knockdown. (a) morphological features and fluorescence-indicated infection of HeLa Mock , HeLa NC , HeLa KD cells, SiHa Mock , SiHa NC, and SiHa KD cells, ×100. Scale bars = 200 μ m. (b) protein levels of kin17 in HeLa cells and SiHa cells transfected with lentivirus were identified by western blotting, and the densitometric quantification was showed. HeLa KD , HeLa cells transfected with recombinant lentiviral vectors carrying the siRNA-targeting KIN17 gene; HeLa NC , HeLa cells were transfected with the control vector; HeLa Mock , HeLa cells without transfection of the vector. SiHa KD , SiHa cells infected with recombinant lentiviral vectors carrying the siRNA-targeting KIN17 gene; SiHa NC , SiHa cells transfected with the controlled vector; SiHa Mock , SiHa cells without transfection of the vector. NC, negative control; KD, knockdown; ∗ P < 0.05, n = 3.

Article Snippet: Human cervical cancer cell lines HeLa (adenocarcinoma cells; cat. No. TCHu187; GeneChem, Shanghai, China) and SiHa (squamous carcinoma cells; cat. No. TCHu113; GeneChem, Shanghai, China) were cultured in Dulbecco's modified eagle medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Tianjin Kangyuan Biotechnology, Tianjin, China) and 1% antibiotics (Hyclone, USA) consisting of 100 μ g/mL penicillin and streptomycin.

Techniques: Knockdown, Fluorescence, Infection, Transfection, Western Blot, Recombinant, Control, Plasmid Preparation, Negative Control

Journal: Journal of Oncology

Article Title: Deficiency of kin17 Facilitates Apoptosis of Cervical Cancer Cells by Modulating Caspase 3, PARP, and Bcl-2 Family Proteins

doi: 10.1155/2022/3156968

Figure Lengend Snippet: Effect of kin17 knockdown on apoptosis of HeLa and SiHa cells. apoptosis rates of HeLa and SiHa cells with kin17 knockdown and their normal controlled cells were analyzed by flow cytometry (a) representative dot plots with APC fluorescence area (APC-A) as the horizontal ordinate and cumulative data shown, ∗ P < 0.05, n = 3. relative ratios of green/red fluorescence indicating mitochondrial membrane potential of HeLa and SiHa cells were analyzed by flow cytometry (b) representative dot plots with fluorescence and quantification. NC, negative control; KD, knockdown; ∗ P < 0.05, n = 3.

Article Snippet: Human cervical cancer cell lines HeLa (adenocarcinoma cells; cat. No. TCHu187; GeneChem, Shanghai, China) and SiHa (squamous carcinoma cells; cat. No. TCHu113; GeneChem, Shanghai, China) were cultured in Dulbecco's modified eagle medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Tianjin Kangyuan Biotechnology, Tianjin, China) and 1% antibiotics (Hyclone, USA) consisting of 100 μ g/mL penicillin and streptomycin.

Techniques: Knockdown, Flow Cytometry, Fluorescence, Membrane, Negative Control

Journal: Journal of Oncology

Article Title: Deficiency of kin17 Facilitates Apoptosis of Cervical Cancer Cells by Modulating Caspase 3, PARP, and Bcl-2 Family Proteins

doi: 10.1155/2022/3156968

Figure Lengend Snippet: Effect of kin17 knockdown on the expression profile of apoptosis-associated proteins in cervical cancer cells. activities of caspase 3 in kin17 KD and kin17 Mock cells was normalized to the kin17 NC cells (a), the average value of which was set as 100%, ∗ P < 0.05, n = 3. total-length PARP, cleaved PARP, Bim, Bcl-xL, bad, phosphorylated bad in HeLa and SiHa cells were detected via western blotting (b). NC, negative control; KD, knockdown; PARP, poly ADP-ribose polymerase; Bim, Bcl-2 interacting mediator of cell death; Bcl-xL, B-cell lymphoma-xL; Bad, Bcl2 associated death promoter.

Article Snippet: Human cervical cancer cell lines HeLa (adenocarcinoma cells; cat. No. TCHu187; GeneChem, Shanghai, China) and SiHa (squamous carcinoma cells; cat. No. TCHu113; GeneChem, Shanghai, China) were cultured in Dulbecco's modified eagle medium (DMEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Tianjin Kangyuan Biotechnology, Tianjin, China) and 1% antibiotics (Hyclone, USA) consisting of 100 μ g/mL penicillin and streptomycin.

Techniques: Knockdown, Expressing, Western Blot, Negative 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