Journal: Cancers

Article Title: Pleiotropic Devitalization of Renal Cancer Cells by Non-Invasive Physical Plasma: Characterization of Molecular and Cellular Efficacy

doi: 10.3390/cancers15020481

Figure Lengend Snippet: NIPP treatment reduces cell proliferation. The human renal cancer cell lines 786-O ( A – C ), Caki-1 ( D – F ) and the non-malignant HREpC cells ( G – I ), were treated with non-invasive physical plasma (NIPP) or the carrier gas argon (Ctrl). The number of living cells was determined at the given times using the CASY Cell Counter and Analyzer and were shown as mean ±SD. Differences between NIPP-treated cells and control cells were tested for significance using the paired t -test (* p < 0.05, ** p ≤ 0.01, *** p ≤ 0.001). For a better overview, the values of the NIPP treatment were shown slightly shifted to the right.

Article Snippet: The human RCC cell lines 786-O and Caki-1 (both Cell Lines Service, Eppelheim, Germany) and the normal non-malignant renal cells HREpC (PromoCell, Heidelberg, Germany) were propagated at 37 °C and 5% CO 2 .

Techniques: Clinical Proteomics, Control

Journal: Cancers

Article Title: Pleiotropic Devitalization of Renal Cancer Cells by Non-Invasive Physical Plasma: Characterization of Molecular and Cellular Efficacy

doi: 10.3390/cancers15020481

Figure Lengend Snippet: Treatment with NIPP-treated cell culture medium. The cell lines 786-O ( A – C ), Caki-1 ( D – F ) and non-malignant cells HREpC ( G – I ) were treated with non-invasive physical plasma (NIPP) activated cell culture media. After 24 h of pre-incubation, the cell culture medium was replaced by NIPP or argon-treated medium and the cells were incubated for further 120 h. The number of living cells was determined at the given timepoints by using the CASY Cell Counter and Analyzer and displayed as mean ±SD. Differences between the NIPP-activated media-treated cells and the control cells were tested for significance using the paired t -test (* p < 0.05, ** p ≤ 0.01, *** p ≤ 0.001). For clarity, the values of the NIPP treatment were shown slightly shifted to the right.

Article Snippet: The human RCC cell lines 786-O and Caki-1 (both Cell Lines Service, Eppelheim, Germany) and the normal non-malignant renal cells HREpC (PromoCell, Heidelberg, Germany) were propagated at 37 °C and 5% CO 2 .

Techniques: Cell Culture, Clinical Proteomics, Incubation, Control

Journal: Cancers

Article Title: Pleiotropic Devitalization of Renal Cancer Cells by Non-Invasive Physical Plasma: Characterization of Molecular and Cellular Efficacy

doi: 10.3390/cancers15020481

Figure Lengend Snippet: Apoptosis assays after NIPP treatment. Malignant cell lines 786-O ( A , B ), Caki-1 ( C , D ) and non-malignant HREpC ( E , F ) were treated with non-invasive physical plasma (NIPP) or carrier gas argon and examined for apoptosis using the established apoptosis detection methods TUNEL assay ( A , C , E ) and Caspase 3/7 activation assay ( B , D , F ). Means ±SD of the relative signal intensity are shown normalized to the respective argon control. The data were tested for significant differences using a paired t -test (* p < 0.05, ** p ≤ 0.01). To validate the TUNEL assay, positive and negative controls were carried out (+ nuclease-pretreated positive control,—unlabeled negative control). Those positive and negative controls were normalized to the mean of the argon controls.

Article Snippet: The human RCC cell lines 786-O and Caki-1 (both Cell Lines Service, Eppelheim, Germany) and the normal non-malignant renal cells HREpC (PromoCell, Heidelberg, Germany) were propagated at 37 °C and 5% CO 2 .

Techniques: Clinical Proteomics, TUNEL Assay, Activation Assay, Control, Positive Control, Negative Control

Journal: Cancers

Article Title: Pleiotropic Devitalization of Renal Cancer Cells by Non-Invasive Physical Plasma: Characterization of Molecular and Cellular Efficacy

doi: 10.3390/cancers15020481

Figure Lengend Snippet: Motility assay and invasion assay after NIPP exposure. Cell lines 786-O ( A , C ) and Caki-1 ( B , D ) were treated with non-invasive physical plasma (NIPP) or the carrier gas argon (Ctrl). Scratch assay ( A , B ): A confluent cell layer was scratched and incubated in a live imagining microscope (Axio Observer Z1, Zeiss, Jena, Germany). Cells were cultured with low-serum conditions. The cell free area was photographed every half hour and measured at the indicated times. The relative area to the starting area at time t = 0 was evaluated. Migration assay ( C , D ): Treated cells were seeded in chambers with a semi-permeable membrane. An FCS gradient was established over the membrane. After 24 h of incubation, the cells on the top and bottom of the membrane were counted, and the invaded cells were compared to the total number of cells. Means ±SD are shown. Differences between the NIPP-treated cells and the control cells were tested for significance by students t -test (* p < 0.05, ** p ≤ 0.01).

Article Snippet: The human RCC cell lines 786-O and Caki-1 (both Cell Lines Service, Eppelheim, Germany) and the normal non-malignant renal cells HREpC (PromoCell, Heidelberg, Germany) were propagated at 37 °C and 5% CO 2 .

Techniques: Motility Assay, Invasion Assay, Clinical Proteomics, Wound Healing Assay, Incubation, Microscopy, Cell Culture, Migration, Membrane, Control

Journal: Cancers

Article Title: Pleiotropic Devitalization of Renal Cancer Cells by Non-Invasive Physical Plasma: Characterization of Molecular and Cellular Efficacy

doi: 10.3390/cancers15020481

Figure Lengend Snippet: NIPP treatment induces loss of dye and ATP leak from cells Cell lines 786-O ( A , D ), Caki-1 ( B , E ) and non-malignant HREpC ( C , F ) were treated with non-invasive physical plasma (NIPP) or the carrier gas argon as a control. Loss of dye assay ( A – C ): Cells were stained with fluorescein diacetate and ethidium bromide. After 15 min of incubation, the cells were analyzed using a flow cytometer. The mean fluorescence intensity (MFI) of the living cells was evaluated. Normalized means + SD were shown. ATP release assay ( E , F ): After treatment cells were sedimented and extracellular ATP concentration in the cell-free supernatant was determined. Relative ATP concentrations + SD were shown. Differences between the NIPP-treated cells and the control cells were tested for significance with the paired t -test (* p < 0.05, ** p ≤ 0.01, *** p ≤ 0.001).

Article Snippet: The human RCC cell lines 786-O and Caki-1 (both Cell Lines Service, Eppelheim, Germany) and the normal non-malignant renal cells HREpC (PromoCell, Heidelberg, Germany) were propagated at 37 °C and 5% CO 2 .

Techniques: Clinical Proteomics, Control, Staining, Incubation, Flow Cytometry, Fluorescence, Release Assay, Concentration Assay

Journal: Oncology Reports

Article Title: GBP2 promotes clear cell renal cell carcinoma progression through immune infiltration and regulation of PD‑L1 expression via STAT1 signaling

doi: 10.3892/or.2023.8486

Figure Lengend Snippet: GBP2 regulates PD-L1 expression at mRNA and protein levels. (A) GBP2 expression in HK-2, Caki-1, Caki-2, 786-O, and ACHN was detected using western blotting. (B) PD-L1 and GBP2 mRNA expression were detected by RT-qPCR with shNS and shGBP2#1 in Caki-1 and 786-O cell lines. (C) PD-L1 and GBP2 mRNA expression were detected by RT-qPCR with vector and GBP2 OE in Caki-1 and 786-O. (D) PD-L1 and GBP2 protein expression were detected by western blotting with shNS and shGBP2#1 in Caki-1 and 786-O cell lines. (E) PD-L1 and GBP2 protein levels were detected by western blotting with vector and GBP2 OE in Caki-1 and 786-O cell lines. Data are expressed as the mean ± SD of three independent experiments. *P<0.05, **P<0.01 and ***P<0.001. GBP 2, guanylate-binding protein 2; PD-L1, programmed death-ligand 1; RT-qPCR, reverse transcription-quantitative PCR; OE, overexpression; sh-, short hairpin.

Article Snippet: A human proximal tubular epithelial cell line (HK2), along with human Caki-2, and Caki-1 cell lines were purchased from iCell Bioscience, Inc.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Plasmid Preparation, Binding Assay, Real-time Polymerase Chain Reaction, Over Expression