human nsclc cell lines (95-d, a549, hcc827, h358, h1299 pc-9  (Procell Inc)

Journal: Cell Death & Disease

Article Title: SPIN1 accelerates tumorigenesis and confers radioresistance in non-small cell lung cancer by orchestrating the FOXO3a/FOXM1 axis

doi: 10.1038/s41419-024-07225-0

Figure Lengend Snippet: A The relative mRNA expression of SPIN1 in lung adenocarcinoma tissues compared with that in normal lung tissues obtained from the Oncomine database. B SPIN1 protein levels in fresh lung cancer tissues and paired normal tissues ( n = 8) were analysed via western blotting. C The expression of SPIN1 in six NSCLC cell lines and normal bronchial epithelioid cells was detected by western blotting. D Representative images of SPIN1 immunohistochemistry in NSCLC tissues and adjacent nontumorous tissues (Left, scale bar: 100 μm; Right, scale bar: 50 μm). E Quantification of SPIN1 protein expression levels in NSCLC tissues and normal adjacent tissues. F IHC staining scores of the IHC images of NSCLC and adjacent nontumorous tissues. G Kaplan-Meier curves of patients with high and low SPIN1 expression in NSCLC ( n = 120, log-rank test, p < 0.05). n.s., no significant difference, * p < 0.05, ** p < 0.01. The cell experiments were conducted more than 3 times independently, and the data are presented as the means ± standard deviations (SDs).

Article Snippet: Human NSCLC cell lines (95-D, A549, HCC827, H358, H1299 and PC-9), as well as normal human bronchial epithelioid cells (Beas-2B), were purchased from Procell Life Science and Technology (Wuhan, China) and cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium.

Techniques: Expressing, Western Blot, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: SPIN1 accelerates tumorigenesis and confers radioresistance in non-small cell lung cancer by orchestrating the FOXO3a/FOXM1 axis

doi: 10.1038/s41419-024-07225-0

Figure Lengend Snippet: A Western blotting assays were conducted to validate the transfection efficiency of SPIN1 siRNA in A549 (left) and HCC827 (right) cells. B CCK-8 assays were used to evaluate the growth ability of A549 and HCC827 cells transfected with SPIN1 siRNAs. C A colony formation assay was performed in SPIN1-depleted NSCLC cells. Scratch wound healing ( D ) and transwell ( E ) assays were conducted to evaluate the migratory and invasive abilities of NSCLC cells upon SPIN1 depletion. F Representative images of xenograft tumours in two groups (scramble groups and shSPIN1 groups). G Growth curves of xenograft tumours derived from A549 cells expressing scramble or SPIN1 shRNA are presented. H Histograms of tumour weights from the above experiments. ** p < 0.01. At least three replicate experiments were performed, and the final results are presented as the means ± SDs.

Article Snippet: Human NSCLC cell lines (95-D, A549, HCC827, H358, H1299 and PC-9), as well as normal human bronchial epithelioid cells (Beas-2B), were purchased from Procell Life Science and Technology (Wuhan, China) and cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium.

Techniques: Western Blot, Transfection, CCK-8 Assay, Colony Assay, Derivative Assay, Expressing, shRNA

Journal: Cell Death & Disease

Article Title: SPIN1 accelerates tumorigenesis and confers radioresistance in non-small cell lung cancer by orchestrating the FOXO3a/FOXM1 axis

doi: 10.1038/s41419-024-07225-0

Figure Lengend Snippet: Representative images ( A ) and quantification analysis ( B ) of flow cytometry data depicting the cell cycle distribution of NSCLC cells (A549 and HCC827) transfected with NC and SPIN1 siRNAs 6 h after IR (6 Gy). C , D Representative images of neutral comet assays performed 4 h after IR exposure of SPIN1-depleted or control cells. Scale bar: 25 μm. E , F Immunofluorescence staining was performed to detect γ-H2AX foci formation in A549 and HCC827 cells transfected with SPIN1 siRNAs or negative control siRNAs. Scale bar: 10 μm. ** p < 0.01. All the experiments were performed three times independently, and the results are presented as the means ± SDs.

Article Snippet: Human NSCLC cell lines (95-D, A549, HCC827, H358, H1299 and PC-9), as well as normal human bronchial epithelioid cells (Beas-2B), were purchased from Procell Life Science and Technology (Wuhan, China) and cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium.

Techniques: Flow Cytometry, Transfection, Control, Immunofluorescence, Staining, Negative Control

Journal: Cell Death & Disease

Article Title: SPIN1 accelerates tumorigenesis and confers radioresistance in non-small cell lung cancer by orchestrating the FOXO3a/FOXM1 axis

doi: 10.1038/s41419-024-07225-0

Figure Lengend Snippet: A Cells transfected with SPIN1 siRNAs and HA-FOXM1 plasmids were harvested, and the expression of relevant molecules was detected via western blotting. B The proliferation and viability of the indicated NSCLC cells were analysed via CCK-8 assays. Scale bar: 100 μm. C The results of the colony formation assay performed with the indicated cells. D A clonogenic survival assay was used to detect the sensitivity of the indicated cells (A549 and HCC827) transfected with SPIN1 siRNAs or/and FOXM1 plasmids. E The results of neutral comet assay performed in the three groups. Scale bar: 25 μm. F The number and number of Rad51 foci detected by immunofluorescence staining. Scale bar: 10 μm. n.s., no significant difference, * p < 0.05, ** p < 0.01. All experiments were performed independently at least three times, and the results are presented as the means ± SDs.

Article Snippet: Human NSCLC cell lines (95-D, A549, HCC827, H358, H1299 and PC-9), as well as normal human bronchial epithelioid cells (Beas-2B), were purchased from Procell Life Science and Technology (Wuhan, China) and cultured in Roswell Park Memorial Institute 1640 (RPMI-1640) medium.

Techniques: Transfection, Expressing, Western Blot, CCK-8 Assay, Colony Assay, Clonogenic Cell Survival Assay, Neutral Comet Assay, Immunofluorescence, Staining

Journal: Cancers

Article Title: Spironolactone, a Classic Potassium-Sparing Diuretic, Reduces Survivin Expression and Chemosensitizes Cancer Cells to Non-DNA-Damaging Anticancer Drugs

doi: 10.3390/cancers11101550

Figure Lengend Snippet: Spironolactone selectively induces cell death and inhibits the growth of cancer cells. A549, PANC-1, PC-9, and PC-9-OR (osimertinib-resistant) ( a ), cancer stem cell lines (A549 CSLC and PANC-1 CSLC) ( b ), and IMR90 normal human fibroblasts ( c ) were treated without (as control) or with the indicated concentrations of spironolactone (SPL) for three days, and the numbers of viable and dead cells (left panels) as well as the percentage of dead cells (right panels) were then assessed. Values in the graphs represent the means ± SD of triplicate samples of a representative experiment repeated with similar results. * p < 0.05.

Article Snippet: The human non-small cell lung cancer (NSCLC) cell lines A549 and PC-9 were obtained from the Riken BioResource Center (Tsukuba, Japan).

Techniques: Control

Journal: Cancers

Article Title: Spironolactone, a Classic Potassium-Sparing Diuretic, Reduces Survivin Expression and Chemosensitizes Cancer Cells to Non-DNA-Damaging Anticancer Drugs

doi: 10.3390/cancers11101550

Figure Lengend Snippet: Spironolactone decreases survivin expression and survivin reductions imitate the chemosensitizing effects of spironolactone. Cells treated with or without 25 µM spironolactone (SPL) for three days were subjected to an immunoblot analysis for survivin expression ( a , b ). YM155, a pharmacological survivin inhibitor, at the concentration of 10 nM and the knockdown of survivin reduced survivin expression in cancer cells (A549) ( c ). The pharmacological or genetic inhibition of survivin sensitized cancer cells (A549) to chemotherapeutic reagents (GEM, gemcitabine, 0.1 µM; OSI, osimertinib, 2 µM) ( d ). * p < 0.05.

Article Snippet: The human non-small cell lung cancer (NSCLC) cell lines A549 and PC-9 were obtained from the Riken BioResource Center (Tsukuba, Japan).

Techniques: Expressing, Western Blot, Concentration Assay, Knockdown, Inhibition

Journal: Cancers

Article Title: Spironolactone, a Classic Potassium-Sparing Diuretic, Reduces Survivin Expression and Chemosensitizes Cancer Cells to Non-DNA-Damaging Anticancer Drugs

doi: 10.3390/cancers11101550

Figure Lengend Snippet: Spironolactone induces similar effects in GS-Y01, a glioma stem cell line, to those in cancer stem cells (CSCs) of Non-small cell lung cancer (NSCLC) and pancreatic cancer. GS-Y01 cells were treated without (as control) or with the indicated concentrations of spironolactone (SPL) for three days, and the numbers of viable and dead cells (left panel) as well as the percentage of dead cells (right panel) were then assessed ( a ). Cells treated with or without spironolactone (SPL) for three days were subjected to an immunoblot analysis for survivin expression ( b ). Glioma stem cells were treated with the indicated chemotherapeutic reagents (OSI, osimertinib) in the absence or presence of spironolactone (SPL) for three days, and the numbers of viable and dead cells (left panel) as well as the percentage of dead cells (right panel) were then assessed ( c ). Values in the graphs represent the means ± SD of triplicate samples of a representative experiment repeated with similar results. * p < 0.05.

Article Snippet: The human non-small cell lung cancer (NSCLC) cell lines A549 and PC-9 were obtained from the Riken BioResource Center (Tsukuba, Japan).

Techniques: Control, Western Blot, Expressing

Journal: Cancers

Article Title: Spironolactone, a Classic Potassium-Sparing Diuretic, Reduces Survivin Expression and Chemosensitizes Cancer Cells to Non-DNA-Damaging Anticancer Drugs

doi: 10.3390/cancers11101550

Figure Lengend Snippet: Spironolactone sensitizes cancer cells to osimertinib in vivo. Mice (five for each group) were subcutaneously implanted with A549 cells. After the confirmation of tumor formation, mice were treated with or without osimertinib and/or spironolactone as detailed in the Materials and Methods. Tumor volume ( a ) and mouse body weight ( b ) were measured, and the results obtained are presented in the graphs as the means ± SD of each group. * p < 0.05, compared with control group in ( a ) and comparison between spironolactone treatment group and combination group in ( b ).

Article Snippet: The human non-small cell lung cancer (NSCLC) cell lines A549 and PC-9 were obtained from the Riken BioResource Center (Tsukuba, Japan).

Techniques: In Vivo, Control, Comparison