human cell line pc-9  (DSMZ)

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