human nk cell lines nk92  (ATCC)

Journal: Cancer Cell International

Article Title: LINC00665 promotes the progression and immune evasion of lung cancer by facilitating the translation of TCF7 protein through dependence on IRES

doi: 10.1186/s12935-024-03411-4

Figure Lengend Snippet: Knockdown of LNC00665 Promotes NK Cell Cytotoxicity Against Lung Cancer Cells. NK92 cells were stimulated with IL-2, and then activated NK92 cells (effector cells) were co-cultured with A549/H1299 cells in each treatment group. ( A ) Detection of lactate dehydrogenase (LDH) levels in A549/H1299 cells using the CytoTox96 non-radioactive cytotoxicity assay (Promega, Madison, WI, USA) to assess NK cell cytotoxicity against lung cancer cells. ELISA was used to measure the secretion of ( B ) IFN-γ and ( C ) TNF-α in the cell culture supernatant of the co-culture system. ( D ) NK cell migration was evaluated in transwell assays allowing for only active migration of NK cells, laid onto the upper insert, towards the bottom where A549/H1299 cells were cultured. ( E-G ) Purified NK cells were co-cultured with A549/H1299 cells for 5–6 days at a 1:1 ratio and evaluated for proliferation (Cell-Trace or Ki67), IFN-γ production, and degranulation (CD107a). NK cell IFN-γ and degranulation were assessed following stimulation with PMA and ionomycin for 6 h prior to staining. The cell experiments were repeated three times, and data are presented as mean ± standard deviation. Multiple group comparisons were performed using one-way ANOVA, and post hoc analysis was conducted using Tukey’s multiple comparisons test. ** indicates P < 0.01, *** indicates P < 0.001

Article Snippet: The human NK cell line NK92 cells (ATCC) were cultured in MEMa supplemented with 12.5% FBS, 2 mM L-glutamine, and 12.5% horse serum (Gibco).

Techniques: Knockdown, Cell Culture, Cytotoxicity Assay, Enzyme-linked Immunosorbent Assay, Co-Culture Assay, Migration, Purification, Staining, Standard Deviation

Journal: Cancer Cell International

Article Title: LINC00665 promotes the progression and immune evasion of lung cancer by facilitating the translation of TCF7 protein through dependence on IRES

doi: 10.1186/s12935-024-03411-4

Figure Lengend Snippet: Partial Reversal of the Enhanced NK Cell Cytotoxicity Against Lung Cancer Cells by LINC00665 Knockdown Through Overexpression of HHLA2. ( A ) Western blot analysis of HHLA2 expression. ( B ) CCK-8 assay measuring cell viability. ( C ) Flow cytometry analysis of cell apoptosis. ( D ) Transwell assay evaluating invasion and migration. ( E-H ) Proliferation (Cell-Trace or Ki67), degranulation (CD107a), and IFN-γ production of NK92 cells co-cultured with A549/H1299 cells. Cell experiments were repeated three times, and data are presented as mean ± standard deviation. The comparison between the two groups was performed using an independent sample t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001

Article Snippet: The human NK cell line NK92 cells (ATCC) were cultured in MEMa supplemented with 12.5% FBS, 2 mM L-glutamine, and 12.5% horse serum (Gibco).

Techniques: Knockdown, Over Expression, Western Blot, Expressing, CCK-8 Assay, Flow Cytometry, Transwell Assay, Migration, Cell Culture, Standard Deviation, Comparison

Journal: Cancer Cell International

Article Title: LINC00665 promotes the progression and immune evasion of lung cancer by facilitating the translation of TCF7 protein through dependence on IRES

doi: 10.1186/s12935-024-03411-4

Figure Lengend Snippet: In Vivo Deletion of LINC00665 Suppresses Tumor Growth and Enhances NK Cell Sensitivity. ( A-C ) Tumor growth in a xenograft model established by subcutaneously injecting LINC00665-knockdown A549 cells into NICD-SOD mice, with further intravenous injection of activated NK92 cells on days 3 and 7 post-implantation. ( D ) Ki-67 staining of tumor sections. ( E ) Flow cytometry analysis of CD107a + NK cells in the tumor. Data are presented as mean ± standard deviation. The comparison between the two groups was performed using an independent sample t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001

Article Snippet: The human NK cell line NK92 cells (ATCC) were cultured in MEMa supplemented with 12.5% FBS, 2 mM L-glutamine, and 12.5% horse serum (Gibco).

Techniques: In Vivo, Knockdown, Injection, Staining, Flow Cytometry, Standard Deviation, Comparison