Journal: Cancer Cell International

Article Title: Exosomal lncRNA SNHG10 derived from colorectal cancer cells suppresses natural killer cell cytotoxicity by upregulating INHBC

doi: 10.1186/s12935-021-02221-2

Figure Lengend Snippet: EMT exosomal lncRNA SNHG10 inhibited NK cell cytotoxicity. A The efficacy of the overexpression of the lncRNA SNHG10 in SW480 cells was verified by qRT-PCR. B The viability of NK92-MI cells was detected by CCK-8 assay. C The cytotoxicity of NK92-MI cells (pretreated with EMT-exo or not) co-cultured with SW480 cells was detected by LDH assay. D The production of IFN-γ from NK92-MI cells was detected by ELISA. The expression of the toxic molecules perforin and granzyme B in NK92-MI cells (pretreated with EMT-exo or not) co-cultured with SW480 cells was measured by qRT-PCR ( E , F ), western blotting ( H ), and immunofluorescence ( H ). GAPDH was used to normalize gene expression. t -test, * P < 0.05, ** P < 0.01

Article Snippet: The human CRC cell line SW480 (RRID: CVCL_0546) (Procell, CL-0223, China) and the human NK cell line NK92-MI (an interleukin [IL]-2-independent NK cell line) (RRID: CVCL_3755) (Procell, CL-0533, China) were obtained from Procell Life Science Technology.

Techniques: Over Expression, Quantitative RT-PCR, CCK-8 Assay, Cell Culture, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Immunofluorescence, Gene Expression

Journal: Cancer Cell International

Article Title: Exosomal lncRNA SNHG10 derived from colorectal cancer cells suppresses natural killer cell cytotoxicity by upregulating INHBC

doi: 10.1186/s12935-021-02221-2

Figure Lengend Snippet: Exosomal lncRNA SNHG10 regulated the function of NK cells through INHBC. A The proliferation of NK92-MI cells was detected by CCK-8 assay. B The cytotoxicity of NK92-MI cells was detected by LDH assay. C The production of IFN-γ from NK92-MI cells was detected by ELISA. D The expression of the toxic molecules perforin and granzyme B in NK92-MI cells was measured by western blotting. GAPDH was used to normalize gene expression. The data were analyzed by ANOVA followed by Tukey’s test, *NC vs. oe-lncSNHG10 exo, # NC vs. si-INHBC. * P < 0.05, ** P < 0.01, *** P < 0.001, ## P < 0.01, ### P < 0.001

Article Snippet: The human CRC cell line SW480 (RRID: CVCL_0546) (Procell, CL-0223, China) and the human NK cell line NK92-MI (an interleukin [IL]-2-independent NK cell line) (RRID: CVCL_3755) (Procell, CL-0533, China) were obtained from Procell Life Science Technology.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Gene Expression

Journal: Cancer Cell International

Article Title: Exosomal lncRNA SNHG10 derived from colorectal cancer cells suppresses natural killer cell cytotoxicity by upregulating INHBC

doi: 10.1186/s12935-021-02221-2

Figure Lengend Snippet: Exosomal lncRNA SNHG10 promoted CRC growth by inhibiting NK cells in vivo. A , B Xenograft tumors of BALB/C mice after injection of LV-lncRNA SNHG10-SW480 and LV-vector-SW480 cells (n = 10 per group). C Tumor volume in the two groups. D Tumor weight of each mouse in the two groups. E , F The levels of the INHBC, perforin, and granzyme B proteins in mice were detected by western blotting. Representative images ( G ) and statistical analysis ( H ) of the flow cytometry of NK1.1 in mice in the two groups. I Representative images of IHC of NK1.1 in mice in the two groups. GAPDH was used to normalize gene expression. t -test, * P < 0.05, ** P < 0.01

Article Snippet: The human CRC cell line SW480 (RRID: CVCL_0546) (Procell, CL-0223, China) and the human NK cell line NK92-MI (an interleukin [IL]-2-independent NK cell line) (RRID: CVCL_3755) (Procell, CL-0533, China) were obtained from Procell Life Science Technology.

Techniques: In Vivo, Injection, Plasmid Preparation, Western Blot, Flow Cytometry, Gene Expression

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Anti-proliferation and apoptosis induction of matrine in NKTCL cells. a NK92 cells and PBMCs were treated with matrine and vindesine at different concentrations for different times. The total viable cells were determined by MTT assay. b NK92 cells were exposed to matrine at different concentrations for 48 h and then determined for apoptotic cells by annexin V and PI staining using flow cytometry. c Percentage (%) of apoptotic cells induced by matrine at various concentrations. Analyses in triplicates. (* p < 0.05, ** p < 0.01 compare to 0 mM group)

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: MTT Assay, Staining, Flow Cytometry

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Matrine induced apoptosis of NKTCL cells via activation of the mitochondrial pathway. a NK92 cells were treated with 1.96 mM matrine for 48 h, followed by western blot. GAPDH was used as a loading control. b The relative intensities of target proteins were normalized to those of GAPDH. Analyses in triplicates. (n.s., not significant; * p < 0.05)

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: Activation Assay, Western Blot, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Matrine inhibited NKTCL cells independent of JAK/STAT3 pathway. a NK92 cells were treated with 1.96 mM matrine for 48 h, followed by western blot for STAT3, p-STAT3 (Tyr705) antibodies. GAPDH was used as a loading control. b The relative intensities of target proteins were normalized to those of GAPDH. Analyses in triplicates. (* p < 0.05, ** p < 0.01)

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: Western Blot, Control

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Decreased c-Myc protein induced by matrine and rescued by proteasome inhibitor. a Effect of matrine on c-Myc protein expression in NK92 cells. NK92 cells were treated with matrine at 1.96 mM for 48 h, and c-Myc protein levels were measured by western blot. b c-Myc mRNA levels in NK92 cells were determined by quantitative RT-PCR at 24 h after matrine treatment at 0, 1.2, 2.4, and 3.6 mM (* p < 0.05, ** p < 0.01 compared to 0 mM group). c CHX chase assay for the half-time of c-Myc. NK92 cells were treated with or without 1.96 mM matrine for 12 h. Cells were then treated with CHX (100 μg/mL) for the indicated minutes, and western blotting was performed. d c-Myc levels were quantified relative to GAPDH levels and graphed as percent c-Myc protein remaining after CHX treatment. Half-lives of c-Myc were calculated from exponential line equations and shown for each treat. e c-Myc protein levels were determined at 6 h post-treatment of MG132 and/or matrine, and ( f ) the relative intensities of target proteins were normalized to those of GAPDH. Analyses in triplicates. (* p < 0.05, ** p < 0.01)

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Matrine inhibited NKTCL cells through CaMKIIγ/c-Myc pathway. a NK92 cells were treated with 1.96 mM matrine for 48 h, followed by western blot for c-Myc, p-c-Myc (Ser62), CaMKIIγ and LMP1 antibodies. GAPDH was used as loading control. b The relative intensities of target proteins were normalized to those of loading control (** p < 0.01). c LMP1, RUNX3, EZH2, miR-26a, miR-26b and miR-101 transcription levels in NK92 cells were determined by quantitative RT-PCR at 24 h after matrine treatment at 0, 1.2, 2.4, and 3.6 mM (* p < 0.05, ** p < 0.01 compared to 0 mM group). Analyses in triplicates

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: Western Blot, Control, Quantitative RT-PCR

Journal: BMC Complementary Medicine and Therapies

Article Title: Matrine inhibits the growth of natural killer/T-cell lymphoma cells by modulating CaMKIIγ-c-Myc signaling pathway

doi: 10.1186/s12906-020-03006-2

Figure Lengend Snippet: Cartoon diagram of the mechanisms for growth suppression of NKTCL cells by matrine. Matrine inhibits the transcription of c-Myc through the downregulated LMP1 protein. Matrine downregulates c-Myc phosphorylation at Ser62 through CaMKIIγ inhibition, and then promotes the destabilization and degradation of c-Myc protein in a proteasome-dependent manner

Article Snippet: The human NKTCL NK92 cell line was obtained from the DSMZ collection (Germany) and maintained in MEM alpha medium supplemented with 12.5% fetal bovine serum (Gibco), 12.5% horse serum (Gibco), 10 ng/mL IL-2 (PeproTech, USA) in a humidified 5% CO 2 atmosphere at 37 °C.

Techniques: Phospho-proteomics, Inhibition