ail 6  (Bio X Cell)

Journal: International Journal of Oncology

Article Title: Increased interleukin-17A levels promote rituximab resistance by suppressing p53 expression and predict an unfavorable prognosis in patients with diffuse large B cell lymphoma

doi: 10.3892/ijo.2018.4299

Figure Lengend Snippet: Rituximab upregulates the level of IL-17A and the percentages of Th17 cells and IL-17 + Foxp3 + Treg cells in vitro . Co-cultures under 7 different conditions as described in the Materials and methods were defined as groups a-g. (A and B) Representative FACS plots of Th17 cells and IL-17 + Foxp3 + Treg cells in the co-cultures from each group. The numbers displayed are the percentages of Th17 cells and IL-17 + Foxp3 + Treg cells. (C-E) Graphs of the Th17 cells andIL-17 + Foxp3 + Treg cell percentages, and IL-17A in each group. The plots and bars in (B-E) are labeled in the same manner (groups a-g) as in (A); groups a-g in (B-E) represent the same groups as those in (A). Error bars represent standard deviation (SD). Significance was determined using single-factor analysis of variance (one-way ANOVA) with Student-Newman-Keuls/Dunnett's T3 test (3 groups). * P<0.05 and ** P<0.01. The data are from 1 of 3 independent experiments. SU, SU-DHL-4 cells; R, rituximab; aIL-6, neutralizing antibody to IL-6.

Article Snippet: Human recombinant IL-6 (Cat. no. 206-IL) and IL-17A (Cat. no. 7955-IL), and human neutralizing antibodies to IL-6 (aIL-6) (Cat. no. AF-206-NA) and IL-17A (aIL-17A) (Cat. no. AF-317-NA) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vitro, Labeling, Standard Deviation

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Human bone marrow-derived mesenchymal stem cells promote the growth and drug-resistance of diffuse large B-cell lymphoma by secreting IL-6 and elevating IL-17A levels

doi: 10.1186/s13046-019-1081-7

Figure Lengend Snippet: hBMSCs secreted IL-6 into the co-culture supernatants and the IL-6 level was higher in the DLBCL tumor microenvironment. a and b : Levels of IL-6 at different time points in the 1:1 co-culture supernatants of SU-DHL-2/4 cells alone, SU-DHL-2/4 cells with hBMSCs, or hBMSCs alone. Levels of IL-6 ( c and d ), PGE2 ( e and f ), IL-10 ( g and h ), IL-17A ( i and j ), TGF-β ( k and l ), and IL-1β ( m and n ) in the 72 h direct or indirect co-culture supernatants of SU-DHL-2/4 cells with or without hBMSCs at different ratios. o : Graph of IL-6 relative expression (relative IOD) detected by IHC from benign tissues ( n = 48) and tumor tissues ( n = 18). p IHC staining showed various intensities of IL-6 expression in the intercellular spaces of tumor cells or tumor infiltrating lymphocytes (× 400). The data shown represent one of three independent experiments. Error bars represent SD. Significance was determined using two-tailed independent-sample Student’s t/t′ test. (* P < 0.05; ** P < 0.01, compared with the SU-DHL-2/4 group)

Article Snippet: Human recombinant IL-6 and IL-17A, and human neutralizing antibodies to IL-6 (aIL-6) and IL-17A (aIL-17A) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Co-Culture Assay, Expressing, Immunohistochemistry, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Human bone marrow-derived mesenchymal stem cells promote the growth and drug-resistance of diffuse large B-cell lymphoma by secreting IL-6 and elevating IL-17A levels

doi: 10.1186/s13046-019-1081-7

Figure Lengend Snippet: hBMSCs enhanced the growth of DLBCL by secreting IL-6 in vivo. SU-DHL-4 cells were injected subcutaneously into the right flanks of nude mice (BALB/c, n = 24). After the xenograft mouse models were established, hBMSCs (MSCs group, n = 8), IL-6 (IL-6 group, n = 8) or PBS (Control group, n = 8) were injected around the tumors. a : The xenograft mice were killed and the tumor tissues were removed at 28 days after hBMSC, IL-6, or PBS injection. b : Tumor volume, ( c ): relative expression of IL-6 mRNA and ( d ): IL-6 protein expression were measured and detected in each group. e : Representative IHC staining images showing various intensities of IL-6 expression in the intercellular spaces of tumor cells or tumor infiltrating lymphocytes (× 400) in each group. Error bars represent SD. Significance was determined using one-way ANOVA. (* P < 0.05; ** P < 0.01)

Article Snippet: Human recombinant IL-6 and IL-17A, and human neutralizing antibodies to IL-6 (aIL-6) and IL-17A (aIL-17A) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: In Vivo, Injection, Control, Expressing, Immunohistochemistry

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Human bone marrow-derived mesenchymal stem cells promote the growth and drug-resistance of diffuse large B-cell lymphoma by secreting IL-6 and elevating IL-17A levels

doi: 10.1186/s13046-019-1081-7

Figure Lengend Snippet: hBMSCs induced PBMCs differentiation into Th17 and Treg cells, thereby elevating IL-17A and TGF-β levels in the co-culture supernatants. PBMCs from healthy donors were divided into six groups and co-cultured with or without SU-DHL-2/4 cells and/or hBMSCs, and SU-DHL-2/4 cells with hBMSCs were selected as two control groups. All six groups containing PBMCs were co-cultured with IL-2 (100 IU/ml) for 72 h and then detected by FACS. a and c Representative FACS plots of Th17 and Treg cell percentages in PBMCs from six groups. b and d Graphs of Th17 and Treg cell mean percentages from six groups. e , f , and g Graphs of the mRNA levels of related cytokines and transcription factors in the PBMCs from six groups. Levels of IL-17A ( h ), TGF-β ( i ), PGE2 ( j ), IL-6 ( k ), IL-10 ( l ), and IL-1β ( m ) in the co-culture supernatants of eight groups. Cytokines were detected by ELISA. The data shown represent one of three independent experiments. Error bars represent SD. Significance was determined using one-way ANOVA. (* P < 0.05; ** P < 0.01)

Article Snippet: Human recombinant IL-6 and IL-17A, and human neutralizing antibodies to IL-6 (aIL-6) and IL-17A (aIL-17A) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Co-Culture Assay, Cell Culture, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Human bone marrow-derived mesenchymal stem cells promote the growth and drug-resistance of diffuse large B-cell lymphoma by secreting IL-6 and elevating IL-17A levels

doi: 10.1186/s13046-019-1081-7

Figure Lengend Snippet: hBMSCs or IL-6 promoted the growth of DLBCL cells by protecting them from spontaneous or drug-induced apoptosis, and IL-17A reinforced these effects. a Graphs of cell proliferation (OD) in 72-h cultures of SU-DHL-4 cells with or without hBMSCs (1:1), exogenous IL-6 (0.5 ng/ml), IL-17A (0.1 ng/ml), aIL-6 (50 μg/ml) and/or aIL-17A (10 μg/ml). b and c Apoptosis induced by rituximab (10 μg/ml) was protected by IL-6 or IL-17A. DLBCL cell cultures were first treated with exogenous IL-6 (0-5000 pg/ml) or IL-17A (0-5000 pg/ml), and rituximab was added 24 h before apoptosis detection. d Percentages of apoptotic SU-DHL-2 cells in 72-h cultures in the presence of rituximab (10 μg/ml), doxorubicin (2 μM) and Ara-C (2 μM). SU-DHL-2 cells were pre-cultured with exogenous IL-6 (0.5 ng/ml) and IL-17A (0.1 ng/ml) for 48 h before addition of drugs (rituximab, doxorubicin and Ara-C). e and f The sum of apoptotic SU-DHL-4 cell percentages induced spontaneously or by rituximab (10 μg/ml) in all groups. The culture conditions were the same as those in ( a ). g Representative FACS dot plots of apoptotic SU-DHL-4 cell percentages induced spontaneously or by rituximab (10 μg/ml) in all groups. The data shown represent one of three independent experiments. Error bars represent SD. Significance was determined using one-way ANOVA. (* P < 0.05; ** P < 0.01)

Article Snippet: Human recombinant IL-6 and IL-17A, and human neutralizing antibodies to IL-6 (aIL-6) and IL-17A (aIL-17A) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Cell Culture

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Human bone marrow-derived mesenchymal stem cells promote the growth and drug-resistance of diffuse large B-cell lymphoma by secreting IL-6 and elevating IL-17A levels

doi: 10.1186/s13046-019-1081-7

Figure Lengend Snippet: JAK2 and STAT3 phosphorylation were involved in IL-6- or hBMSCs-mediated DLBCL cell growth. Western blot images showing JAK2 ( a ) and STAT3 ( b ) phosphorylation in SU-DHL-2 and SU-DHL-4 cells detected at 0, 15, 30, 60, and 120 min in cultures with IL-6 (0.5 ng/ml) and/or aIL-6 (50 μg/ml). c Western blot images showing JAK2 and STAT3 phosphorylation in SU-DHL-4 cells pretreated without or with hBMSCs (1:1) and/or aIL-6 (50 μg/ml) for 30 min. The data shown represent one of three independent experiments

Article Snippet: Human recombinant IL-6 and IL-17A, and human neutralizing antibodies to IL-6 (aIL-6) and IL-17A (aIL-17A) were obtained from R&D Systems (Minneapolis, MN, USA).

Techniques: Western Blot