Additional file 1: Figure S1. of Interleukin-9 promotes cell survival and drug resistance in diffuse large B-cell lymphoma

IL-9R gene was knockout using lentivirus-mediated RNA interference. The efficiency of IL-9R knockdown was assessed by western blot analysis. The expression of IL-9R protein was obvious decreased in siIL-9R cells than sicontrol cells. (TIF 206 kb

Serum indicators in functional high-risk multiple myeloma patients undertaking proteasome inhibitors therapy: a retrospective study

Multiple myeloma (MM) is a class of malignant plasma cell diseases. An increasing application of autologous stem cell transplantation (ASCT) and anti-myeloma agents represented by proteasome inhibitors (PIs) has improved the response rates and survival of MM patients. Patients progressing within 12 months were recently categorized with functional high-risk (FHR), which could not be clarified by existing genetic risk factors, with poor outcomes. Our study aimed to investigate clinical indices related to FHR and seek prognostic roles in transplant-eligible MM patients. Demographic and individual baseline clinical characteristics were compared by using the Pearson's chi-square and Mann-Whitney U test. Progression-free survival (PFS) and overall survival (OS) were described by Kaplan-Meier estimates and compared using the log-rank test. Logistic regression analysis was used to assess the association of baseline characteristics at MM diagnosis with FHR status. From 18th January 2010 to 1st December 2022, 216 patients were included and divided into two groups according to the FHR status. There was no difference in baseline data between the two groups. Renal impairment (RI, Scr > 2 mg/dL) was common in MM patients and made sense in FHR status. AST levels were validated as independent predictors for FHR status (p = 0.019). Patients with RI or higher AST levels (AST > 40 U/L) tended to have worse outcomes. However, transplants had apparently improved prognoses. Therefore, in the PIs era, transplantations are still effective therapies for transplant-eligible MM patients.</p

Primer Sequences.

<p>Primer Sequences.</p

MTDH and β-catenin are overexpressed and nucleic localization in DLBCL cell lines.

<p>(A) Expression of MTDH (75 kDa) and β-catenin (94 kDa) were detected in the indicated cell lines. PBMCs represent peripheral blood mononuclear cells from healthy samples. Expression of β-actin was used as loading control. (B) Subcellular protein fractionation using the cell lysates of 2 DLBCL cell lines revealed that MTDH and β-catenin were localized in the nucleus (N) and possibly also in the cytoplasm (C) and. The expression of β-actin in the cytoplasm and H3 in the nucleus served as controls for the efficiency of subcellular fractionation.</p

Knockdown of MTDH promotes apoptosis of DLBCL cells.

<p>LY8 cells were transfected with MTDH siRNA or negative control siRNA and cell apoptosis was detected by flow cytometer. Early apoptotic cells were defined as Annexin-V-PE-positive, 7-AAD-negative cells. Columns indicate mean of triplicate determinations; bars, SD. *p<0.05 versus control; ▴p>0.05 versus control.</p

MTDH is overexpressed in DLBCL tumor samples.

<p>(A) Real-time quantitative PCR analysis of MTDH expression in DLBCL (n = 21) and control (n = 25). PCR products were confirmed as a single product at the desired size on agarose gels (1–3: DLBCL; 4–5: Control). Control, reactive hyperplasia of lymph node. β-actin was used as a loading control. (B) Expression of MTDH protein (75 kDa) in DLBCL (1–3) and reactive hyperplasia of lymph node tissues (4–6). Expression levels were normalized with β-actin. (C) Detection of MTDH expression in DLBCL by IHC. (D) Detection of MTDH expression in reactive hyperplasia of lymph node by IHC. Original magnification, ×400. Points indicate mean of triplicate determinations, bars, SD. ****p<0.0001 versus control.</p

MTDH downregulation inhibits the activity of Wnt/β-catenin pathway in DLBCL cells.

<p>(A) Analysis of total β-catenin protein by Western blot analysis in LY8 cells transfected with negative control siRNA and MTDH siRNA. (B) and (C) Expression of cytoplasmic(B) and nuclear (C) β-catenin protein was analyzed in LY8 cells. The expression of β-actin in the cytoplasm and H3 in the nucleus served as controls for the efficiency of subcellular fractionation. Data expressed as mean±SD. *p<0.05 versus control; **p<0.01 versus control; ▴p>0.05 versus control.</p

Upregulation of MTDH enhances proliferation and inhibits apoptosis of DLBCL cells.

<p>(A) LY1 and LY8 cells were either untreated or treated with 250 pg/mL of TNF-α for 48 hours. The expression of MTDH protein was analyzed by Western blot. (B) DLBCL cells were treated with TNF-α at the indicated concentration for 48 hours, and cell proliferation was determined by 3H-TdR incorporation assay. Columns indicate mean of triplicate determinations; bars, SD. (C) LY1(left panel) and LY8(right panel) cells were treated with TNF-α (250 pg/mL for 48 hours) and cell apoptosis was detected by flow cytometer. Early apoptotic cells were defined as Annexin-V-FITC-positive, PI-negative cells. Columns indicate mean of triplicate determinations; bars, SD. *p<0.05 versus control.</p