sincalide and Multiple-Myeloma

Multiple myeloma (MM) is a malignant proliferative disease of plasma cells, the incidence of which is increasing every year and remains incurable. The enzyme co-activator-associated arginine methyltransferase 1 (CARM1) is highly expressed in a variety of cancers, such as Hodgkin's lymphoma and acute myeloid leukemia, and CARM1 is closely associated with tumor cell proliferation. However, the role of CARM1 in MM has not been elucidated.. In this study, we found that CARM1 is overexpressed in MM and closely associated with poor prognosis in MM. CCK-8 and colony formation assays showed that the proliferation of MM cell lines was downregulated when CARM1 expression was knockdown by specific shRNA. Knockdown of CARM1 reduced the proportion of MM cell lines in the S phase and increased the proportion in G0/G1 phase. RNA-seq analysis of the CARM1-KD cell line revealed that it was closely associated with apoptosis and activated the p53 pathway. CCK-8 and apoptosis results showed that CARM1 knockdown made MM cells more sensitive to standard-of-care drugs.. This study provides an experimental basis for elucidating the pathogenesis of multiple myeloma and searching for potential therapeutic targets.

Topics: Cell Line, Tumor; Cell Proliferation; Humans; Multiple Myeloma; Signal Transduction; Sincalide; Tumor Suppressor Protein p53

To investigate the effects of miR-451a targeting interleukin-6 (IL-6) on the proliferation and apoptosis of multiple myeloma (MM) cells and its potential mechanism via JAK2/STAT3 pathway.. mRNA expression of miR-451a and IL-6R in the plasma of patients with MM and normal controls were determined by RT-qPCR. U266 cells were cultured, transfected with miR-451a mimics, the proliferative ability of U266 cells was determined by CCK-8. Potential targets of miR-451a were predicted with the biological software TargetScan, and the direct relationship between miR-451a and the target IL-6R was analyzed by a dual-luciferase reporter assay. U266 cells were stimulated with IL-6R (100 ng/ml), and the proliferative ability and apoptosis rate were determined by CCK-8 and flow cytometry after 48h.. In the plasma of patients with MM, miR-451a expression was low and IL-6R expression was high. miR-451a targeted and negatively regulated IL-6R. Overexpressing miR-451a inhibited the proliferation and promoted the apoptosis of U266 cells. IL-6R acting on U266 cells promoted the proliferation and inhibited the apoptosis of U266 cells. Overexpressing miR-451a inhibited the activation of JAK2/STAT3 pathway and down-regulating miR-451a promoted the activation of JAK2/STAT3 pathway.. miR-451a targeting IL-6R activates JAK2/STAT3 pathway, thus regulates the proliferation and apoptosis in MM cells.

Topics: Apoptosis; Cell Proliferation; Humans; Janus Kinase 2; MicroRNAs; Multiple Myeloma; Receptors, Interleukin-6; Sincalide; STAT3 Transcription Factor

To study the expression of miR-21 in multiple myeloma (MM) cell lines and plasma cells of patients, and explore the mechanism of miR-21 in MM.. Bone marrow samples from 30 patients with MM and 18 healthy controls were collected. The plasma cells were separated by magnetic beads. MM cell lines (MM1.S cells, RPMI-8226 cells and U266 cells) were cultured. The expression level of miR-21 was detected by real-time fluorescent quantitative PCR (qRT-PCR). After transfection with hsa-miR-21 mimics and hsa-miR-21 inhibitor, the proliferation of MM cells was detected by CCK-8 and cell cloning assay. The target genes regulated by miR-21 were predicted by bioinformatics website. The binding sites of miR-21 and KLF5 were detected by luciferase reporter gene assay. The expression of KLF5 were detected by Western blot and qRT-PCR after hsa-miR-21 mimics and hsa-miR-21 inhibitor were transfected into RPMI-8226 cells. KLF5 plasmid with 3'UTR knockout was synthesized and cotransfected into RPMI-8226 cells with hsa-miR-21 mimics, and the proliferation of MM cells was detected by CCK-8 and cell cloning assay.. Compared with healthy donors, the expression level of miR-21 in plasma cells of patients with MM was significantly increased (P<0.001); the expression of miR-21 in MM cell lines MM1.S, RPMI-8226 and U266 was significantly higher than that in control group (P<0.05). After hsa-miR-21 mimics transfection, the proliferation and the number of colony formation of MM cells was significantly increased, while the proliferation and the number of colony formation of MM cells was decreased after hsa-miR-21 inhibitor transfection (P<0.01). The results of luciferase reporter gene assay showed that miR-21 could bind to 3'UTR of KLF5, and the expression level of KLF5 protein was significantly decreased after hsa-miR-21 mimics transfection. After 3'UTR-knockout KLF5 plasmid and hsa-miR-21 mimics were cotransfected into RPMI-8226 cells, the proliferation of the cells was significantly decreased.. MiR-21 may be involved in regulating the proliferation of MM cells by inhibiting the expression of KLF5.. MiR-21靶向调控KLF5促进多发性骨髓瘤细胞增殖的机制研究.. 探讨多发性骨髓瘤（MM）细胞中miR-21的表达及其作用机制。.. 从30例MM患者及20例健康者骨髓标本中分选出浆细胞，检测miR-21的表达水平；选取MM1.S、RPMI-8226和U266细胞，用qRT-PCR方法检测miR-21的水平；转染hsa-miR-21 mimics和hsa-miR-21 inhibitor后通过CCK-8和细胞克隆实验检测MM细胞增殖情况；预测miR-21调控的靶基因，通过荧光酶素报告基因实验检测miR-21与KLF5的结合位点；利用Western blot和qRT-PCR方法检测转染hsa-miR-21 mimics和hsa-miR-21 inhibitor后KLF5蛋白的表达水平；合成缺失3′UTR的KLF5质粒转染进入过表达miR-21的RPMI-8226细胞，利用CCK-8和细胞克隆实验检测MM细胞增殖情况。.. MM患者浆细胞中miR-21表达水平较正常对照组显著升高（P<0.001）；MM1.S、RPMI-8226和U266细胞中miR-21的表达较对照组也显著升高（P<0.05）。转染hsa-miR-21 mimics后细胞增殖和克隆形成数增加，转染hsa-miR-21 inhibitor后细胞增殖能力明显减弱和克隆形成数明显减少（P<0.01）；荧光酶素报告基因实验结果显示miR-21直接结合KLF5的3′UTR，转染hsa-miR-21 mimics后KLF5蛋白的表达水平明显减低，转染hsa-miR-21 inhibitor后明显升高；将缺失3′UTR的KLF5质粒转染进入miR-21过表达的RPMI-8226细胞后，细胞增殖和克隆形成数显著减弱。.. miR-21可能通过抑制KLF5的表达参与调控MM细胞增殖。.

Topics: 3' Untranslated Regions; Cell Line, Tumor; Cell Proliferation; Humans; Kruppel-Like Transcription Factors; Luciferases; MicroRNAs; Multiple Myeloma; Sincalide