Objective-To investigate cystathionine β synthase (CBS)/hydrogen sulfide (H₂S) signaling in multiple myeloma (MM) patients and to identify its effect on the proliferation of U266 cells.
Methods-Bone marrow samples of 19 MM patients and 23 healthy donors were collected. qRT-PCR was performed to measure the mRNA expression levels of H₂S synthases, cystathionine β synthase, and cystathionine γ lyase. ELISA assays quantified the amount of H₂S produced by the two enzymes CBS and CSE. CCK-8 experiment was used to investigate the influence of the CBS inhibitor amino oxyacetic acid and the CSE inhibitor propargylglycine on the proliferation of U266 cells. Flow cytometry and western blotting were performed to determine the effects of AOAA, PAG, and NaHS on cell cycle distribution as well as Caspase-3 and Bcl-2 expression.
Results-Patients with MM had higher level of CBS compared with healthy donors. AOAA significantly inhibited cell proliferation in both a time and concentration dependent characteristic, whereas PAG does not. After 24 hours of treatment, AOAA significantly elevated the G0/G1 phase proportion of cells, and reduced the cell distribution in both S and G2/M phases, while NaHS accelerated cell cycle progression by reducing the relative number of cells in G0/G1 phase and increasing the proportion of cells in the G2/M phase. Moreover, AOAA abolished the impact of NaHS on cell cycle progression of U266 cells. AOAA treatment also led to a significant decrease in Bcl-2 expression and dramatic increase in Caspase-3 expression, though NaHS reversed these effects.
Conclusion-CBS/H₂S system might have a certain effect on the proliferation and apoptosis of MM cells.

中文翻译：

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多发性骨髓瘤中的胱硫醚β合酶/硫化氢信号调节细胞增殖和凋亡。

目的-研究多发性骨髓瘤（MM）患者的胱硫醚β合酶（CBS）/硫化氢（H ₂ S）信号传导，并确定其对U266细胞增殖的影响。
方法-收集19例MM患者和23名健康供体的骨髓样本。进行qRT-PCR以测量H ₂ S合酶，胱硫醚β合酶和胱硫醚γ裂合酶的mRNA表达水平。ELISA分析定量了H ₂的量S由两种酶CBS和CSE产生。采用CCK-8实验研究了CBS抑制剂氨基氧乙酸和CSE抑制剂炔丙基甘氨酸对U266细胞增殖的影响。进行流式细胞术和蛋白质印迹来确定AOAA，PAG和NaHS对细胞周期分布以及Caspase-3和Bcl-2表达的影响。
结果-MM患者的CBS水平高于健康捐献者。AOAA在时间和浓度依赖性特征上均显着抑制细胞增殖，而PAG则不。处理24小时后，AOAA显着提高了细胞G0 / G1期的比例，并减少了S和G2 / M期的细胞分布，而NaHS通过减少G0 / G1期的相对细胞数来加速细胞周期进程并增加了G2 / M期细胞的比例。此外，AOAA废除了NaHS对U266细胞的细胞周期进程的影响。AOAA处理还导致Bcl-2表达显着下降和Caspase-3表达显着增加，尽管NaHS逆转了这些作用。
结论-CBS / H ₂S系统可能对MM细胞的增殖和凋亡有一定影响。