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目的探讨左旋棉酚[(-)-gossypol]下调Namalwa细胞中B淋巴细胞刺激因子(B lymphocyte stimulator,BLyS)的可能机制。方法采用CCK8、台盼蓝活细胞染色检测左旋棉酚在体外对Namalwa细胞的生长抑制作用,吖啶橙(acridine orange,AO)染色观察经左旋棉酚处理后细胞自噬的形态特征,Western blot检测细胞中BLyS和自噬蛋白微管蛋白轻链3(LC3)的表达情况。结果左旋棉酚对Namalwa细胞有明显的增殖抑制作用,并且具有时间和剂量依赖性;AO染色后细胞内可观察到明显的细胞自噬小体;Western blot结果显示,左旋棉酚可下调Namalwa细胞中BLyS的表达,并上调LC3Ⅱ蛋白的表达;自噬特异性抑制剂3-methyladenine(3-MA)可显著抑制左旋棉酚对BLyS的下调与LC3Ⅱ的上调作用。结论左旋棉酚可能通过诱导细胞自噬来下调B淋巴细胞中BLyS的表达,从而抑制细胞生长。
Objective To investigate the possible mechanism of L-gossypol [(-) - gossypol] in down-regulating B lymphocyte stimulator (BLyS) in Namalwa cells. Methods The inhibitory effects of L - gossypol on the growth of Namalwa cells in vitro were detected by CCK8 and trypan blue staining. The morphology of autophagy cells was observed by acridine orange (AO) staining. Western blot The expression of BLyS and autophagy protein tubulin light chain 3 (LC3) in cells was detected. Results L-gossypol had a significant inhibitory effect on Namalwa cells in a time-and dose-dependent manner. AO staining showed that significant autophagic cells were observed in the cells. Western blot showed that L-Gossypol could down-regulate Namalwa cells In the BLyS expression and up-regulated the expression of LC3II protein. 3-methyladenine (3-MA), a specific inhibitor of autophagy, significantly inhibited the down-regulation of BLyS and the upregulation of LC3II by L-gp. Conclusion L-gossypol can down-regulate the expression of BLyS in B lymphocytes by inducing autophagy, thereby inhibiting cell growth.