【摘要】 目的 分析B细胞活化因子(BAFF)靶向调控细胞凋亡抑制因子(Survivin)对颌下腺细胞(HSG)自噬的影响。方法采用逆转录聚合酶链反应(RT-PCR)检测腮腺组织、颌下腺组织中BAFF、Survivin表达量，并对HSG细胞转染，分为NC组、si-NC组、si-BAFF组、pc DNA-NC组、pc DNA-Survivin组、si-NC-BAFF+anti+pc DNA-NC组、si-NC-BAFF+anti+pc DNA-Survivin组，采用流式细胞仪、Western印迹法检测各组细胞凋亡、自噬。结果 与腮腺组织相比，颌下腺组织中BAFF表达量明显升高，Survivin表达量明显降低(P<0.05)。与HSY细胞相比，HSG细胞中BAFF表达量明显升高，Survivin表达量明显降低(P<0.05)。与NC组、si-NC组相比，si-BAFF组BAFF表达明显降低(P<0.05);与si-NC组相比，si-BAFF组细胞凋亡率明显降低(P<0.05)。与NC组、si-NC组相比，siBAFF组B细胞淋巴瘤(Bcl)-2相关X蛋白(Bax)、自噬微管相关蛋白轻链(LC) 3B、Fas、Fas配体(Fas L)、细胞色素(Cyt)-C表达明显降低，Bcl-2表达明显升高(P<0.05)。与NC组、pc DNA-NC组相比，pc DNA-Survivin组Survivin表达明显升高(P<0.05);与pc DNA-NC组相比，pc DNA-Survivin组细胞凋亡率明显降低(P<0.05)。与NC组、pc DNA-NC组相比，pc DNA-Survivin组Bax、LC3B、Fas、Fas L、CytC表达明显降低，Bcl-2表达明显升高(P<0.05)。双荧光素酶报告试验显示，与pc DNA-NC组相比，Survivin可使BAFF荧光素酶活性明显降低(P<0.05)。与si-NC组相比，抑制Survivin表达可使HSG细胞中Bax、LC3B、Fas、Fas L、Cyt-C表达明显降低，Bcl-2及Survivin表达明显升高(P<0.05);与si-NC-BAFF+anti+pc DNA-NC组相比，si-NC-BAFF+anti+pc DNA-Survivin组Bax、LC3B、Fas、Fas L、Cyt-C表达明显升高，Bcl-2及Survivin表达明显降低(P<0.05)。结论 干扰BAFF表达可对Survivin表达进行调控，抑制HSG凋亡、自噬，延缓疾病的发展。更多还原

【Abstract】 Objective To analyze the effect of B-cell activating factor(BAFF) targeted regulation of apoptosis inhibitory factor(Survivin) on autophagy in submandibular gland cells(HSG).Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression levels of BAFF and Survivin in parotid and submandibular gland tissues. HSG cells were transfected and divided into NC group,si-NC group,si-BAFF group,pc DNA-NC group,pc DNA-NC group,si-NC-BAFF + anti + pc DNA-NC group,si-NC-BAFF+anti+pc DNA-Survivin group. Flow cytometry and Western blot were used to detect apoptosis and autophagy in each group.Results Compared with parotid gland tissue,the expression level of BAFF was significantly increased,while the expression level of Survivin was significantly decreased in submandibular gland tissues(P<0.05). Compared with HSY cells,the expression level of BAFF was significantly increased,while the expression level of Survivin was significantly decreased in HSG cells(P<0.05). Compared with the NC group and si-NC group,the expression level of BAFF was significantly decreased in the si-BAFF group(P<0.05); compared with the si-NC group,the apoptosis rate of cells was significantly decreased in the si-BAFF group(P<0.05).Compared with the NC group and si-NC group,the expression levels of B-cell lymphoma(Bcl)-2 associated X protein(Bax),autophagy microtubule-associated protein light chain(LC) 3B,Fas,Fas ligand(Fas L),cytochrome(Cyt)-C were significantly decreased,while the expression level of Bcl-2 was significantly increased in the si-BAFF group(P<0.05). Compared with the NC group and the pc DNA-NC group,the expression level of Survivin was significantly increased in the pc DNA-Survivin group(P<0.05). Compared with the pc DNA-NC group,the apoptosis rate was significantly decreased in the pc DNA-Survivin group(P<0.05).Compared with the NC group and pc DNA-NC group,the expression levels of Bax,LC3B,Fas,Fas L,and Cyt-C were significantly decreased,while the expression level of Bcl-2 was significantly increased in the pc DNA-Survivin group(P<0.05).The dual-luciferase assay showed that compared with the pc DNA-NC group,the luciferase activity of BAFF was significantly reduced by Survivin(P<0.05). Compared with the si-NC group,the expression levels of Bax,LC3B,Fas,Fas L and Cyt-C were significantly decreased,while the expression levels of Survivin and Bcl-2 were significantly increased in HSG cells by inhibiting the expression of Survivin(P<0.05). Compared with the si-NC-BAFF+anti+pc DNA-NC group,the expression levels of Bax,LC3B,Fas,Fas L and Cyt-C were significantly increased,while the expression levels of Bcl-2 and Survivin were significantly decreased in the si-NC-BAFF+anti+pc DNA-Survivin group(P<0.05). Conclusions Interfering with the expression of BAFF could regulate the expression of Survivin,inhibiting the apoptosis and autophagy of HSG,and delaying the development of the disease.更多还原