论文部分内容阅读
目的探讨用小干扰核酸(Small interfering RNA,siRNA)靶向沉默神经鞘氨酸激酶1(sphingosine kinase1,SPHK1)基因敲除后对胃癌SGC-7901细胞凋亡的影响。方法人工化学合成SPHK1 siRNA和对照siRNA,分别转染胃癌SGC-7901细胞。Western blot法检测SPHK1、Bcl-2和Bax蛋白的表达;流式细胞术(Flow Cytometry,FCM)检测细胞凋亡的变化。结果 SPHK1 siRNA转染胃癌SGC-7901细胞后,SPHK1蛋白表达明显下调;与对照组相比,SPHK1 siRNA转染组Bcl-2蛋白表达下调了55%(P<0.01),Bax蛋白表达差异无统计学意义,Bcl-2/Bax比值下调54%(P<0.05);SPHK1 siRNA转染组早期凋亡细胞明显增多(P<0.01),晚期凋亡细胞无明显变化。结论 SPHK1特异性siRNA可阻断SGC-7901细胞SPHK1蛋白的表达,并通过影响Bcl-2通路诱导细胞凋亡。
Objective To investigate the effect of silencing sphingosine kinase 1 (SPHK1) knockdown on gastric cancer SGC-7901 cells by using small interfering RNA (siRNA). Methods SPHK1 siRNA and control siRNA were synthesized by manual chemical method and transfected into SGC-7901 cells. Western blot was used to detect the expression of SPHK1, Bcl-2 and Bax protein. Flow Cytometry (FCM) was used to detect the changes of apoptosis. Results SPHK1 siRNA significantly decreased the expression of SPHK1 protein in SGC-7901 cells. Compared with the control group, the expression of Bcl-2 protein in SPHK1 siRNA group was down by 55% (P <0.01), but there was no statistical difference in Bax protein expression The Bcl-2 / Bax ratio was down-regulated by 54% (P <0.05). The number of apoptotic cells in SPHK1 siRNA transfection group was significantly increased at the early stage (P <0.01). Conclusion SPHK1-specific siRNA can block the expression of SPHK1 protein in SGC-7901 cells and induce apoptosis by affecting the Bcl-2 pathway.