SPK1通过调节Bcl-2/Bax途径干扰LLC细胞凋亡  

SPK1 interferes with LLC cell apoptosis by regulating Bcl-2/Bax pathway

作　　者：吕冰洁[1] 赵洁[2] 郝东[1] 王晓芝[1] 王涛[1] 李洪波[1] 杨阳[2] LV Bing-jie;ZHAO Jie;HAO Dong;WANG Xiao-zhi;WANG Tao;LI Hong-bo;YANG Yang(Department of Respiratory and Critical Care Medicine,Affiliated Hospital of Binzhou Medical College,Binzhou 256603,China;Department of Geriatrics,Affiliated Hospital of Binzhou Medical College,Binzhou 256603,China)

机构地区：[1]滨州医学院附属医院呼吸与重症医学科,山东滨州256603 [2]滨州医学院附属医院老年医学科,山东滨州256603

出　　处：《中国病理生理杂志》2019年第5期950-954,960,共6页Chinese Journal of Pathophysiology

基　　金：山东省高等学校科技计划项目(No.J17KB080)

摘　　要：目的:研究鞘氨醇激酶1(SPK1)是否通过调节Bcl-2/Bax途径干扰小鼠Lewis肺癌(LLC)细胞的凋亡。方法:通过构建SPK1基因小干扰RNA(siRNA)真核表达载体,将siRNA转染至LLC细胞,在荧光显微镜下观察LLC细胞转染的情况。采用流式细胞术检测转染后LLC细胞的凋亡率,Western blot法检测转染后LLC细胞中SPK1、Bcl-2和Bax蛋白表达水平,ELISA法检测Bax和Bcl-2的蛋白表达。结果:转染后的LLC细胞在荧光显微镜下发出绿色荧光。siRNA-SPK1组细胞凋亡明显高于siRNA-SPK1-Neg组(P<0.01)。Western blot结果显示, siRNA-SPK1组中Bax蛋白表达明显较siRNA-SPK1-Neg组高,Bcl-2蛋白表达较siRNA-SPK1-Neg组低。ELISA结果显示siRNA-SPK1组中Bax表达水平显著高于siRNA-SPK1-Neg组(P<0.01), siRNA-SPK1组中Bcl-2表达水平显著低于siRNA-SPK1-Neg组(P<0.01)。结论:SPK1在LLC细胞中的表达与细胞凋亡率有关。SPK1可能是通过Bcl-2/Bax途径干扰LLC细胞凋亡。AIM:To investigate whether sphingosine kinase 1(SPK1)interferes with apoptosis of Lewis lung cancer(LLC)cells by regulating the Bcl-2/Bax pathway.METHODS:The SPK1 gene siRNA eukaryotic expression vector was constructed,and transfected into the LLC cells.The transfected LLC cells was observed under a fluorescence microscope.The apoptotic rate of LLC cells after transfection was analyzed by flow cytometry.The expression levels of SPK1,Bcl-2 and Bax in LLC cells after transfection were detected by Western blot.The protein levels of Bax and Bcl-2 were measured by ELISA.RESULTS:Transfected LLC cells emitted green fluorescence under a fluorescence microscope.Apoptosis in siRNA-SPK1 group was significantly higher than that in siRNA-SPK1-Neg group(P<0.01).Western blot analysis showed that the expression of Bax in siRNA-SPK1 group was significantly higher than that in siRNA-SPK1-Neg group,and the expression of Bcl-2 was lower than that in siRNA-SPK1-Neg group.The ELISA results showed that the protein level of Bax in siRNA-SPK1 group was significantly higher than that in siRNA-SPK1-Neg group(P<0.01),and the protein level of Bcl-2 in siRNA-SPK1 group was significantly lower than that in siRNA-SPK1-Neg group(P<0.01).CONCLUSION:The expression of SPK1 in LLC cells is related to the apoptotic rate.SPK1 may interfere with the apoptosis of LLC cells via Bcl-2/Bax pathway.

关 键 词：鞘氨醇激酶1 Bcl-2/Bax通路 非小细胞肺癌 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤] R730.23[医药卫生—临床医学]