鞘氨醇-1-磷酸信号激活对乳腺癌MCF-7细胞增殖作用机制研究  

Effect and mechanism of sphingosine-1-phosphate signal activation on proliferation of breast cancer MCF-7 cells

作　　者：宋娟[1] 杨晶 王明 张奇[1] 苑家鑫[1] 赵学梅[1] 崔红霞[1] SONG Juan;YANG Jing;WANG Ming;ZHANG Qi;YUAN Jia-xin;ZHAO Xue-mei;CUI Hong-xia(Pharmacy School,Qiqihaer Medical University,Qiqihar 161006,Heilongjiang Province,China)

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006

出　　处：《中国临床药理学杂志》2021年第17期2286-2289,共4页The Chinese Journal of Clinical Pharmacology

基　　金：齐齐哈尔医学科学院基金资助项目(QMSI2017B-07)。

摘　　要：目的研究鞘氨醇-1-磷酸(S1P)信号激活在乳腺癌MCF-7细胞增殖中的作用及其机制。方法(1)将细胞分为空白组和低、中、高3个浓度实验组,实验组用SEW2871(S1P受体激动剂)处理乳腺癌细胞72 h。四甲基偶氮唑蓝法检测细胞增殖。(2)将转染的乳腺癌细胞分为3组:第1转染组、第2转染组和第3转染组。第1转染组(空白质粒转染组)加入慢病毒空白质粒3μL(滴度5.0×10^(12)copy·mL^(-1)),第2转染组(野生型S1P受体过表达组)加入野生型S1P受体基因慢病毒质粒3μL(滴度5.0×10^(12)copy·mL^(-1)),第3转染组(S1P受体磷酸化位点突变过表达组)加入S1P受体磷酸化位点突变基因慢病毒质粒3μL(滴度5.0×10^(12)copy·mL^(-1))。(3)以第1,2和第3转染组细胞作为对照,分别加入W146(S1P受体拮抗剂)、MK2206(Akt信号通路抑制剂)干预。用四甲基偶氮唑蓝法和平板克隆集落形成实验,检测S1P受体过表达、S1P受体阻断及信号通路抑制剂的增殖水平,蛋白质印迹法检测磷酸化信号传导与转录激活因子-3(p-STAT3)蛋白表达。结果(1)空白组和低、中、高3个浓度实验组细胞增殖比率分别为1.00±0.06,1.12±0.14,1.21±0.06和1.07±0.05,中浓度实验组与空白组比较,差异有统计学意义(P<0.05);(2)第1、2和第3转染组细胞增殖比率分别为1.00±0.18,1.29±0.04和1.40±0.15,第2、3转染组与第1转染组比较,差异有统计学意义(P<0.05)。应用W146后,这3组细胞增殖比率分别为1.07±0.06,1.15±0.07和1.11±0.06,第2、3转染组与相应对照比较,差异有统计学意义(P<0.05);第1、2和第3转染组细胞克隆形成数量分别为104.67±13.05,142.67±9.87和131.33±5.86,第2、3转染组与第1转染组比较,差异有统计学意义(P<0.05);应用MK2206后,这3组细胞克隆形成数量分别为(11.33±4.04),(31.00±9.85)和(24.00±7.00)个,第1、2和第3转染组与相应对照比较,差异有统计学意义(P<0.01);(3)应用MK2206后,第1、2和第3转染组的p-STAT3蛋�Objective To study the role of sphingosine 1-phosphate(S1P)signal activation on the proliferation of breast cancer MCF-7cells and its mechanism.Methods(1)Cells were divided into blank group and low,medium and high concentration experimental groups,cells proliferation were detected by MTT assay after treatment with S1P receptor agonist SEW2871 for 72 h.(2)Transfected cells were divided into three groups:transfection-1 group,transfection-2 group,transfection-3 group.The transfection-1 group(blank plasmid group)were treated with 3μL lentiviral blank plasmid(5.0×10^(12)copy·m L^(-1)),transfection-2 group(wild type S1P receptor overexpress group)were treated with 3μL wild type S1P receptor gene lentiviral plasmid(5.0×10^(12)copy·m L^(-1)),transfection-3 group(S1P receptor phosphorylation site mutation overexpress group)were treated with3μL S1P receptor phosphorylation site mutation gene lentiviral plasmid(5.0×10^(12)copy·m L^(-1)).(3)Three groups of transfected cells were used as the control,and W146(S1P receptor antagonist)and MK2206(Akt signaling pathway inhibitor)were added for intervention,respectively.The proliferation of S1P receptor overexpression,S1P receptor blockade and signaling pathway inhibitors were detected by MTT assay and plate clone colony formation assay.The protein expressions of phosphorylation signal transduction and transcriptional activator 3(p-STAT3)were detected by Western blot.Results(1)The growth ratio of MCF-7 cells in blank group and low,medium and high concentration experimental groups were 1.00±0.06,1.12±0.14,1.21±0.06 and 1.07±0.05,comparison between middle concentration experimental group and blank group,the difference was statistically significant(P<0.05).(2)The growth ratio of transfection-1 group,transfection-2 group,transfection-3 group were 1.00±0.18,1.29±0.04,1.40±0.15,respectively,comparison between transfection-2 group,transfection-3 group and transfection-1group,the difference was statistically significant(P<0.05).The growth ratio of MCF-7 cells after treatmen

关 键 词：鞘氨醇-1-磷酸(S1P) 乳腺癌 细胞增殖 S1P受体激动剂 S1P受体拮抗剂 

分 类 号：R97[医药卫生—药品]