机构地区:[1]南京医科大学第一附属医院乳腺外科,210036
出 处:《中华乳腺病杂志(电子版)》2017年第6期354-360,共7页Chinese Journal of Breast Disease(Electronic Edition)
摘 要:目的探讨淋巴细胞趋化因子(lymphotactin,XCL1)对人乳腺癌MCF-7细胞的表柔比星药物敏感性及侵袭转移能力的影响。方法根据是否进行表柔比星及XCL1干预,将MCF-7细胞分为4组:对照组(MCF-7 NC),表柔比星单药组(MCF-7 E),XCL1单药组(MCF-7 XCL1),联合干预组(MCF-7XCL1+E)。用细胞计数试剂盒8(CCK-8)及平板克隆实验观察XCL1干预后MCF-7细胞对表柔比星药物敏感性的影响。用Transwell法观察XCL1对MCF-7细胞侵袭转移能力的影响,并用免疫荧光法检测对照组和XCL1单药组埃兹蛋白(ezrin)及磷酸化埃兹蛋白(p-ezrin)的表达。用Western blot实验检测4组E钙黏蛋白(E-cadherin)、ezrin及p-ezrin的表达水平。细胞生长抑制率的比较采用析因设计的方差分析,2组均数比较采用t检验,多组均数比较采用单因素方差分析,两两比较用LSD法。结果经XCL1处理2周后,表柔比星对MCF-7细胞的抑制作用降低(组间比较F=23.780,P<0.001;不同浓度比较,F=160.602,P<0.001;浓度因素和分组之间无交互作用,F=1.565,P=0.208)。XCL1刺激MCF-7细胞后,各组(MCF-7 NC、MCF-7 XCL1、MCF-7 E和MCF-7 XCL1+E)细胞克隆形成率分别为(15.63±0.61)%、(19.47±1.94)%、(7.77±0.71)%和(12.37±1.55)%,差异有统计学意义(F=41.925,P<0.001)。迁移实验中MCF-7 XCL1组的细胞穿膜数高于MCF-7 NC组(180.00±20.42比88.00±7.00,t=-7.382,P=0.002),侵袭实验中MCF-7 XCL1组的细胞穿膜数也高于MCF-7 NC组(176.33±8.02比90.33±12.90,t=-9.808,P=0.001)。免疫荧光实验发现XCL1刺激后的MCF-7细胞中p-ezrin蛋白表达明显升高(MCF-7 XCL1:1.08±0.12,MCF-7 NC:0.65±0.11,t=-4.528,P=0.011)。Western blot检测发现,4组比较E-cadherin和p-ezrin表达的差异均有统计学意义(F=6.317、48.517,P均<0.050),ezrin表达差异无统计学意义(F=0.868,P=0.497)。与MCF-7 NC组比较(1.10±0.09),MCF-7 E组、MCF-7XCL1组和MCF-7 XCL1+E组E-cadherin表达均降低(0.65±0.22、0.67±0.14、0.71±0.11,P均<0.050);与MCF-7 NC组比较(0.37±0.07),MCObjective To explore the effect of lymphotactin (XCL1) on epirubicin sensitivity of human breast MCF-7 cells and invasion and metastasis ability of cells. Methods The MCFo7 cells were divided into four groups : the control group ( MCF-7 NC) and three experimental groups ( MCF-7 E group treated with epirubicin, MCF-7 XCL1 group treated with XCL1 and MCF-7 XCLt +E group treated with XCL1 plus epirubicin). CCK-8 assay and plate clone formation assay were used to measure the epirubicin sensitivity of MCF-7 cells after XCL1 treatment. Transwell assay was used to evaluate the migration and invasion ability of MCF-7 cells. The expressions of ezrin and phosphorylated ezrin (p-ezrin) in the control group and MCF-7 XCL1 group were determined by immunofluorescenee assay. The expressions of E-cadherin, ezrin and p-ezrin in four groups were detected by Western blot. The growth inhibition rates of cells were compared using variance analysis of factorial design. The mean values between two groups were compared using t test, the mean values among multiple groups were compared using single factor analysis of variance. Pairwise comparison was performed using LSD method. Results After 2-week treatment of XCL1, the inhibitory effect of epirubicin on MCF-7 cells was decreased (group comparison: F=23. 780,P〈0. 001 ;different concentrations of epirubicin: F= 160. 602, P 〈 0. 001 ; no interaction between grouping and concentration of epirubicin : F = 1. 565, P -- 0. 208 ). The clone formation rates of MCF-7 NC, MCF-7 XCL1, MCF-7 E and MCF-7 XCL1 + E groups were ( 15.63 ±0.61 ) %, ( 19.47 ±1.94 ) %, (7. 77 ± 0. 71 ) % and ( 12. 37 ± 1.55 ) %, respectively, indicating a significant difference (F=41. 925, P〈0. 001 ). The number of cells penetrating the membrane in the MCF-7 XCL1 group was significantly higher than that in the MCF-7 NC group (migration: 180. 00:t:20.42 vs 88. 00± 7.00, t=-7.382, P=0.002; invasion: 176.33±8.02 vs 90. 33±12.90, t =-9808, P=0.001). Im
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