Galectin-3表达抑制对BT-549细胞系增殖和凋亡影响观察  

Influences of inhibiting Galectin-3 by RNA interference technique on the proliferation and apoptosis of BT-549 cells

作　　者：魏杨辉[1,2] 张宗明 杨莉涛[1] 黄耀[1] 刘娟 张卫星[1] 吴爱国 

机构地区：[1]深圳市第四人民医院普外科,广东深圳518033 [2]南方医科大学珠江医院普外科,广东广州510515

出　　处：《中华肿瘤防治杂志》2013年第24期1882-1887,共6页Chinese Journal of Cancer Prevention and Treatment

基　　金：2012年广东省医学科研基金(B2012330);2009年深圳市科技计划(200903202)

摘　　要：目的:观察抑制Galectin-3表达与BT-549细胞系增殖和凋亡的关系,探讨利用RNA干扰技术作为三阴性乳腺癌基因治疗方法的可行性。方法:设计并化学合成Galectin-3小干扰片段并顺势转染乳腺癌细胞系BT-549(为Galectin-3siRNA组),以未转染细胞为空白对照,以转染阴性干扰为阴性对照,荧光倒置显微镜和实时定量PCR验证干扰结果,XTT方法观察干扰后24、48、72和96h各组细胞增殖吸光度(A值),流式细胞仪检测各组细胞凋亡情况,划痕实验和Transwell实验比较不同组间细胞迁移和侵袭情况,以上实验均重复3次。结果:空白对照组、阴性对照组和Galectin-3siRNA组Galectin-3mRNA相对含量分别为(0.078±0.003)、(0.069±0.004)和(0.015±0.001),差异有统计学意义,F=486.97,P<0.001。XTT法检测结果显示,同一组不同时间点之间差异有统计学意义,P值均<0.001;同一时间点各组之间差异亦有统计学意义,P值均<0.001。随着时间推移,空白组和阴性对照组细胞继续生长,Galectin-3siRNA组细胞生长抑制,数量逐渐减少,干扰时间和实验分组之间存在交互效应,F=26.43,P<0.001。流式细胞仪检测结果显示,Galectin-3siRNA组、空白对照组和阴性对照组凋亡率分别为(26.83±2.15)%、(2.73±0.18)%和(5.86±0.51)%,差异有统计学意义,F=24 537.92,P<0.001。空白对照组、阴性对照组和Galectin-3siRNA组愈合率分别为(91.47±4.39)%、(82.36±3.68)%和(52.26±1.95)%,差异有统计学意义,F=213.71,P<0.001。转染Galectin-3siRNA的乳腺癌细胞浸润穿过Matrigel膜的细胞数(169.36±23.71)要比空白对照组(480.80±30.80)和阴性对照组(320.70±28.12)的细胞数显著减少,差异有统计学意义,F=115.20,P<0.001。结论:Galectin-3siRNA成功转染入BT-549细胞,并抑制细胞增殖,诱导细胞凋亡,降低其迁移侵袭能力,为三阴性乳腺癌基因治疗提供实验依据。OBJECTIVE：To observe the relationship of inhibiting Galectin-3 with the proliferation and apoptosis of BT-549 cells and explore the RNA interference technique for triple-negative breast cancer gene therapy. METHODS： Ga- lectin-3 small interfering RNA （siRNA） was constructed by chemistry synthesis. Effective siRNA was transfected into breast cancer cell BT-549（as Galectin-3siRNA group）. Non-transfected cells were used as blank control and negative RNA as negative control. Inverted fluorescence microscope and RT-PCR was used to verify the interference result. Cellprolifera tion （absorhance value） was observed by XTT after interference for 24,48,72 and 96 hours. Cell apoptosis was measured with flow cytomter. Wound healing test and transwell test were performed for cell motility and invasion assay. All the e- periments had been repeated for 3 times. RESULTS：The Galectin-3mRNA relative content in blank control group, negativecontrol group and Galectin-3siRNA group were （0. 078±0. 003）, （0. 069±0. 004） and（0. 015±0. 001） respectively and the differences had statistical significance （F= 486.97, P〈0. 001）. XTT test showed that, the differences between time spots in the same group had statistical significance （P〈0. 001）, while the differences between groups in the same time point also was statistically significant （P=0. 001）. As time going,in the blank group and the negative control group cells continued to grow,but in Galectin-3 siRNA group cell growth was inhibited and ceil number reduced gradually. The inter action effect between the interference time and the experiment group was （F=26.43 ,P%O. 001）. By flow cytometry tes- ting,the apoptosis rates of Galectin-3 siRNA group, blank control group and negative control group were 26. 83%±2. 15%,2.73%±0. 18% and 5. 86± 0. 51 respectively, while the differences was statistically significant （F= 24 537.92,P=0. 001）. Healing rate of blank control group,negative control group and Galeetin-3 siRNA were 91.47±4.39,82.36

关 键 词：乳腺肿瘤 半乳凝集素-3 RNA干扰 细胞增殖 细胞凋亡 

分 类 号：R737.9[医药卫生—肿瘤]