磁性反义探针在不同转染时间点对SK-Br3肿瘤细胞株c-erbB-2蛋白表达及生长的影响  

Effects of Magnetic c-erbB-2 Antisense Probe on Morphology and Proliferation of SK-Br3 Cancer Cells at Different Transfection Time Points

作　　者：刘海燕[1] 温志鹏[1] 文明[1] 贺海容[1] 谭书德[1] 李少林[1] 

机构地区：[1]重庆医科大学附属第一医院放射科,重庆400016

出　　处：《华中科技大学学报（医学版）》2013年第1期46-50,共5页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目(No.30940021);重庆市自然科学基金资助项目(No.2008BB5209);重庆市卫生局医学科研积极资助项目(No.062025)

摘　　要：目的探讨磁性c-erbB-2反义探针转染SK-Br3细胞株后,在不同时间点对其形态及表达的影响。方法选择磁性c-erbB-2反义探针的含铁终浓度为25μg/mL,分别孵育SK-Br3细胞6、12、24、36及48h。采用普鲁士蓝染色并在光学显微镜及透射电镜下直接观察细胞铁颗粒,用原子吸收光谱法测定细胞内铁含量,锥虫蓝排除实验、CCK8实验观察细胞活性,Western blot检测蛋白质表达,MR成像研究信号强度的变化。结果 SK-Br3细胞在一定范围内(6、12、24h)按时间依赖性摄取磁性c-erbB-2反义探针,当孵育时间为24h时,细胞内铁含量为(18.38±0.28)pg,细胞活力、存活率及细胞内c-erbB-2蛋白表达量明显降低(均P<0.05),MR成像显示细胞T2信号强度明显降低(P<0.05)。36h及48h组各项观察结果与24h组比较,差异均无统计学意义(均P>0.05)。结论磁性c-erbB-2反义探针转染SK-Br3肿瘤细胞24h时,MR成像可以达到最佳显示且明显抑制c-erbB-2癌基因表达。Objective To explore the effects of the magnetic c-erbB-2 antisense probe on morphology and proliferation of SK-Br3 cancer cells at different transfection time points. Methods Breast cancer SK-Br3 cells were transfected with magnetic c- erbB-2 antisense probes containing 25 btg/mL iron and co-cultured for 6,12,24,36 and 48 h, respectively. The distribution and content of iron particles in SK-Br3 cells were determined by Prussian blue staining, electron microscopy and atomic absorption spectrometry respectively. Cell viability was observed by trypan-blue exclusion and CCK-8 assay. The protein expression of cerbB-2 was detected by using Western blot. The signal intensity was measured by MR imaging. Results c-erbB-2 antisense probe was absorbed by SK-Br3 cells in a time-dependent manner within a range of time （6,12,24 h）. When the incubation time was 24 h,the iron content in cells was （18. 38＋0.28） pg,the cell vitality and c-erbB-2 protein expression were reduced significantly （P;0.05） ,and the intensity of T2 signal was weakened （P〈0.05）. The result in the 36 and 48 h groups was not significantly different from that in the 24 h group （P〉0.05）. Conclusion The magnetic c-erbB-2 antisense probe can be effectively transfected into SK-Br3 ceils and markedly inhibit the c-erbB-2 expression 24 h after transfection.

关 键 词：反义寡脱氧核苷酸 超顺磁性氧化铁 分子探针 SK-Br3细胞 

分 类 号：R445.22[医药卫生—影像医学与核医学]