过表达α 2,3-唾液酸转移酶基因对乳腺癌MDA-MB-231细胞侵袭能力的影响

Effects of overexpression of α 2,3-sialyltransferase I on invasion of breast cancer MDA-MB-231 cells

目的研究过表达α2,3-唾液酸转移酶(ST3GalⅢ)基因对乳腺癌MDA-MB-231细胞侵袭能力的影响。方法构建ST3GalⅢ慢病毒表达载体,转染MDA-MB-231细胞。将转染细胞分为空白组(未转染,Mock cells,M)、对照组(转染对照慢病毒,Parent cells,P)和实验组(转染慢病毒载体,ST3GalⅢ,ST3)。用实时定量聚合酶链式反应(RT-PCR)和免疫印迹法检测转染后ST3GalⅢmRNA及蛋白表达,流式细胞术分析转染后ST3GalⅢ催化形成下游产物唾液酸化Lewis抗原(SLe X)变化;用细胞黏附实验、Transwell小室检测MDA-MB-231细胞侵袭能力的影响。结果 RT-PCR与Western bolt显示对照组和实验组中ST3GalⅢmRNA相对表达量分别为0.91±0.04,1.85±0.08;蛋白相对表达量分别为0.94±0.04,1.96±0.10;实验组mRNA和蛋白表达分别为对照组1.84倍和1.76倍(P<0.05);实验组细胞表面SLe X含量为92.86±4.41,为对照组的1.61倍,含量明显升高(P<0.05)。细胞黏附检测结果表明,空白组、对照组和实验组的OD570nm分别为0.32±0.02,0.29±0.01和0.46±0.03;细胞侵袭能力结果表明,空白组、对照组和实验组的OD570nm分别为0.93±0.02,0.81±0.01和1.46±0.13,实验组与对照组比较,差异有统计学意义(P<0.05)。结论过表达ST3GalⅢ基因可提高MDA-MB-231细胞侵袭能力。

Objective To investigate the effect of overexpression of α2,3- sialyhransferase （ST3Gal 11I ） on abilities of invasion in breast cancer cells MDA - MB - 231. Methods ST3Gal ]lI gene lentiviral expression vector was transfected into MDA - MB - 231 cells. Transfected cells were divided into 3 groups： blank group （Mock cells, M） , control group （Parent ceils P） , experimental group （ ST3Gal HI srl＇3 ）. The expression of ST3Gal m mRNA and protein were detected by Quantitative Real - time PCR（ RT - PCR） and Western blot. The content of α2,3- -sialic acids was analyzed by flow cytometry. The ability of migration and invasion in MDA - MB -231 cells were detected by cell adhesion assay and Transwell assay, respectively. Results The expression levels of ST3Gal m mRNA and protein examined by RT - PCR and Western blot in control group and experimental group were 0. 91 ± 0. 04 and 1.85 ± 0. 08 respectively. The expression levels inexperimental group were 1.84 - tbld and 1.76 - fold higher than control group （ P 〈 0. 05 ）. The amount of SLeX on the cell surface in experimental group was 92.86 ± 4.41 and was 1.61 - fold higher than control group, which were significantly increased compared with that in the blank group and control group （P 〈0. 05 ）. OD570nm values of the ad- hesion assay in the in the blank group, the control group and the experimental group were 0. 32 ± 0. 02,0. 29 ± 0.01 and 0. 46±0. 03, respectively, while those of tile invasion assay were 0, 93 ±0. 02,0. 81 ±0. 01 and 1.46 ±0. 13, and there was significant difference between the experimental group and the control group（P 〈0.05）. Conclusion Overexpression of ST3Gal 111 gene could enhance the invasion ability of MDA- MB-231 cells.