摘要
目的探讨Sixl基因在乳腺癌组织表达及RNA干扰其表达对癌细胞增殖侵袭能力的影响。方法通过RT-PCR及Western印迹分别检0n,4Sixl在乳腺癌组织的mRNA及蛋白表达；通过脂质体Lipofectami．neTM2000将Sixl的小干扰RNA（siRNA）（Sixl-siRNA组）转染人乳腺癌MDA-MB．231细胞，设置空白对照组和阴性对照组（NC组），转染48h后检测各组细胞中Sixl的蛋白表达；细胞计数试剂盒（CCK8）法检测细胞活力，Transwell小室检测细胞侵袭能力；Western印迹检测细胞增殖蛋白ki67、侵袭相关蛋白基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）及信号转导与转录因子3（STA33）和磷酸化的STAT3的蛋白表达。结果乳腺癌组织中Sixl的mRNA及蛋白表达均显著高于癌旁组织（P〈0．05）；转染Sixl的siRNA组细胞Sixl的蛋白表达显著低于空白对照组（P〈0．05）；与空白对照组比较，Sixl-siRNA组细胞活力及侵袭能力均显著低于空白对照组，ki67、MMP-2、MMP-9和p-STAT3蛋白表达均显著低于空白对照组（P〈0．05），3组间STAT3的蛋白表达无显著性差异（P〉0．05）。结论乳腺癌组织中Sixl高表达．抑制其表达可通过下调STAT3信号降低细胞的活力及侵袭能力。
Objective To investigate the expression of Six1 gene in breast cancer and the effect of its down-regulated expression on the proliferation and invasion ability. Methods RT-PCR and Western blotting were used to detect mRNA and protein levels of Sixl in breast cancer. Human breast cancer MDA-MB-231 cells were transfected with Sixl siRNA （ Sixl-siRNA group） by using liposome Lipofectamine2000TM, and the blank control group and negative control group （NC group） were set up. The expression of Sixl protein in each group was detected after cells were transfected for 48 h. Cell proliferation was detected by CCK8 assay, the invasion ability wby Transwell and the expression of Ki67, MMP-2, MMP-9, STAT3 and p-STAT3 protein by Western blotting. Results The mRNA and protein expressions of Sixl in breast cancer were significantly higher than in adjacent tissues （P 〈 0. 05 ） ; the protein expression of Sixl in siRNA group transfected with Sixl were significantly lower than in the blank control group （P 〈 0. 05） . Compared with the control group, cell viability and invasion ability in Six/-siRNA group were significantly lower than in the control group, the expression of Ki67, MMP-2, MMP-9 and p-STAT3 protein were significantly lower in Sixl -siRNA group than in the control group （P 〈 0.05 ） , the protein expression of STAT3 showed no significant differences among the three groups （P 〉 0. 05） . Conclusion Sixl is highly expressed in breast cancer tissues, and inhibition of its expression can reduce cell proliferation and invasion by down-regulating STAT3 signaling.
作者
陈露
CHEN Lu(Department of Pathology,Ninth People＇s Hospital of Wuxi,Wuxi,Jiangsu,214000,China)
出处
《医学分子生物学杂志》
CAS
2018年第5期315-319,共5页
Journal of Medical Molecular Biology
关键词
Sixl基因
乳腺癌
增殖
侵袭
信号通路
Sixl gene
breast cancer
proliferation
invasion
signal pathway
