SnoN基因在三阴性乳腺癌中的表达及对其体外增殖能力的影响

Expression of SnoN gene in three-negative breast cancer and its effects on proliferation of breast cancer cell line in vitro

目的研究三阴性乳腺癌灶与癌旁组织中SnoN的表达变化,并利用小干扰RNA技术沉默SnoN基因的表达,观察SnoN低表达对于三阴性乳腺癌细胞MDA-MB-231体外增殖及凋亡的变化。方法通过免疫组化实验分别测定三阴性乳腺癌灶及癌旁组织中SnoN的表达变化;将MDA-MB-231细胞分为SnoN siRNA组、controlsiRNA组、None组,脂质体法将SnoN siRNA、control siRNA分别转染入对应组中,None组只加入等量的转染试剂,采用Western blot检测3组MDA-MB-231乳腺癌细胞SnoN蛋白表达的变化;CCK-8检测3组细胞增殖能力的改变,说明其与三阴性乳腺癌之间的关系。结果免疫组化结果显示,三阴性乳腺癌灶中SnoN的表达明显高于癌旁组织,采用SiRNA沉默SnoN基因后,Western blot检测结果显示,SnoN siRNA组的蛋白表达水平与control siRNA组、None组比较显著下降(P<0.01);CCK-8检测结果表明SnoN siRNA组在转染后48、72、96 h后MDA-MB-231乳腺癌细胞增殖能力显著低于相同时间点的2个对照组(P<0.01)。结论 SnoN在与三阴性乳腺癌组织中高表达,沉默SnoN基因能抑制MDAMB-231乳腺癌细胞的增殖,推测SnoN可能影响三阴性乳腺癌的发生和进展,也为临床上对三阴性乳腺癌的诊治提供新的靶点及思路。

Objective To investigate the expression of SnoN in three-negative breast cancer and adjacent tissues,and to observe the effects of low expression of SnoN on proliferation and apoptosis of three-negative breast cancer cell line-MDAMB-231 in vitro. Methods The expression levels of SnoN were detected by immunohistochemistry in three-negative breast cancer and adjacent tissues. The MDA-MB-231 cells were divided into SnoN siRNA group,control siRNA group,none transfection group. The lipofectamine was used to transfect SnoN siRNA and control siRNA into MDA-MB-231 cells,however,the cells in none transfection group were treated only by equivalent transfection reagent,then the expression levels of SnoN protein were detected by Western Blot in the three groups. Moreover the CCK-8 assay was used to detect the changes of proliferation ability of MDA-MB-231 cells,moreover,the correlation between the changes and three-negative breast cancer was analyzed. Results Immunohistochemistry results showed that the expression levels of SnoN in three-negative breast cancer tissues were significantly higher than those in adjacent tissues（ P 0. 01）. After SnoN gene was silenced by siRNA,the expression levels of SnoN in SnoN siRNA group were significantly decreased,as compared with those in control siRNA group and none transfection group（ P 0. 01）. Moreover the proliferation abilities of MDA-MB-231 cells in SnoN siRNA group were significantly decreased at 48 h,72 h,96 h after transfection,as compared with those in control siRNA group and none transfection group（ P 0. 01）. Conclusion The SnoN is highly expressed in three-negative breast cancer,and silencing SnoN gene can inhibit the proliferation of MDA-MB-231 cells,which indicates that SnoN may be correlated with the pathogenesis and development of three-negative breast cancer,moreover,which provides an new target for diagnosis and treatment of threenegative breast cancer in clinical practice.