靶向沉默IL-6对乳腺癌细胞侵袭和迁移能力的影响

Effect of IL-6 silencing on invasion and migration of breast cancer cells

目的:观察白细胞介素6(IL-6)与人乳腺癌MCF-7细胞生长、侵袭和迁移的关系及其作用机制。方法:合成IL-6小干扰RNA(si RNA),并转染至MCF-7细胞中。实验分为空白对照组、阴性对照组和IL-6 si RNA组。通过MTT实验观察沉默IL-6基因表达对细胞活力的影响,Transwell小室法检测细胞侵袭和迁移能力的变化,采用q PCR和Western blot法检测转染后IL-6表达水平的变化,同时采用Western blot法观察上皮-间充质转化(EMT)相关蛋白、信号转导及转录激活因子3(STAT3)、磷酸化STAT3(p-STAT3)、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)的蛋白水平。结果:与对照组相比,转染IL-6 si RNA的MCF-7细胞中IL-6的表达量显著下降(P <0. 05)。沉默IL-6表达显著抑制MCF-7细胞的活力(P <0. 05),并降低其侵袭和迁移能力(P <0. 05),显著抑制N-cadherin和vimentin的表达(P <0. 05),细胞中STAT3无明显变化,p-STAT3、MMP-2和MMP-9的蛋白水平显著降低(P <0. 05)。结论:沉默IL-6表达可能通过抑制EMT,并降低STAT3的磷酸化水平进而抑制MMP-2和MMP-9的分泌,从而减弱乳腺癌MCF-7细胞的活力,并降低其侵袭和迁移能力。

AIM:To investigate the relationship between interleukin-6(IL-6)and viability,invasion and migration of human breast cancer MCF-7 cells and its mechanism.METHODS:IL-6 small interfering RNA(siRNA)was synthesized and transfected into the MCF-7 cells.The cells were divided into blank control group,negative control group and IL-6 siRNA group.The viability of the MCF-7 cells was measured by MTT assay,and the invasion and migration abilities were analyzed by Transwell assay.The expression of IL-6 at mRNA and protein levels was examined by qPCR and Western blot.The protein levels of epithelial-mesenchymal transition(EMT)-related proteins,signal transducer and activator of transcription 3(STAT3),p-STAT3,matrix metalloproteinase-2(MMP-2)and MMP-9 were determined by Western blot.RESULTS:Compared with control groups,IL-6 siRNA decreased the expression of IL-6 significantly and reduced the viability of MCF-7 cells(P<0.05).Silencing of IL-6 expression significantly decreased the invasion and migration abilities of MCF-7 cells(P<0.05),inhibited the expression of EMT-related proteins(P<0.05),and reduced the protein levels of p-STAT3,MMP-2 and MMP-9(P<0.05).CONCLUSION:Silencing of IL-6 expression inhibits the viability of MCF-7 cells and reduces the invasion and migration abilities by inhibiting the expression of EMT-related proteins,decreasing the phosphorylation of STAT3 and thus inhibiting the secretion of MMP-2 and MMP-9.