miR-155靶向SOCS1对骨肉瘤Saos2细胞的增殖、侵袭和迁移能力的影响

miR-155 Regulates Proliferation, Migration and Invasion of Osteosarcoma Saos2 Cells by Targeting SOCS1

目的:探讨microRNA-155(miR-155)对骨肉瘤Saos2细胞增殖、侵袭和迁移的影响以及其作用机制。方法:利用实时荧光定量(qRT-PCR)实验检测miR-155在正常成骨细胞与骨肉瘤Saos2细胞中的表达水平,以及miR-155-mimic、miR-155-inhibitor的转染效率。采用CCK-8实验检测细胞的增殖能力,Transwell实验和划痕实验分别检测Saos2细胞的侵袭和迁移能力,Western blot检测细胞内的STAT3磷酸化水平以及SOCS1表达水平,双荧光素酶报告基因实验进行靶基因验证。结果:miR-155在骨肉瘤Saos2细胞中表达明显高于正常成骨细胞(P<0.001)。在分别转染miR-155-mimic和miR-155-inhibitor后,Saos2细胞内miR-155表达水平明显上调和下降(P<0.001)。过表达miR-155可促进Saos2细胞增殖、侵袭和迁移,降低SOCS1的蛋白水平,上调STAT3的磷酸化水平,差异均具有统计学意义。相反,降低miR-155水平可抑制Saos2细胞的增殖、侵袭和迁移能力,差异均具有统计学意义。结论:骨肉瘤Saos2细胞中高表达的miR-155可以通过抑制SOCS1表达来激活STAT3信号通路进而促进细胞的增殖、侵袭和迁移,因此,靶向抑制miR-155表达可以作为潜在治疗骨肉瘤的途径。

Objective: To investigate the effects and the mechanism of action of microRNA-155(miR-155) on proliferation,invasion and migration of osteosarcoma Saos2 cells. Methods: The expression of miR-155 in normal osteoblasts and osteosarcoma Saos2 cells was detected by real-time PCR, and the transfection efficiency of miR-155-mimic and miR-155-inhibitor was measured by qRT-PCR. CCK-8 assay was used to evaluate the cell proliferation. Transwell assay and wound healing assay were used to detect the invasion and migration ability of Saos2 cells, respectively. SOCS1 and p-STAT3 protein expression was examined by using western blot assay. Dual luciferase reporter gene assay was performed to determine the target gene. Results: The expression of miR-155 was significantly higher in osteosarcoma Saos2 cells than normal osteoblasts(P <0.001). After transfection with miR-155-mimic or miR-155-inhibitor, miR-155 expression of Saos2 cells was significantly upregulated or downregulated, respectively(P <0.001).Overexpression of miR-155 promoted proliferation, invasion and migration of Saos2 cells, and decreased the protein level of SOCS1, as well as increased the phosphorylation level of STAT3. The difference was statistically significant(P<0.05). Conversely, downregulation of miR-155 inhibited the proliferation, invasion, and migration of Saos2 cells. The difference was statistically significant(P <0.05).Conclusion: miR-155, which is highly expressed in osteosarcoma Saos2 cells, can activate the STAT3 signaling pathway by inhibiting the expression of SOCS1, thereby promotes cell proliferation, migration and invasion. Therefore, targeting miR-155 therapy may be a potential approach for osteosarcoma.