陈 明,黄 韬,韩先伟,杨彤涛,周 勇.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[J].现代生物医学进展英文版,2019,19(10):1858-1863. |
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 |
Received:December 10, 2018 Revised:December 31, 2018 |
DOI:10.13241/j.cnki.pmb.2019.10.011 |
中文关键词: 骨肉瘤 miR-155 SOCS1 STAT3 |
英文关键词: Osteosarcoma miR-155 SOCS1 STAT3 |
基金项目:国家自然科学基金项目(81272441) |
Author Name | Affiliation | E-mail | CHEN Ming | Department of Orthopedics, Tangdu hospital, Xi'an, Shaanxi, 710032, China | cming_zhao@163.com | HUANG Tao | Department of Orthopedics, Tangdu hospital, Xi'an, Shaanxi, 710032, China | | HAN Xian-wei | Department of Orthopedics, Tangdu hospital, Xi'an, Shaanxi, 710032, China | | YANG Tong-tao | Department of Orthopedics, Tangdu hospital, Xi'an, Shaanxi, 710032, China | | ZHOU Yong | Department of Orthopedics, Tangdu hospital, Xi'an, Shaanxi, 710032, China | |
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中文摘要: |
摘要 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。 |
英文摘要: |
ABSTRACT 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. |
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