目的 探究miR-19a-3p靶向CCND1基因介导FrA-1/IL-6/Stat3通路对乳腺癌侵袭转移的作用机制。方法 癌细胞分组转染(空白组、阴性对照组、miR-19a-3p mimic组、siRNA-CCND1组、miR-19a-3p mimic + siRNA-CCND1组)。分别应用qRT-PCR、Western blot、MTT、划痕实验和Transwell侵袭实验检测相关基因mRNA和蛋白表达及细胞生物学行为变化趋势。结果 人乳腺癌组织和细胞中CCND1和Fra-1的表达明显上升,而miR-19a-3p明显下降,差异有统计学意义(P<0.05)。与空白组和阴性对照组相比,另外3组的miR-19a-3p、CCND1、FrA-1表达水平均上调(P<0.05),miR-19a-3p mimic+siRNA-CCND1组中上调更为明显(P<0.05)。各组细胞在24 h时无明显差异。48 h和96 h各组细胞增殖能力均提高。空白组与阴性对照组细胞增殖能力相比无明显差异。另外3组的细胞增殖能力均明显增强,差异有统计学意义(P<0.05),但miR-19a-3p mimic+siRNA-CCND1组中细胞增殖变化趋势较弱,差异有统计学意义(P<0.05)。与空白组和阴性对照组相比,另外3组细胞迁移、侵袭能力均减弱,差异有统计学意义(P<0.05),且miR-19a-3p mimic+siRNA-CCND1组迁移、侵袭能力最弱,差异有统计学意义(P<0.05)。结论 miR-19a-3p在乳腺癌中呈下降趋势,上调miR-19a-3p可有效抑制CCND1基因表达和FrA-1/IL-6/Stat3通路激活,从而降低乳腺癌细胞侵袭转移。
Abstract
Objective To explore the potential mechanism by which regulation of FrA-1/IL-6/Stat3 signaling pathway by miR-19a-3p targeting CCND1 genes affects the invasion and metastasis of breast cancer.Methods Tumor cells were grouped and transfected (blank group, negative control group, miR-19a-3p mimic group, miR-19a-3p inhibitor group, siRNA-CCND1 group, and miR-19a-3p mimic + siRNA-CCND1 group). Quantitative RT-PCR, Western blot, MTT, scratch test and Transwell invasion test were used to detect the expressions of related genes and proteins, as well as the changes of biological behavior of cells.Results The expressions of CCND1 and Fra-1 in human breast cancer tissues and cells increased, while the expression of miR-19a-3p decreased significantly, and there was significant difference between these groups (All P<0.05). Compared with the blank group and negative control group, the expression level of miR-19a-3p in the miR-19a-3p mimic group and siRNA-CCND1 group increased significantly, while CCND1 and Fra-1 decreased significantly, and cell proliferation, migration, invasion and metastasis decreased (All P<0.05). The expression levels of CCND 1 and Fra-1 in the miR-19a-3p inhibitor group increased, but the level of miR-19a-3p decreased, and the ability of proliferation, migration, invasion and metastasis increased significantly (All P<0.05).In addition, the above-mentioned changes were more significant in the miR-19a-3p mimic + siRNA-CCND1 group than in the miR-19a-3p mimic group and siRNA-CCND1 group, and there was significant difference between these groups (All P<0.05).Conclusions Down-regulation of miR-19a-3p is detected in breast cancer and up-regulation of miR-19a-3p can effectively contain the invasion and metastasis of breast cancer cells by inhibiting the expression of CCND1 genes and activating FrA-1/IL-6/Stat3 pathway
关键词
miR-19a-3p /
CCND1 /
FrA-1/IL-6/Stat3信号通路 /
乳腺癌 /
侵袭转移
Key words
miR-19a-3p /
CCND1 /
FrA-1/IL-6/Stat3 signaling pathway /
breast cancer /
invasion and metastasis
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