The expression and mechanisms of RNA-9 and RNA-210 in human ovarian cancer cell lines SKOV-3 and SKOV-3/DDP
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摘要: 目的 检验miR-9和miR-210在卵巢癌SKOV-3及SKOV-3/DDP细胞株中的差异表达,观察其对卵巢癌细胞生物学功能的影响并探讨其作用机制,为卵巢癌的靶向治疗积累数据,提供参考方向。 方法 培养人正常卵巢上皮细胞株,卵巢癌SKOV-3和SKOV-3/DDP细胞株,RT-PCR(实时荧光定量聚合酶链反应)检验miR-9和miR-210在各细胞株中的表达差异;将miR-9mimics和miR-210mimics转染至卵巢癌SKOV-3及SKOV-3/DDP中,用CCK-8法检测细胞增殖,Annexin V/PI法测细胞凋亡,细胞划痕实验测细胞迁移能力的改变及细胞侵袭实验测细胞侵袭情况。 结果 miR-9在SKOV3及其耐药株SKOV-3/DDP中低表达,而miR-210在SKOV3及其耐药株SKOV-3/DDP中高表达;miR-9/miR-210过表达抑制3种细胞的增殖;miR-9/miR-210过表达促进细胞发生凋亡;miR-9/miR-210过表达导致3种细胞迁移和侵袭能力下降。 结论 miR-9/miR-210在卵巢癌细胞中起到抑癌基因的作用,过表达miR-9/miR-210能显著抑制卵巢癌细胞的增殖、迁移和侵袭能力并促进肿瘤细胞的凋亡;miR-9/miR-210可以作为卵巢癌及复发性卵巢癌新的潜在治疗靶点。Abstract: Objective To detect the expression of RNA-9 and RNA-210 in human normal ovarian cell line IOSE80, cancer cell line SKOV-3 and its cisplatin-resistant cell line SKOV-3/DDP; To investigate the effects of RNA-9 and RNA-210 on cell multiplication, apoptosis, migration and invasion on SKOV-3 and SKOV-3/DDP; Discuss probable functional mechanisms of RNA-9 and RNA-210 in serous ovarian cancer and provide a way for ovarian cancer targeted therapy finally. Methods Using quantitative real-time PCR to assess the expression of RNA-9 and RNA-210 in normal ovarian cell line, ovarian cell line SKOV-3 and SKOV-3/DDP; Transfect RNA-9 mimics and RNA-210 mimics and its corresponding mimics negative control (mimics NC) into SKOV3 and SKOV-3/DDP respectively. CCK-8 assay, Annexin V/PI assay, Cell scratch experiment, cell invasion experiment were performed to evaluate the influence of overexpression of RNA-9 and RNA-210 on cell multiplication, apoptosis, migration and invasion. Results Compared with normal ovarian cell, the expression of RNA-9 in SKOV-3 and SKOV-3/DDP was higher, and the expression of RNA-210 was lower; CCK-8 assay indicated that the ability of cell multiplication was decreased, Annexin V/PI assay indicated that the ability of cell apoptosis was promoted. Cell scratch experiment showed that the ability of cell migration were significantly decreased, the same with the ability of cell invasion, after Overexpressed either RNA-9 or RNA-210. Conclusion Inclusions miR-9 and miR-210 plays a role of tumor suppressor gene in ovarian cancer cells; The ability of cell apoptosis can be increased by overexpressing the miR-9 or miR-210, and the cell multiplication, migration and invasion can be inhibited not only in ovarian cancer cell but also its cisplatin-resistant cell; miR-9 and miR-210 could be seen as new new potential therapeutic targets for ovarian cancer and recurrent ovarian cancer.
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Key words:
- MiR-9 /
- MiR-210 /
- Ovarian cancer /
- Recurrent ovarian cancer /
- Cell proliferation /
- Invasion /
- Apoptosis /
- miRNA
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