| 张来鑫,谢大伟,李青松,等.长链非编码前列腺癌相关转录因子 6靶向微小 RNA-139-3p对骨肉瘤细胞增殖和凋亡的影响[J].安徽医药,2021,25(8):1632-1636. |
| 长链非编码前列腺癌相关转录因子 6靶向微小 RNA-139-3p对骨肉瘤细胞增殖和凋亡的影响 |
| Effect of lncRNA PCAT6 on proliferation and apoptosis of osteosarcoma cells by targeting miR-139-3p |
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| DOI：10.3969/j.issn.1009-6469.2021.08.037 |
| 中文关键词: 骨肉瘤 前列腺癌相关转录因子 6 微小 RNA-139-3p 细胞增殖 凋亡 |
| 英文关键词: Osteosarcoma Prostate cancer associated transcript 6 miR-139-3p Cell proliferation Apoptosis |
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| 中文摘要: |
| 目的探讨长链非编码 RNA(LncRNA)前列腺癌相关转录因子 6(PCAT6)对骨肉瘤细胞增殖和凋亡的影响和分子机制。方法实时荧光定量 PCR(RT-qPCR)检测 20例骨肉瘤组织和与其对应的癌旁组织中 PCAT6和微小 RNA-139-3p(miR139-3p)的表达水平。双荧光素酶报告基因实验和 RT-qPCR验证 PCAT6对 miR-139-3p的靶向调控关系。将 PCAT6小干扰 RNA(si-PCAT6)、 miR-139-3p模拟物(miR-139-3p mimics)分别转染骨肉瘤细胞 SAOS2，细胞计数试剂盒(CCK-8)检测 SAOS2细胞增殖活力，流式细胞术检测 SAOS2细胞凋亡，蛋白质印迹法检测 B细胞淋巴瘤 /白血病 -2(Bcl-2)细胞周期素 D1(Cyclin D1)、 Bcl-2相关 X蛋白( Bax)和 P21、的表达水平。将 si-PCAT6和 miR-139-3p抑制物(anti-miR-139-3p)共转、染至 SAOS2细胞，采用上述方法检测细胞增殖、迁移侵袭能力以及凋亡变化。结果与瘤旁组织相比，骨肉瘤组织中 PCAT6的表达水平［(2.76±0.27)比(1.01±0.09)］显著升高， miR-139-3p的表达水平［(0.51±0.05)比( 1.00±0.08)］显著降低( P <0.05)。 miR-139-3p是 PCAT6的靶基因， PCAT6靶向负性调控 miR-139-3p表达。抑制 PCAT6表达或过表达 miR-139-3p均可下调 SAOS2细胞 CyclinD1和 Bcl-2蛋白表达，上调 p21和 Bax蛋白表达，降低细胞活力，促进细胞凋亡( P <0.05)。抑制 miR-139-3p表达可逆转 si-PCAT6对骨肉瘤 SAOS2细胞增殖抑制和凋亡的影响( P <0.05)。结论 lncRNA PCAT6在骨肉瘤组织中表达上调，抑制 miR-139-3p通过靶向 miR-139-3p抑制骨肉瘤细胞增殖，诱导骨肉瘤细胞凋亡。 |
| 英文摘要: |
| Objective To investigate the effects of long-chain non-coding RNA (LncRNA) prostate cancer associated transcript 6(PCAT6) on proliferation and apoptosis of osteosarcoma cells and its molecular mechanisms.Methods Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of PCAT6 and miR-139-3p in 20 osteosarcoma tissues and their corresponding adjacent tissues. The dual luciferase reporter gene assay and RT-qPCR were applied to verify the targeted and regulatory relationship between PCAT6 and miR-139-3p. PCAT6 small interfering RNA (si-PCAT6) and miR-139-3p mimic (miR-139-3p mimics) were transfected into osteosarcoma cell SAOS2, respectively, and cell proliferation activity was detected by cell proliferation kit (CCK-8), apoptosis was detected by flow cytometry, and the expression levels of B cell lymphoma/leukemia-2 (Bcl-2), Cyclin D1, Bcl-2 related X protein (Bax) and P21 were detected by Western blot. The si-PCAT6 and miR-139-3p inhibitors (anti-miR-139-3p) were co-transfected into SAOS2 cells, and cell proliferation, migration-invasive ability and apoptosis were detected by the above methods.Results Compared with paraneoplastic tissues, the expression level of PCAT6 was significantly higher in osteosarcoma tissues [(2.76±0.27) vs. (1.01±0.09)], while the expression level of miR-139-3p was significantly lower [(0.51±0.05) vs. (1.00±0.08)] (P < 0.05). miR-139-3p was a target gene of PCAT6, and PCAT6 targeted and negatively regulated the expression of miR-139-3p. Inhibition of PCAT6 or overexpression of miR139-3p down-regulated the expression of CyclinD1 and Bcl-2 proteins in SAOS2 cells, up-regulated the expression of p21 and Bax proteins, reduced cell viability and promoted apoptosis (P < 0.05). Inhibition of miR-139-3p reversed the effect of si-PCAT6 on proliferation and apoptosis of osteosarcoma SAOS2 cells (P <0.05).Conclusion The expression of lncRNA PCAT6 is up-regulated in osteosarcoma tissues, inhibiting miR-139-3p inhibits the proliferation and induces apoptosis of osteosarcoma cells by targeting miR-139-3p. |
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