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姜黄素对体内外骨肉瘤细胞增殖和侵袭的影响及其机制研究
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| 篇名： | 姜黄素对体内外骨肉瘤细胞增殖和侵袭的影响及其机制研究 |
| TITLE： | |
| 摘要： | 目的：研究姜黄素对骨肉瘤细胞增殖和侵袭的影响及其机制。方法：体外细胞试验分为对照组（二甲基亚砜），姜黄素低、中、高浓度组（10、20、40 μmol/L），姜黄素高浓度+内源性微小核糖核酸-21模拟物（agomiR-21）组和姜黄素高浓度+agomiR-21阴性对照（NC）组，分别采用CCK-8和Transwell小室法检测各组人骨肉瘤细胞U-2OS和MG-63的增殖[光密度（OD值）]和侵袭能力（穿膜细胞数），分别采用实时荧光定量聚合酶链式反应法和Western blot法检测各组细胞中miR-21及其靶基因PDCD4蛋白的表达。体内实验将MG-63细胞接种至小鼠体内，然后将小鼠分为空白对照组（生理盐水）、姜黄素组（20 mg/kg）、姜黄素+agomiR-21 NC组（20 mg/kg姜黄素溶液+50 mg/kg agomiR-21 NC溶液）、姜黄素+agomiR-21组（20 mg/kg姜黄素溶液+50 mg/kg agomiR-21溶液），每组8只，分别于瘤内注射相应药物，每天1次，连续给药7 d。测量给药后20 d内的肿瘤体积变化，检测第20天瘤组织中miR-21和PDCD4蛋白的表达。结果：与对照组比较，姜黄素低、中、高浓度组U-2OS、MG-63细胞的OD值和穿膜细胞数均明显减小（P＜0.05或P＜0.01）；其中姜黄素中、高浓度组U-2OS、MG-63细胞中miR-21的表达明显减弱（P＜0.05或P＜0.01），PDCD4蛋白表达明显增强（P＜0.01）。与姜黄素高浓度+agomiR-21 NC组比较，姜黄素高浓度+agomiR-21组U-2OS、MG-63细胞的OD值和穿膜细胞数均明显增加（P＜0.01）。与空白对照组比较，姜黄素组和姜黄素+agomiR-21 NC组小鼠给药5 d后肿瘤体积明显减小（P＜0.05或P＜0.01）；瘤组织中miR-21表达明显减弱（P＜0.01），PDCD4蛋白表达明显增强（P＜0.01）。与姜黄素+agomiR-21 NC组比较，姜黄素+agomiR-21组小鼠给药5 d后肿瘤体积明显增加（P＜0.05或P＜0.01）；瘤组织中miR-21表达明显增强（P＜0.01），PDCD4蛋白表达明显减弱（P＜0.01）。结论：姜黄素可抑制人骨肉瘤细胞U-2OS、MG-63的增殖和侵袭，其机制可能与下调miR-21表达有关。 |
| ABSTRACT： | OBJECTIVE： To investigate the effects of curcumin on proliferation and invasion of osteosarcoma cells and its mechanism. METHODS： In in vitro cell assay， osteosarcoma cells were divided into control group（dimethyl sulfoxide）， curcumin low-concentration， medium-concentration and high-concentration groups （10， 20， 40 μmol/L） and curcumin high-concentration+endogenous miR-21 simulacrum （agomiR-21） group， curcumin high-concentration+agomiR-21 negative control （NC） group. The proliferation （OD value） and invasion ability （transmembrane cells number） of human osteosarcoma cells U-2OS and MG-63 were detected by CCK-8 and Transwell chamber method； the expression of miR-21 and its target gene PDCD4 protein were detected by real-time fluorescence quantitative PCR and Western blot assay. In in vivo cell assay， MG-63 cells were inoculated in mice， and the mice were divided into blank control group （normal saline）， curcumin group （20 mg/kg）， curcumin+agomiR-21 NC group （20 mg/kg curcumin solution+50 mg/kg agomiR-21 NC solution）， curcumin+agomiR-21 group （20 mg/kg curcumin solution+50 mg/kg agomiR-21 solution） with 8 mice in each group； they were given intratumoral injection of relevant medicine once a day， for consecutive 7 d. The volume changes of the tumor were measured within 20 d after medication， and the expression of miR-21 and PDCD4 protein in tumor tissue were detected on 20th day. RESULTS： Compared with control group， the OD value and transmembrane cells number of U-2OS and MG-63 cells were decreased significantly in curcumin low-concentration， medium-concentration and high-concentration groups （P＜0.05 or P＜0.01）；the expression of miR-21 was weakened significantly in curcumin medium-concentration and high-concentration groups of U-2OS and MG-63 cells （P＜0.05 or P＜0.01）， while the expression of PDCD4 protein was strengthened significantly （P＜0.01）. Compared with curcumin high-concentration+agomiR-21 NC group， the OD value and transmembrane cells number of U-2OS and MG-63 cells were strengthened significantly in curcumin high-concentration+agomiR-21 group （P＜0.01）. Compared with blank control group， the tumor volume of mice were decreased significantly in curcumin group and curcumin+agomiR-21 NC group （P＜0.05 or P＜0.01）. The expression of miR-21 was decreased significantly in tumor tissue （P＜0.01）， while the protein expression of PDCD4 was strengthened significantly （P＜0.01）. Compared with curcumin+agomiR-21 NC group， the tumor volume of mice were increased significantly in curcumin+agomiR-21 group （P＜0.05 or P＜0.01）. The expression of miR-21 in tumor tissue were strengthened significantly （P＜0.01）， while the expression of PDCD4 protein were weakened significantly （P＜0.01）. CONCLUSIONS： Curcumin can inhibit the proliferation and invasion of human osteosarcoma cells U-2OS and MG-63， the mechanism of which may be associated with down-regulating the expression of miR-21. |
| 期刊： | 2018年第29卷第7期 |
| 作者： | 颜泉 |
| AUTHORS： | YAN Quan |
| 关键字： | 姜黄素；微小核糖核酸-21；骨肉瘤细胞；增殖；侵袭；体内；体外 |
| KEYWORDS： | Curcumin； miR-21； Osteosarcoma cell； Proliferation； Invasion； in vivo； in vitro |
| 阅读数： | 261 次 |
| 本月下载数： | 7 次 |
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