摘要
目的体外实验研究左旋含羞草碱诱导骨肉瘤细胞的凋亡作用,并初步探讨其中的分子机制。方法体外培养骨肉瘤细胞系MG-63及U2-OS,给予0 M,100 M,200 M和400 M的左旋含羞草碱处理,同时在400 M的左旋含羞草碱中加入100 mg/m L柠檬酸铁胺(FAC)处理。CCK-8法检测骨肉瘤细胞系增殖能力,流式细胞术检测细胞凋亡水平,Hoechst染色法检测细胞核固缩比例,蛋白印迹检测左旋含羞草碱及FAC处理后骨肉瘤细胞DNA损伤修复相关蛋白表达。结果 CCK-8法检测细胞增殖,当左旋含羞草碱浓度为400 M时,MG-63及U2-OS的细胞抑制率可分别达到81.1%±1.6%和76.7%±2.1%;流式细胞术检测发现左旋含羞草碱处理48 h后,MG-63及U2-OS的细胞凋亡率分别为54.1%±12.6%和46.2%±14.7%;Hoechst染色法显示,MG-63及U2-OS的细胞核固缩比例分别为53.8%±10.6%和49.2%±8.3%,而加入FAC后,则可以减弱左旋含羞草碱对骨肉瘤细胞的增殖抑制及诱导凋亡的作用;同时,蛋白印迹检测表明DNA损伤修复的关键蛋白表达水平在左旋含羞草碱及FAC处理后均发生显著改变。结论左旋含羞草碱可以有效抑制骨肉瘤细胞增殖并诱导骨肉瘤细胞凋亡,其分子机制与左旋含羞草碱的铁螯合作用有关。
Objective To investigate the mechanism of L-Mimosine induced cell apoptosis in osteosarcoma cell lines. Methods Osteosarcoma cell lines MG-63 and U2-OS were treated with 0 100 200 joM, 400 jiM L-Mimosine,and 400 L-Mimosine is added with lOOmg/mL FAC. CCK-8 was used to test cell proliferation, FITC-Annexin V- PI was carried out to investigate cell apoptosis,Hoechst staining was used to investigate nuclear pyknosis, and western blot was executed to detect the DNA damage and repair related protein expression level. Results CCK-8 results showed that the 400 ／xM L-Mimosine was used to treat MG-63 and U2-OS cell lines, the cell inhibition rates were 81.1%±1.6 and 76.7%±2.1%, respectively. Flow cytometry test showed that, after 48 hours treatment, MG-63 and U2-OS cells apoptosis rates were 54.1%±12.6% and 46.2%±14.7%, nuclearpyknosisrateswere53.8%±10.6%and49.2%±8.3%. Inaddition, FAC can attenuate the effect of L-Mimosine that increasing cell inhibition and apoptosis rate. Western blot showed that the protein expression levels of key regulators involved in DNA damage and repair process were changed significantly after treated with L-Mimosine and FAC. Conclusion L-Mimosine can effectively inhibit the proliferation of osteosarcoma cells via inducing the cell apoptosis process, and the molecular mechanism is related to iron chelation.
作者
江哲珍
祝少博
余黎
汪冰
邓玲珑
魏驰
Jiang Zhezhen, Zhu Shaobo, Yu Li , et al(Zhongnan Hospital of Wuhan University, Wuhan Hubei, 430071, China)
出处
《生物骨科材料与临床研究》
CAS
2018年第2期1-4,共4页
Orthopaedic Biomechanics Materials and Clinical Study
