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长链非编码RNA HOXD-AS1介导骨肉瘤细胞阿霉素耐药的机制研究 被引量:2

Effects of lncRNA HOXD-AS1 on doxorubicin chemotherapy-resistant of osteosarcoma
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摘要 目的:观察长链非编码RNA HOXD-AS1对骨肉瘤(osteosarcoma,OS)细胞增殖、凋亡能力以及对阿霉素化疗耐药的影响,并探讨其机制。方法:首先采用实时荧光聚合酶链反应法(real-time polymerase chain reaction,RT-PCR)检测人正常成骨细胞和OS细胞系以及阿霉素耐药OS细胞系中lncRNA HOXD cluster antisense RNA 1(lncRNA HOXD-AS1)和P-gp(P-glycoprotein)的表达水平;在骨肉瘤细胞中抑制lncRNA HOXD-AS1表达后,通过RT-PCR和Western-blot检测P-gp的表达水平;在骨肉瘤细胞中抑制lncRNA HOXD-AS1表达后,再过表达P-gp,应用CCK-8法检测转染后各组吸光值以评价增殖率,应用Annexin V/PI染色流式细胞术检测各组细胞凋亡率,在培养基中加入阿霉素检测各组细胞存活率。结果:与正常成骨细胞相比,OS细胞中lncRNA HOXD-AS1过表达,P-gp过表达(P<0.05);抑制lncRNA HOXD-AS1表达后OS细胞中P-gp表达下调(P<0.05);OS细胞中抑制lncRNA HOXD-AS1表达后细胞增殖转移能力明显减弱(P<0.05),凋亡增加(P<0.05),化疗耐药减弱(P<0.05);而过表达P-gp后恶性表型明显恢复(P<0.05)。结论:LncRNA HOXD-AS1可通过调控P-gp的表达而调节OS细胞的增殖、凋亡和化疗耐药,参与OS的发生发展。 Objective:To research the role and mechanism of lncRNA HOXD-AS1 in proliferation,apoptosis and doxorubicin chemotherapy-resistant of osteosarcoma(OS)cells.Methods:The expression of P-gp(P-glycoprotein)and lncRNA HOXD-AS1 in normal osteoblast,OS and OS/DOX(doxorubicin chemotherapy-resistant)cells were detected by real-time fluorescent quantitative PCR.The expression of P-gp in OS cells after down-regulted lncRNA HOXD-AS1 were examined by RT-PCR and Western-blot.The changes in abilities of proliferation in OS cells after inihibited lncRNA HOXD-AS1 and overexpressed P-gp were examined by CCK-8.Annexin V/PI staining was used to detect the apoptotic rates of each group at 48 h after transfection.Cell viability were detected after transfection and culture with doxorubicin.Results:LncRNA HOXD-AS1 was over-expression in OS cells and the expression levels of P-gp in OS cells were significantly increased as compared with those in normal osteoblast cells(P<0.05 for both).Transfection with siRNA of lncRNA HOXD-AS1 down-regulated P-gp expression(P<0.05).After inhibiting the expression of lncRNA HOXD-AS1 in OS cells,the cell proliferation ability was significantly weakened(P<0.05),apoptosis was increased(P<0.05),and chemotherapy resistance was weakened(P<0.05).And the overexpression of P-gp can reversed this phenomenon(P<0.05).Conclusion:LncRNA HOXD-AS1 can regulate the proliferation,apoptosis and doxorubicin chemotherapy-resistant of OS cells by regulating the expression of P-gp,and adjust the occurrence and development of OS.
作者 敖阳 张勇 AO Yang;ZHANG Yong(No.1 Orthopaedic Ward of Fuxin Chinese Traditional Medicine Hospital,Liaoning Fuxin 123000,China)
出处 《现代肿瘤医学》 CAS 北大核心 2021年第18期3155-3160,共6页 Journal of Modern Oncology
基金 辽宁省自然科学基金(编号:201601114)。
关键词 骨肉瘤 长链非编码RNA HOXD-AS1 P-GP 细胞增殖 细胞凋亡 化疗耐药 osteosarcoma long non-coding RNA HOXD-AS1 P-gp proliferation apoptosis chemotherapy-resistant
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