LncRNA TTN-AS1调控骨肉瘤多药耐药细胞对阿霉素敏感性的功能及机制  

The function and mechanism of LncRNA TTN⁃AS1 regulating the sensitivity of multidrug resistant osteosarcoma cells to adriamycin

作　　者：李战鹏 刘鹏飞 李斌 LI Zhanpeng;LIU Pengfei;LI Bin(Department of Orthopaedics,Zhangjiakou First Hospital,Zhangjiakou,Hebei 075000,China)

机构地区：[1]张家口市第一医院骨科,河北张家口075000

出　　处：《热带医学杂志》2024年第8期1094-1099,1129,共7页Journal of Tropical Medicine

基　　金：河北省医学科学研究课题项目(20221885);张家口市科学技术研究与发展计划项目(1711042H)。

摘　　要：目的探究长链非编码RNA(LncRNA)TTN-AS1调控骨肉瘤细胞对阿霉素(ADR)敏感性的功能及机制。方法骨肉瘤细胞株购自中国科学院上海细胞库。实时荧光定量逆转录聚合酶链式反应(RT-qPCR)检测TTN-AS1在骨肉瘤细胞株MG63和多药耐药细胞MG63/Dox中的表达,MG63/Dox设置为Control组、si-TTN-AS1组、ADR组、si-TTN-AS1+ADR组、si-TTN-AS1+miR-134-5p inhibitor+ADR组、si-TTN-AS1+miR-134-5p inhibitor+si-MBTD1+ADR组,双荧光素酶报告基因实验检测TTN-AS1和miR-134-5p的靶向关系以及miR-134-5p和恶性脑肿瘤区包含蛋白1(MBTD1)的靶向关系,CCK8和流式细胞术检测各组细胞增殖和凋亡。结果与MG63细胞相比,MG63/Dox细胞中TTN-AS1相对表达水平显著升高,差异有统计学意义(t=6.43,P<0.05)。与si-TTN-AS1组相比,si-TTN-AS1+ADR组在24、48、72 h的MG63/Dox细胞增殖能力显著降低(t=4.16、5.06、5.72),与si-TTN-AS1组和ADR组相比,si-TTN-AS1+ADR组细胞凋亡显著升高(t=6.23、14.84),差异均有统计学意义(P均<0.05)。双荧光素酶报告基因结果显示,miR-134-5p为TTN-AS1靶基因,MBTD1为miR-134-5p靶基因。与si-TTN-AS1+ADR组相比,si-TTN-AS1+miR-134-5p inhibitor+ADR组在24、48、72 h的MG63/Dox细胞增殖能力显著升高(t=10.66、10.96、14.03),细胞凋亡显著降低(t=5.58),差异均有统计学意义(P均<0.05)。与si-TTN-AS1+miR-134-5p inhibitor+ADR组相比,si-TTN-AS1+miR-134-5p inhibitor+si-MBTD1+ADR组在24、48、72 h的MG63/Dox细胞增殖能力显著降低(t=5.78、7.85、8.38),细胞凋亡显著升高(t=7.96),差异均有统计学意义(P均<0.05)。结论TTN-AS1通过调控miR-134-5p/MBTD1轴,影响MG63/Dox对ADR的敏感性。Objective To investigate the function and mechanism of long non⁃coding RNA(LncRNA)TTN⁃AS1 in regulating the sensitivity of osteosarcoma cells to adriamycin(ADR).Methods Osteosarcoma cell lines were purchased from the Shanghai Cell Bank,Chinese Academy of Sciences.Reverse transcription⁃quantitative polymerase chain reaction(RT⁃qPCR)was used to detect the expression of TTN⁃AS1 in osteosarcoma cell line MG63 and multidrug⁃resistant cell line MG63/Dox.MG63/Dox was divided into Control group,si⁃TTN⁃AS1 group,ADR group,si⁃TTN⁃AS1+ADR group,si⁃TTN⁃AS1+miR⁃134⁃5p inhibitor+ADR group and si⁃TTN⁃AS1+miR⁃134⁃5p inhibitor+si⁃MBTD1+ADR group.The dual luciferase reporter gene experiment was used to detect the targeting relationship between TTN⁃AS1 and miR⁃134⁃5p,as well as the targeting relationship between miR⁃134⁃5p and malignant brain tumour domain containing 1(MBTD1).CCK8 and flow cytometry were used to detect cell proliferation and apoptosis in each group.Results The relative expression level of TTN⁃AS1 in MG63/Dox cells was significantly higher than that in MG63 cells(t=6.43,P<0.05).The proliferation ability of MG63/Dox cells in 24,48,72 h in the si⁃TTN⁃AS1+ADR group was significantly lower than that in the si⁃TTN⁃AS1 group(t=4.16,5.06,5.72),and cell apoptosis was significantly higher than that in the si⁃TTN⁃AS1 group and ADR group(t=6.23,14.84),the differences were statistically significant(all P<0.05).MiR⁃134⁃5p is the TTN⁃AS1 target gene,and MBTD1 is the miR⁃134⁃5p target gene.The proliferation ability of MG63/Dox cells in 24,48,72 h in the si⁃TTN⁃AS1+miR⁃134⁃5p inhibitor+ADR group was significantly higher than that in the si⁃TTN⁃AS1+ADR group(t=10.66,10.96,14.03),and cell apoptosis was significantly lower than that in the si⁃TTN⁃AS1+ADR group(t=5.58),the differences were statistically significant(all P<0.05).The proliferation ability of MG63/Dox cells in 24,48,72 h in the si⁃TTN⁃AS1+miR⁃134⁃5p inhibitor+si⁃MBTD1+ADR grou

关 键 词：TTN-AS1 长链非编码RNA 骨肉瘤 阿霉素 敏感性 

分 类 号：R738[医药卫生—肿瘤]