摘要
目的探讨长链非编码RNA RMRP(lncRNA-RMRP)对胶质瘤替莫唑胺(TMZ)耐药的影响,并检测lncRNA-RMRP在胶质瘤细胞系(U87、U118)TMZ耐药细胞株(U87TR、U118TR)和原发胶质瘤组织、复发胶质瘤组织中的表达情况。方法用实时荧光定量聚合酶链式反应(RT-q PCR)检测lncRNA-RMRP在U87、U87TR、U118、U118TR和胶质瘤组织、复发胶质瘤组织中的表达情况。U87TR、U118TR分别转染lncRNA-RMRP,并分为U87TR-RMRP组、U87TRNC组、U118TR-RMRP组和U118TR-NC组。用CCK-8方法检测400μmol·L^(-1)TMZ处理0,24,48,72,96 h后,U87、U87TR、U118、U118TR细胞的活力。用Western Blotting检测JAK2/STAT3相关蛋白的表达。结果 LncRNARMRP在U87、U87TR中表达量分别为(1.03±0.21)和(0.12±0.07),在U118、U118TR中表达量分别为(1.07±0.32)和(0.43±0.15),在复发胶质瘤组织和原发胶质瘤组织中表达量分别为(1.36±0.78)和(3.41±1.61),差异均有统计学意义(均P<0.01)。CCK-8检测结果显示,U87和U87TR的细胞增殖力分别为(2.62±0.22)和(4.01±0.12),U118和U118TR的细胞增殖力分别为(2.45±0.09)和(4.47±0.09),差异均有统计学意义(均P<0.01)。U87和U87TR的半抑制浓度(IC_(50))分别为(99.33±9.02)和(253.30±15.28)μmol·L^(-1),U118和U118TR的IC_(50)分别为(100.00±10.00)和(233.30±15.28)μmol·L^(-1),差异均有统计学意义(均P<0.01)。CCK-8检测结果显示,U87TR-RMRP和U87TR-NC的细胞增殖力分别为(2.18±0.14)和(4.36±0.13),U118TR-RMRP和U118TR-NC的细胞增殖力分别为(3.25±0.21)和(4.29±0.11),差异均有统计学意义(均P<0.01)。U87TRRMRP组和U87TR-NC组的TMZ IC_(50)分别为(142.30±6.81)和(283.30±15.28)μmol·L^(-1),U118TR-RMRP组和U118TR-NC组的TMZ IC_(50)分别为(189.70±16.01)和(300.30±10.02)μmol·L^(-1),差异均有统计学意义(均P<0.01)。U87TR-RMRP组和U87TR-NC组的p-JAK2分别为(1.02±0.23)和(0.49±0.13),p-STAT3分别为(1.02±0.0.21)和(0.28±0.07),差异均有统计学意义(均P<0.05)。U118TR-RMRP和U87TR-NC组的p-JAK2分别为(1.0
Objective To investigate the effect of long noncoding RNA RMRP（ lncRNA-RMRP） on the temozolomide（ TMZ） resistance ofglioma cells and to detect the expression of lncRNA-RMRP in regular glioma cells（ U87,U118） and TMZ-resistant glioma cells（ U87 TR,U118TR）,primary glioma tissues and relapsed glioma tissues. Methods Real-time quantitative PCR was used to detect the expression of lncRNA-RMRP in cell lines（ U87,U118,U87TR,U118TR）,primary and relapsed glioma tissues. U87TR and U118TR cells were transfected with lncRNA-RMRP. The experiment was divided into four groups： U87TR-RMRP,U87TR-NC,U118TR-RMRP and U118TR-NC. The viability of U87,U118,U87 TR and U118TR cells was tested by CCK-8 assay at 0,24,48,72 and 96 hours after TMZ（ 400μmol·L^-1） treatment. 50% inhibiting concentration（ IC50） was used to evaluate the resistance to TMZ in each treatment group. The expression of JAK2/STAT3 protein in each group was detected by Western blot. Results Real-time quantitative PCR showed that the expressions of LncRNA-RMRP were U87（ 1. 03 ± 0. 21） vs. U87TR（ 0. 12 ± 0. 07）,U118（ 1. 07 ± 0. 32） vs. U118TR（ 0. 43 ± 0. 15）,relapsed glioma tissues（ 1. 36 ± 0. 78） vs. primary glioma tissues（ 3. 41 ± 1. 61）,and the difference were statistically significant（ all P 0. 01）. The results of CCK-8 assay showed that the proliferative ability of U87 and U87TR were（ 2. 62 ± 0. 22） and（ 4. 01 ± 0. 12）,U118 and U118TR were（ 2. 45 ± 0. 09） and（ 4. 47 ± 0. 09）,the differences were statistically significant（ all P〈0. 01）.IC50 of U87 and U87TR were（ 99. 33 ± 9. 02） and（ 253. 30 ± 15. 28） μmol·L^-1,which in U118 and U118TR were（ 100. 00 ± 10. 00） and（ 233. 30 ± 15. 28） μmol·L^-1,the differences were statistically significant（ all P 0. 01）.The results of CCK-8 assay showed that the proliferative ability of U87TR-RMRP and U87TR-NC were（ 2. 18 ± 0. 14） and（ 4. 36 ± 0. 13）,U118TR-RMRP and U118TR-NC were（ 3. 25 ± 0. 21） and（
作者
马飞
李令兴
李琳
姜志平
高培平
MA Fei;LI Lin;LI Ling-xinglb;JIANG Zhi-ping;GAO Pei-ping(a Pharmacy Intravenous Admixture Services;b.Department of Cardiovascular Medicine,Taian City Central Hospital,Taian 271000,Shandong Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2018年第19期2320-2323,共4页
The Chinese Journal of Clinical Pharmacology
基金
中国医药教育协会基金资助项目(2016SKT-M030)
