摘要
目的探讨非编码RNA CTBP1-AS2通过调节O-6-甲基鸟嘌呤DNA甲基转移酶(MGMT)对高级别脑胶质瘤细胞替莫唑胺耐药的影响。方法选取2019年6月至2020年12月手术切除的脑胶质瘤样本,其中初发高级别胶质瘤20例(高级别初发组),复发高级别胶质瘤(手术切除和替莫唑胺治疗后复发)20例(高级别复发组),另选取颅脑损伤减压术获取的正常脑组织20例(对照组)。用RT-PCR法检测CTBP1-AS2相对表达量。分别用1、2、4和8μg·mL^(-1)替莫唑胺诱导胶质瘤U251细胞,构建U251/替莫唑胺耐药(U251/TR)细胞系;用慢病毒转染构建稳定干扰CTBP1-AS2的U251/TR(U251/TR-CTBP1-AS2)细胞系。检测U251细胞、U251/TR细胞系、U251/TR-CTBP1-AS2细胞系的替莫唑胺IC50值。RT-PCR法检测细胞CTBP1-AS2表达;Western blot方法检测MGMT蛋白表达水平。结果高级别复发组中CTBP1-AS2相对表达量为(0.34±0.10),较对照组(0.13±0.08)和高级别初发组(0.28±0.05)增高(均P<0.05)。U251细胞、U251/TR细胞系、U251/TR-CTBP1-AS2细胞系的IC50分别为(3.91±0.23)μg·mL^(-1)、(8.13±1.36)μg·mL^(-1)、(5.96±1.32)μg·mL^(-1);U251/TR细胞系的IC50高于U251细胞(P<0.05),U251/TR-CTBP1-AS2细胞系的IC50低于U251/TR细胞系(P<0.05)。U251/TR细胞系的CTBP1-AS2基因和MGMT蛋白表达量均高于U251细胞(均P<0.05)。U251/TR-CTBP1-AS2细胞系的CTBP1-AS2基因和MGMT蛋白表达量均低于U251/TR细胞系(均P<0.05)。结论CTBP1-AS2可能通过调节MGMT促进高级别胶质瘤细胞的替莫唑胺耐药。
Aim To explore the effect of non-coding RNA CTBP1-AS2 on temozolomide resistance of high-grade glioma cells by regulating O-6-methylguanine DNA methyltransferase(MGMT).Methods Selected tissue samples from our hospital from June 2019 to December 2020.Among them,there were 20 cases of newly developed high-grade gliomas,20 cases of recurrent high-grade gliomas(recurrence after surgical resection and temozolomide treatment).20 cases of normal brain tissues taken during internal decompression of craniocerebral injury were used as the control group.RT-PCR was used to detect the expression of CTBP1-AS2 in the tissues.Glioma U251 cells were induced to construct U251 temozolomide-resistant cell line(U251/TR)by using 1μg·mL^(-1),2μg·mL^(-1),4μg·mL^(-1),8μg·mL^(-1) temozolomide respectively.The U251/TR cell line(U251/TR-CTBP1-AS2)stably interfering with CTBP1-AS2 was constructed by lentivirus transfection.Temozolomide IC50 values of U251 cells,U251/TR cells and U251/TR-CTBP1-AS2 cells were detected.The expression of CTBP1-AS2 was detected by RT-PCR.The expression of MGMT protein was detected by Western-blot.Results The relative expression of CTBP1-AS2 in recurrent high-grade glioma tissue was(0.34±0.10),which was higher than that in normal control brain tissue(0.13±0.08)and newly-onset high-grade glioma tissue(0.28±0.05)(P<0.05).The IC50 of U251 cells,U251/TR cells,and U251/TR-CTBP1-AS2 cells are(3.91±0.23)μg·mL^(-1),(8.13±1.36)μg·mL^(-1),(5.96±1.32)μg·mL^(-1),respectively.The IC50 of U251/TR cells was higher than that of U251 cells(P<0.05),the IC50 of U251/TR-CTBP1-AS2 was lower than that of U251/TR cells(P<0.05).The expression levels of CTBP1-AS2 gene and MGMT protein in U251/TR cells were higher than U251 cells(P<0.05).The expression levels of CTBP1-AS2 gene and MGMT protein in U251/TRCTBP1-AS2 cells were lower than U251/TR cells(P<0.05).Conclusion CTBP1-AS2 may promote temozolomide resistance in high-grade glioma cells by regulating MGMT.
作者
赵鹏飞
刘旭
门帅
ZHAO Peng-fei;LIU Xu;MEN Shuai(Department of Neurosurgery,Nan Shi Hospital Affiliated to Henan University,Nanyang 473000,China)
出处
《中国临床神经科学》
2022年第5期514-518,共5页
Chinese Journal of Clinical Neurosciences
