标题：干扰MGMT的人胶质瘤U251细胞对替莫唑胺的敏感性变化

      作者：罗冬冬 1，彭彪 1，罗爱萍 2，胡骕 1，赵海林 1，李丹 1    (广州医科大学附属肿瘤医院神经外科 1、肝胆外科 2，广东 广州 510095)

      卷次： 2018年29卷13期

      【摘要】 目的 观察干扰O6-甲基鸟嘌呤DNA甲基转移酶(MGMT)的人胶质瘤U251细胞对替莫唑胺(TMZ)的敏感性变化。方法 培养人胶质瘤U251细胞，将U251细胞经16 mg/L替莫唑胺处理后分别转染 siRNA-MGMT与 siRNA-control，转染 siRNA-MGMT质粒组为观察组，转染 siRNA-control质粒组为对照组，不转染任何序列组为单药组，共培养48 h后收获细胞。qRT-PCR法检测三组细胞中MGMT的mRNA表达改变，MTT法观察各组U251细胞增殖能力，Transwell实验检测各组U251细胞侵袭能力。结果 qRT-PCR检测结果显示，观察组U251细胞中MGMT的mRNA表达量为(0.285±0.031)，与对照组(1.104±0.096)和单药组(0.974±0.083)比较均明显下降，差异均有统计学意义(P<0.05)，而对照组与单药组比较差异无统计学意义(P>0.05)；MTT检测结果显示，观察组U251细胞的OD值为(0.392±0.014)，与对照组(0.561±0.021)和单药组(0.530±0.019)比较均明显下降，差异均有统计学意义(P<0.05)，而对照组与单药组比较差异无统计学意义(P>0.05)；Transwell检测结果显示，观察组U251细胞穿过基质胶的数量为(64.27±7.04)个，与对照组[(128.14±14.37)个]和单药组[(119.63±10.92)个]比较均明显减少，差异均有统计学意义(P<0.05)，而对照组与单药组比较差异无统计学意义(P>0.05)。结论 干扰MGMT的人胶质瘤U251细胞对替莫唑胺的敏感性增强。
      【关键词】 人胶质瘤U251细胞；O6-甲基鸟嘌呤DNA甲基转移酶；替莫唑胺；化疗敏感性
      【中图分类号】 R739.41 【文献标识码】 A 【文章编号】 1003—6350（2018）13—1777—03

Sensitivity changes of human glioma U251 cells interfered with MGMT to temozolomide.
LUO Dong-dong 1, PENGBiao 1, LUO Ai-ping 2, HU Su 1, ZHAO Hai-lin 1, LI Dan 1. Department of Neurosurgery 1, Department of HepatobiliarySurgery 2, the Affiliated Tumor Hospital of Guangzhou Medical University, Guangzhou 510095, Guangdong, CHINA
【Abstract】 Objective To observe the sensitivity of glioma U251 cells interfered with O(6)-methylgua-nine-DNA methyltransferase (MGMT) to temozolomide (TMZ). Methods Human glioma U251 cells were cultured,and were transfected with siRNA-MGMT and siRNA-control after treated with 16 mg/L of TMZ. The transfected siR-NA-MGMT plasmid group was used as the observation group, and the siRNA-control plasmid group was used as thecontrol group. The single drug group was not transfected with any sequence, and all cells were harvested after co-culturefor 48 h. The expression of MGMT mRNA was detected by qRT-PCR. MTT ((3-[4,5-dimethylthiazol-2-yl]-2,5 di-phenyl tetrazolium bromide) assay was performed to detect the proliferation ability of U251 cells in each group. Tran-swell assay was used to detect the invasive ability of U251 cells in each group. Results qRT-PCR assay showed thatthe expression of MGMT mRNA in the observation group was (0.285 ± 0.031), which was significantly lower than(1.104±0.096) of control group and (0.974±0.083) of the single drug group (P<0.05). But there was no significant differ-ence between the control group and the single drug group (P>0.05). As shown by MTT assay, the OD value of U251cells in the observation group was (0.392±0.014), which was significantly lower than (0.561±0.021) in the control groupand (0.530±0.01) in the single drug group (P<0.05), with no singnificant difference between the last two groups (P>0.05).Transwell test results showed that the number of U251 cells in the observation group that passed through Matrigel was(64.27±7.04), which was significantly reduced compared with (128.14±14.37) in the control group and (119.63±10.92) inthe single drug group (P<0.05), but there was no significant difference between the control group and the single druggroup (P>0.05). Conclusion Interfered MGMT in human glioma U251 cells can increase the sensitivity to TMZ.
      【Key words】 Human glioma U251 cells; O(6)-methylguanine-DNA methyltransferase (MGMT); Temozolomide(TMZ); Sensitivitydoi:10.3969/j.issn.1003-6350.2018.13.001基金项目：广州医科大学青年科研项目(编号：2015C39)