人胶质瘤细胞及组织中miRNA-221、miRNA-222表达变化及意义

Expression change and significance of miRNA-221/222 in glioma cell and tissue

目的观察人胶质瘤细胞及组织中miRNA-221、miRNA-222的表达变化,探讨其与肿瘤放射敏感性的相关性。方法根据Trizol试剂盒说明书,从体外培养的人胶质瘤细胞系U87、U251、A172、LN229和60例胶质瘤组织(手术切除获得)中提取其总RNA,采用RT-PCR法测定miRNA-221及miRNA-222,以正常脑组织为对照。60例胶质瘤患者术后均予以放疗,治疗后MRI检查随访,分析胶质瘤患者的放疗疗效。采用克隆形成实验测算X线照射后(照射剂量为2 Gy)的胶质瘤细胞存活率(SF),以胃黏膜上皮细胞(ges)为对照。结果与对照组织及细胞相比,胶质瘤组织及细胞中miRNA-221、miRNA-222呈高表达(P<0.05),其中高度恶性胶质瘤组织中miRNA-221、miRNA-222均明显高于低度恶性者(P<0.05)。X线照射后胶质瘤细胞SF明显高于ges(P均<0.05),胶质瘤各细胞中miRNA-221、miRNA-222表达量与X线照射后胶质瘤SF呈正相关(r=0.673、0.696,P均<0.01)。胶质瘤组织中miRNA-221、miRNA-222高表达的患者较低表达者生存时间短(P均<0.05)。结论人胶质瘤细胞及组织中miRNA-221、miRNA-222均呈高表达。miRNA-221、miRNA-222表达量与胶质瘤细胞的放射抗性呈正相关关系,胶质瘤组织中miRNA-221、miRNA-222表达量与患者生存时间呈负相关关系。

Objective To observe the expression changes of miRNA-221, miRNA-222 in human glioma cell and tissue, of which correlation with the radiosensitivity of human glioma was discussed. Methods The total RNA was extracted from in-vitro glioma cell lines U87, U251, A172, LN229 and 60 cases of glioma tissue （collected through operations） according to Trizol kit. The expression of miRNA-221 and miRNA-222 were detected with real-time PCR, and normal brain tissue was control sample. 60 cases of glioma patients was treated with radiotherapy after surgery. Glioma patients were follown-up and the radiotherapy in gliomas were analized by MRI examination. Glioma cell survival rate after X-ray irradiation （ irradiation dose of 2 Gy） was estimated by cloning experiments, with gastric mucosal epithelium cells as control. Results Compared with the control group, miRNA-221, miRNA-222 in glioma cell lines and tissue were highly expressed （all P 〈 0.05 ） . The expression of miRNA-221, miRNA-222 in high grade glioma was significantly higher than the low grade（ all P 〈 0.05 ）. The SF of glioma cell was significantly higher than that of ges after irradiation （ all P 〈 0.05 ）. MiRNA-221 and miRNA-222 expression in glioma cell was positively correlated with glioma SF after X-ray radiation（ r = 0. 673,0.696, all P 〈 0.05）. Glioma patients with miRNA-221, miRNA-222 highly expressed had a short survival time than the patients with miRNA-221, miRNA-222 low expression （ all P 〈 0.05）. Conclusions MiRNA-221 and miRNA-222 highly express in glioma cell and tissue, their expression was positively correlated with the radiation resistance of glioma cell, and their expression in glioma tissue are negatively correlated with the patients＇ survival time.