CAJ | 학술논문

目的：观察脑胶质瘤组织、U251细胞中miR-7、EGFR/PI3K信号通路相关基因蛋白（ EGFR、PI3K和AKT2）的表达关系。方法42例脑胶质瘤患者，手术切除取肿瘤组织，其中8例选取癌旁正常脑组织。培养脑胶质瘤U251细胞，分为空白对照组（仅加入脂质体）、无义序列组（加入无义序列和脂质体）、miR-7转染组（转染miR-7基因片段）。采用RT-PCR法检测脑胶质瘤组织、各组U251细胞中的miR-7；免疫组化法检测胶质瘤组织中的EGFR、PI3K和AKT蛋白，RT-PCR和Western blotting法分别检测各组U251细胞EGFR、PI3K、AKT2 mRNA和蛋白。结果脑胶质瘤和正常脑组织中的miR-7 mRNA表达分别为1．47±0．27和3．47±0．15，两者比较，P＜0．05。随着脑胶质瘤WHO级别的增加，miR-7表达逐渐减少，EGFR、PI3K和AKT2表达增加（ P均＜0．05）。脑胶质瘤组织中miR-7与EGFR、PI3K、pAKT表达呈负相关（r分别为-0．754、-0．528、-0．467，P均＜0．05）。 U251细胞转染miR-7后EGFR、PI3K和AKT表达降低（P均＜0．05）。结论脑胶质瘤组织中miR-7表达减少，与EGFR/PI3K通路成员表达呈负相关；U251细胞转染miR-7后EGFR、PI3K和AKT表达降低。 EGFR/PI3K通路可能是miR-7调控胶质瘤发生发展的重要机制之一。
목적：관찰뇌효질류조직、U251세포중miR-7、EGFR/PI3K신호통로상관기인단백（ EGFR、PI3K화AKT2）적표체관계。방법42례뇌효질류환자，수술절제취종류조직，기중8례선취암방정상뇌조직。배양뇌효질류U251세포，분위공백대조조（부가입지질체）、무의서렬조（가입무의서렬화지질체）、miR-7전염조（전염miR-7기인편단）。채용RT-PCR법검측뇌효질류조직、각조U251세포중적miR-7；면역조화법검측효질류조직중적EGFR、PI3K화AKT단백，RT-PCR화Western blotting법분별검측각조U251세포EGFR、PI3K、AKT2 mRNA화단백。결과뇌효질류화정상뇌조직중적miR-7 mRNA표체분별위1．47±0．27화3．47±0．15，량자비교，P＜0．05。수착뇌효질류WHO급별적증가，miR-7표체축점감소，EGFR、PI3K화AKT2표체증가（ P균＜0．05）。뇌효질류조직중miR-7여EGFR、PI3K、pAKT표체정부상관（r분별위-0．754、-0．528、-0．467，P균＜0．05）。 U251세포전염miR-7후EGFR、PI3K화AKT표체강저（P균＜0．05）。결론뇌효질류조직중miR-7표체감소，여EGFR/PI3K통로성원표체정부상관；U251세포전염miR-7후EGFR、PI3K화AKT표체강저。 EGFR/PI3K통로가능시miR-7조공효질류발생발전적중요궤제지일。
Objective To explore the correlation between miR-7 and expression of EGFR/PI3K signal pathway relat-ed protein (EGFR, PI3K and AKT2) in brain glioma tissues and U251 cells.Methods The tumor tissues were obtained from 42 patients with brain glioma by excision , including 8 cases of tumor adjacent normal brain tissue .U251 glioma cells were cultured , and then were divided into the blank control group ( only with liposomes ) , nonsense sequence group ( non-sense sequence and liposomes ) and miR-7 transfection group ( transfection of miR-7 gene fragments ) .The expression of miR-7 in the glioma tissues and U251 cells was determined by RT-PCR.The expression of EGFR , PI3K and AKT2 protein in glioma tissues was detected by immunohistochemitry , and RT-PCR and Western blotting were used to detect the mRNA and protein expression of EGFR , PI3K and AKT2 in the U251 cells.Results The mRNA expression of miR-7 in the glio-ma and normal brain tissues were 1.47 ±0.27 and 3.47 ±0.15, respectively (P<0.05).With the increase of pathological grades of glioma, the expression of miR-7 decreased, the EGFR, PI3K and AKT expression increased (all P<0.05).The expression of miR-7 was negatively correlated with EGFR , PI3K and pAKT expression in the glioma tissues (r=-0.754,-0.528 and -0.467, respectively;all P<0.05).The EGFR, PI3K and AKT expression was significantly reduced in the U251 cells after being transfected with miR-7 (all P<0.05).Conclusions The expression of miR-7 in the glioma tissues is down-regulated, which is negatively correlated with the EGFR/PI3K pathway related protein.The expression of EGFR, PI3K and AKT is decreased in the U251 cells after being transfected with miR-7.The EGFR/PI3K pathway may be one of the important mechanisms of miR-7 in regulating the occurrence and development of gliomas .