脑胶质瘤白细胞介素-6自分泌或旁分泌环路的研究

The study on the autocrine or paracrine interleukin-6/interleukin-6 receptor loop in gliomas

目的 研究白细胞介素 6(IL 6) /IL 6受体 (IL 6R)自分泌或旁分泌环路与脑胶质瘤的关系。方法 采用逆转录 聚合酶链反应 (RT PCR)方法检测IL 6和IL 6R基因在 4株脑胶质瘤细胞株、62例脑胶质瘤组织标本、5例良性脑膜瘤组织和 5例人正常脑胶质细胞中的表达 ;将IL 6单抗 (McAb)或IL 6RMcAb与脑胶质瘤细胞共同孵育 ,四甲基偶氮唑蓝 (MTT)比色法检测对细胞生长的抑制作用。结果 脑胶质瘤细胞株C6、9L、U 2 5 1和SHG44均有IL 6和IL 6R的表达 ,脑胶质瘤组织标本中 47例 (75 .81% )表达IL 6、5 2例 (83 .87% )表达IL 6R、41例 (66.13 % )同时表达IL 6和IL 6R ;良性脑膜瘤组织 4例表达IL 6,人脑正常胶质细胞仅有IL 6弱表达 ,两者均无IL 6R表达。IL 6McAb和IL 6RMcAb能够显著抑制脑胶质瘤细胞的增殖 ,并存在剂量依赖关系(P <0 .0 1)。结论 脑胶质瘤很可能存在IL 6/IL 6R自分泌或旁分泌环路 。

Objective To study the relationship between the interleukin-6 (IL-6)/IL-6 receptor (IL-6R) autocrine or paracrine loop and the malignant proliferation of glioma cells.Methods IL-6 and IL-6R gene was detected in adult normal astrocytes,benignant meningioma tissues,4 glioma cell lines and 62 human glioma tissue specimens with different malignant degrees.A complementary DNA copy of total RNA was synthesized and amplified with specific primers using the reverse transcription polymerase chain reaction (RT-PCR) method.Glioma cells,C6,9L,U251 and SHG44 were cultured with IL-6 McAb and IL-6R McAb in different concentrations.Then the proliferation of glioma cells was evaluated using MTT assay method.Results The experiments revealed that all of 4 glioma cell lines expressed IL-6 and IL-6R.IL-6 gene was found in 47 samples (75.81 %) and positive IL-6R gene in 52 samples (83.87 %) respectively.There were 41 (66.13 %) glioma tissue specimens that co-expressed IL-6 and IL-6R gene.Moreover,the positive rate and intensity of the expression of two genes was related to the malignant degree of gliomas.On the other hand,the expression of IL-6 was weakly positive in adult normal astrocytes and benignant meningioma tissues,but IL-6R was negative.Furthermore,it was discovered that both IL-6 McAb and IL-6R McAb could inhibit cell growth in a dose-dependent manner by blocking the IL-6/IL-6R autocrine or paracrine loop.Conclusion There maybe is autocrine or paracrine IL-6/IL-6R loop in gliomas,which could improve the glioma cell growth.It is possibly to develop anti-glioma biotherapy using IL-6McAb or/and IL-6RMcAb to inhibit the network of IL-6/IL-6R loop.