三维培养条件下电离辐射诱导适应性反应研究

目的研究在三维细胞培养条件下,电离辐射诱导人成纤维细胞发生适应性反应。方法采用人成纤维细胞进行三维条件培养,以60Co进行照射（诱导剂量D1为0.1Gy,攻击剂量D2为2Gy）,以Western blot检测生长相关基因p53和p21的表达,免疫荧光染色及共聚焦法观察形态学变化。结果三维培养条件下,细胞生长与二维（单层）培养形态学表现明显不同,细胞形态伸展,呈现伪足和多微丝状,细胞正常∶异常∶中间态比例为87.05%∶6.1%∶6.85%。经过2Gy（D2）照射后细胞明显收缩,伪足和微丝消失,与对照组比较,细胞变为正常∶异常∶中间态比例30.4%∶67.45%∶2.15%;预先给以0.1Gy即D1＋D2照射后则出现异常细胞数量减少,正常细胞有所增加,同时不典型的中间态细胞比例增加,细胞比例为正常∶异常∶中间态35.5%∶34.7%∶30.4%。结论人成纤维细胞在三维培养条件下,预先低剂量60Co照射可以降低随后高剂量照射引起的形态学改变,诱导产生适应性反应。

Ionizing radiation_induced adaptive response in human fibroblasts under 3_dimentional culture condition

Objective To observe the adaptive response induced by ionizing radiation in human fibroblasts under 3_dimentional culture condition.Methods Human fibroblasts were cultured under 3_D condition.Western blot was used to detect the expressions of p53 and p21.Immunofluroescence and confocal microscopy were used to detect the morphological changes.Results The structures of the cells under 2_D and 3_D conditions were fundamentally different,the cells under 3_D condition stretched out and formed microspikes,the proportions of normal, abnormal and in between were 87.05%,6.1% and 6.85%,respectively.After the treatment of 2 Gy(D₂) most of the fibroblasts shrinked and the microspikes disappeared,while the pre_tretment of 0.1 Gy(D₁) pulsing the challenging dose of 2 Gy(i.e.,D₁+D₂) caused the decrease of abnormal cells and the increase of the normal cells and in between(35.5%:34.7%:30.4%).Conclusions Under the 3_D conditions pretreatment with low dose radiation could attenuate the morphological changes induced by high dose.i.e. induced the adaptive response.