CAJ | 학술논문

探讨造血生长因子GM-CSF/IL-3融合蛋白、GM-CSF及IL-3抗γ射线辐射诱导Tf-1细胞凋亡的分子机制.采用荧光强度比色法检测辐射后Tf-1细胞内Caspase-3活性;RT-PCR半定量及免疫组化实验方法分别观察Bcl-2、Caspase-3 mRNA及蛋白表达水平的变化.GM-CSF/IL-3融合蛋白、GM-CSF、IL-3以及GM-CSF与IL-3合用均可使辐射后Tf-1细胞内Caspase-3活性明显降低,其活性是不加入任何细胞因子对照组的15.71%-43.65%.在Tf-1细胞辐射后48 h时,各细胞因子组Bcl-2 mRNA及蛋白的表达水平较对照组明显提高;其中GM-CSF/IL-3融合蛋白10μg/L组在辐射后24h时观察到Bcl-2 mRNA表达增强.细胞辐射后48h时,GM-CSF/IL-3融合蛋白10μg/L、100μg/L及IL-3 10μg/L组Caspase-3mRNA的表达明显增强;辐射后48h时,各细胞因子组均观察到Caspase-3蛋白表达增多.三种细胞因子可通过明显抑制Caspase-3活性、促进Bcl-2、Caspase-3 mRNA、蛋白质的表达以及抑制Caspase-3酶原激活而发挥抗辐射诱导的Tf-1细胞凋亡.
탐토조혈생장인자GM-CSF/IL-3융합단백、GM-CSF급IL-3항γ사선복사유도Tf-1세포조망적분자궤제.채용형광강도비색법검측복사후Tf-1세포내Caspase-3활성;RT-PCR반정량급면역조화실험방법분별관찰Bcl-2、Caspase-3 mRNA급단백표체수평적변화.GM-CSF/IL-3융합단백、GM-CSF、IL-3이급GM-CSF여IL-3합용균가사복사후Tf-1세포내Caspase-3활성명현강저,기활성시불가입임하세포인자대조조적15.71%-43.65%.재Tf-1세포복사후48 h시,각세포인자조Bcl-2 mRNA급단백적표체수평교대조조명현제고;기중GM-CSF/IL-3융합단백10μg/L조재복사후24h시관찰도Bcl-2 mRNA표체증강.세포복사후48h시,GM-CSF/IL-3융합단백10μg/L、100μg/L급IL-3 10μg/L조Caspase-3mRNA적표체명현증강;복사후48h시,각세포인자조균관찰도Caspase-3단백표체증다.삼충세포인자가통과명현억제Caspase-3활성、촉진Bcl-2、Caspase-3 mRNA、단백질적표체이급억제Caspase-3매원격활이발휘항복사유도적Tf-1세포조망.
To investigate the molecular mechanisms of the anti-apoptosis effects of hematopoietic growth factors GM-CSF/IL-3 fusion protein, GM-CSF and IL-3 in Tf-1 cells induced by γ-irradiation. Caspase-3 activity was detected using fluorescence chromatometry. RT-PCR semiquantitative analysis was used to identify the expression changes of Bcl-2 and Caspase-3 mRNA, and an immunohistochemical technique was used to examine the protein expression in irradiated Tf-1 cells. The Caspase-3 activity in irradiated Tf-1 cells was markedly reduced to 15.71 %-43.65 % in GM-CSF/IL-3 fusion protein, GM-CSF, IL-3, and the combination of GM-CSF and IL-3-treated groups compared to those in the control groups without any growth factor. Bcl-2 mRNA and its protein expression were significantly increased by the above cytokines after incubation with the cells for 48 h after irradiation, while the enhancement of the Bcl-2 mRNA level by GM-CSF / IL-3 fusion protein (10 μg / L) began from 24 h. After 48h of incubation after irradiation, the expression of Caspase-3 mRNA was dramatically augmented by GM-CSF / IL-3 fusion protein at 10 μg / L, 100 μg / L and IL-3 10 μg / L, as was the corresponding Caspase-3 protein expression by the addition of each cytokine. The cytokines exerted the anti-apoptosis effects in irradiated Tf-1 cells through the up-regulation of Bcl-2 and Caspase-3 expression, and the inhibition of Caspase-3 activity.