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目的转录激活因子5(Activating transcription factor 5,ATF5)是一个新的与肿瘤细胞分化、增殖及凋亡密切相关的因子。文中检测胰腺癌SW1990细胞中ATF5的表达,并进一步研究紫杉醇(Paclitaxel,PTX)诱导的人胰腺癌SW1990细胞凋亡的水平,及其与ATF5、Bax表达水平的相关性。方法 RT-PCR检测未经药物处理的SW1990细胞中ATF5 mRNA表达水平;以不同浓度的紫杉醇(0、6.25、12.5、25、50、100、200 nmol/L)作用SW1990细胞不同时间(0、6、12、24、48 h),噻唑蓝(MTT)比色法检测细胞增殖情况;用100 nmol/L紫杉醇作用SW1990细胞不同时间(0、12、24、48 h)后,观察形态学变化并用Annexin V/7-AAD双染法,流式细胞仪检测细胞凋亡情况;采用RT-PCR检测紫杉醇作用不同时间后ATF5、Bax的mRNA表达变化。结果胰腺癌SW1990细胞中表达ATF5。紫杉醇抑制胰腺癌SW1990细胞的增殖,并在一定范围内呈剂量、时间相关性。随着紫杉醇作用时间的增加,流式细胞仪检测凋亡率显著提高。半定量RT-PCR检测结果显示:ATF5、Bax mRNA表达均明显增强,且两者之间存在相关性(r=0.916,P<0.05)。结论 ATF5在胰腺癌SW1990细胞中高表达,并且在紫杉醇诱导的细胞凋亡过程中,表达量进一步上升,其变化趋势与Bax基因相似。提示ATF5参与紫杉醇诱导的细胞凋亡,并可能与Bax的凋亡途径存在相关性。
Activating transcription factor 5 (ATF5) is a new factor closely related to the differentiation, proliferation and apoptosis of tumor cells. The expression of ATF5 in pancreatic cancer SW1990 cells was detected and the apoptosis of human pancreatic cancer SW1990 cells induced by paclitaxel (PTX) and its correlation with the expression levels of ATF5 and Bax were further investigated. Methods RT-PCR was used to detect the expression of ATF5 mRNA in untreated SW1990 cells. SW1990 cells were treated with different concentrations of paclitaxel (0, 6.25, 12.5, 25, 50, 100, 200 nmol/L) for different time (0, 6). , 12, 24, 48 h), MTT colorimetric assay was used to detect cell proliferation; SW1990 cells were treated with 100 nmol/L paclitaxel for different time (0, 12, 24, 48 h) and observed for morphological changes. The Annexin V/7-AAD double staining method was used to detect the apoptosis of the cells by flow cytometry. RT-PCR was used to detect the changes of ATF5 and Bax mRNA expression after paclitaxel exposure. Results ATF5 was expressed in pancreatic cancer SW1990 cells. Paclitaxel inhibited the proliferation of pancreatic cancer SW1990 cells and showed a dose- and time-dependent relationship within a certain range. With the increase of paclitaxel action time, the apoptosis rate was significantly increased by flow cytometry. The results of semi-quantitative RT-PCR showed that the expression of ATF5 and Bax mRNA was significantly increased and there was a correlation between them (r=0.916, P<0.05). Conclusion ATF5 is highly expressed in pancreatic cancer SW1990 cells, and the expression level of ATF5 is further increased during paclitaxel-induced apoptosis. The change tendency is similar to that of Bax gene. It suggests that ATF5 is involved in paclitaxel-induced apoptosis and may be related to the apoptotic pathway of Bax.