PIM1在砷致细胞恶性转化过程中的作用
首发时间:2023-03-30
摘要:【目的】探究在砷致人皮肤角质形成细胞(HaCaT)恶性转化中PIM1的变化情况及其作用,为砷致癌机制的深入研究提供必要的基础资料。【方法】使用qRT-PCR方法检测经0.0或1.0 μM亚砷酸钠处理至35代过程中的各关键代数(0、1、7、14、21、28、35)的HaCaT细胞的PIM1、NRF2 mRNA的变化情况;Western Blot方法检测各关键代数的PIM1、NRF2蛋白的变化情况;使用PIM1 siRNA处理染毒至35代的HaCaT细胞,Western Blot检测PIM1、NRF2的蛋白变化情况;同时检测细胞倍增时间、细胞迁移率、软琼脂克隆形成率的变化情况。【结果】在砷致细胞恶性转化过程中PIM1、NRF2的mRNA水平以及蛋白水平逐渐上升,与对照组相比有统计学差异(P<0.05);使用PIM1 siRNA转染T-HaCaT细胞后,PIM1与NRF2蛋白水平均显著降低(P<0.05),且细胞的倍增时间显著上升、细胞迁移率与软琼脂克隆形成率显著降低(P<0.05)。【结论】本实验研究结果表明,在砷致细胞恶性转化过程中PIM1蛋白表达水平持续升高,且可能通过调控NRF2来促进细胞恶性转化。
关键词: 砷 亚砷酸钠 人皮肤角质形成细胞 恶性转化 PIM1
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The role of PIM1 in the malignant transformation of HaCaT cells induced by arsenite
Abstract:[Objective]To investigate the changes of PIM1 and its role in the malignant transformation of human keratinocytes (HaCaT) induced by arsenic, and to provide the necessary basic information for the in-depth study of the mechanism of arsenic carcinogenesis.[Method]HaCaT cells treated with 0.0 or 1.0 μM sodium arsenite up to 35 generations were examined by qRT-PCR for changes in PIM1, NRF2 mRNA at each key passage (0, 1, 7, 14, 21, 28, 35); Western blot method was used to detect changes in PIM1, NRF2 mRNA in each key passage; Western Blot to detect the protein changes of PIM1 and NRF2 in HaCaT cells stained with PIM1 siRNA up to 35 passages; and also to detect the changes of cell ploidy time, cell migration rate and soft agar clone formation rate.[Results]During the malignant transformation of arsenic-induced cells the mRNA levels of PIM1 and NRF2 as well as the protein levels increased gradually with the increase of arsenic staining time, and there was a statistical difference compared with the control group (P<0.05); after transfection of T-HaCaT cells with PIM1 siRNA, the protein levels of both PIM1 and NRF2 decreased significantly (P<0.05), and the cell doubling time was significantly increased, cell migration rate and soft agar clony formation rate were significantly decreased (P<0.05).[Conclusion]The results of the present experimental study showed that the expression level of PIM1 protein was consistently elevated during the malignant transformation of arsenic-induced cells, and it may promote the malignant transformation of cells through the regulation of NRF2.
Keywords: Arsenic Sodium arsenite Human keratinocytes Malignant transformation PIM1
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