ARHI基因与HuR蛋白在胰腺癌PANC-1细胞中的表达

杨红; 胡珊珊; 阮戈冲; 杨晓鸥; 钱家鸣

doi:10.3969/j.issn.1674-9081.2017.01.009

ARHI基因与HuR蛋白在胰腺癌PANC-1细胞中的表达

Expression of ARHI and HuR in PANC-1 Cells

摘要

摘要:
目的观察ARHI基因转染胰腺癌PANC-1细胞后对HuR蛋白的影响。
方法采用脂质体法将表达ARHI基因的质粒pLNCX2-ARHI-EGFP和空载体pLNCX2-EGFP转染胰腺癌PANC-1细胞，细胞分为3组：实验组(pLNCX2-ARHI-EGFP)、空载体转染组(pLNCX2-EGFP)和PANC-1细胞空白对照组，用G418筛选稳定转染的细胞株；采用PCR和Western blot方法检测ARHI基因表达，采用Western blot方法检测HuR蛋白表达。
结果稳定转染ARHI基因组可检测到ARHI基因mRNA和蛋白的表达。根据图像灰度计算蛋白条带相对表达量，稳定转染ARHI基因组HuR蛋白表达(0.2226± 0.0644)较空载体对照转染组(0.4063 ± 0.0925，P=0.029)和PANC-1细胞空白对照组(0.4178 ± 0.1319，P=0.022)显著降低。
结论 ARHI基因可以导致胰腺癌细胞株中HuR蛋白表达降低。

Abstract:
Objective To investigate the effect of ARHI transfection on the expression of HuR protein in PANC-1 cells.
Methods Plasmids pLNCX2-ARHI-EGFP which express ARHI and empty plasmid pLNCX2-EGFP were used to transfect PANC-1 cells, which were divided into three groups: pLNCX2-ARHI-EGFP group, pLNCX2-EGFP group, and blank control.G418 was used to select stable transfected cell lines. ARHI gene expression was detected by PCR and Western blot, and the expression of HuR protein was detected by Western blot.
Results PANC-1 cells transfected with pLNCX2-ARHI-EGFP, the mRNA and protein expression of ARHI were detected. HuR protein expression appeared significantly decreased in PANC-1 cells transfected with pLNCX2-ARHI-EGFP(0.2226±0.0644) compared with the pLNCX2-EGFP-transfected cells (0.4063±0.0925, P=0.029) and PANC-1 blank control cells (0.4178±0.1319, P=0.022).
Conclusion ARHI gene in PANC-1 cells may reduce the expression of HuR protein.

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