Pin1对内质网应激下肝癌HepG2细胞增殖和凋亡的影响

江铭婷; 黄晶; 郑淑萍

doi:10.3971/j.issn.1000-8578.2022.21.1097

Pin1对内质网应激下肝癌HepG2细胞增殖和凋亡的影响

Effects of Pin1 on Proliferation and Apoptosis of HepG2 Cells Under Endoplasmic Reticulum Stress

摘要

摘要:
目的探讨内质网应激下HepG2细胞中Pin1蛋白的表达情况及其对细胞增殖和凋亡的影响。
方法衣霉素（TM）分别作用于正常肝细胞THLE3和肝癌细胞HepG2（THLE3细胞TM组和HepG2细胞TM组）、全反式维甲酸（ATRA）单独作用于HepG2细胞（ATRA组）、TM和ATRA共同作用于HepG2细胞（TM+ATRA组）及DMSO处理的对照组细胞。48 h后，Western blot检测Bip和Pin1蛋白的表达水平，CCK-8试剂检测细胞的增殖情况，流式细胞术分析细胞的凋亡情况。
结果 TM处理的THLE3和HepG2细胞中Bip蛋白表达增加，说明TM有效诱导细胞发生内质网应激；与DMSO组相比，THLE3细胞TM组中Pin1的蛋白水平随着TM浓度的升高而降低（P < 0.001）。HepG2细胞TM+ATRA组中Pin1蛋白水平随着TM浓度的升高而降低（P < 0.01）。HepG2细胞TM组细胞增殖抑制率仅（29.33±4.73）%，明显低于THLE3细胞TM组（（60.33±2.08）%）（P < 0.001），但HepG2细胞TM+ATRA组细胞增殖抑制率显著提高至（60.33±6.03）%。与DMSO组相比，THLE3细胞TM组凋亡率显著升高（（22.25±0.78）% vs.（3.57±0.31）%, P < 0.01）。与DMSO组（5.10±1.00）%相比，HepG2细胞ATRA组凋亡率只有（10.03±0.49）%（P < 0.05），但TM+ATRA组凋亡率显著增加至（23.70±0.75）%（P < 0.01）。
结论 HepG2细胞通过维持Pin1蛋白水平抵制ERS诱导的细胞凋亡，降低Pin1蛋白水平能够抑制细胞增殖、促进细胞凋亡。

Abstract:
Objective To investigate the expression of Pin1 protein in HepG2 cells under endoplasmic reticulum stress (ERS) and its effect on cell proliferation and apoptosis.
Methods The THLE3 cells were treated with tunicamycin (TM) (TM group) or DMSO (DMSO group) for 48h. The HepG2 cells were treated with TM (TM group), ATRA(ATRA group), TM+ATRA (TM+ATRA group) or DMSO (DMSO group) for 48h. The protein levels of Bip and Pin1 were detected by Western blot, cell proliferation was detected by CCK-8 assay, and cell apoptosis was detected by flow cytometry.
Results The expression of Bip both increased in THLE3 and HepG2 cells treated with TM, indicated that TM effectively induced ERS in cells. Compared with the DMSO group, the protein level of Pin1 in THLE3 cells in TM group was decreased with the increasing of TM concentration (P < 0.001). In TM+ATRA group, with the increasing of TM concentration, the expression of Pin1 was decreased in HepG2 cells (P < 0.01). The inhibitory rates was (29.33±4.73)% in HepG2 cells in TM group, significantly lower than (60.33±2.08)% in THLE3 cells in TM group (P < 0.001). In TM+ATRA group, the growth inhibition rates was increased to (60.33±6.03)% in HepG2 cells. The apoptosis rate of THLE3 cells in TM group was (22.25±0.78)%, significantly higher than (3.57±0.31)% in DMSO group (P < 0.01). The apoptosis rates of HepG2 cells in ATRA group and TM+ATRA group were significantly higher than that in the DMSO group ((10.03±0.49)% vs. (5.10±1.00)%, P < 0.05 and (23.70±0.75)% vs. (5.10±1.00)%, P < 0.01).
Conclusions HepG2 cells resist ERS-induced apoptosis by maintaining Pin1 protein level. Reducing the protein level of Pin1 can significantly inhibit the cell proliferation and induce apoptosis.

HTML全文

参考文献(19)

施引文献

资源附件(0)