基于质粒的蛋白激酶B1特异性小干扰RNA和P53共表达协同抑制肝细胞癌细胞的增殖、迁移和侵袭

doi:10.16098/j.issn.0529-1356.2021.02.014

摘要/Abstract

摘要：

目的探讨蛋白激酶B1（Akt1）特异性小干扰RNA（siRNA）和p53共表达质粒对肝细胞癌（HCC）细胞增殖、迁移、侵袭、凋亡的影响。方法构建了一种共表达蛋白激酶B1（Akt1）特异性siRNA和野生型p53基因的双表达质粒(pSi-Akt1-P53)。将pSi-Akt1-P53共表达质粒转染至肝癌HepG2细胞，采用Real-time PCR法和免疫印迹法检测细胞中Akt1和P53表达被调控情况。然后将这种共表达质粒转染至HepG2细胞中，并同shAkt1质粒和P53质粒相比较，分别采用CCK-8和5-乙炔基-2’-脱氧尿苷（EdU）实验、划痕实验、Transwell小室法、流式细胞术检测其对HepG2细胞增殖、迁移、侵袭和凋亡的影响。结果 pSi-Akt1-P53共表达质粒转染至肝癌HepG2细胞后显著地减少HepG2细胞中Akt1蛋白的表达，同时显著增加P53蛋白的表达。与shAkt1质粒和P53质粒相比，pSi-Akt1-P53共表达质粒更显著地抑制HepG2细胞的增殖、迁移和侵袭能力，同时更显著地促进HepG2细胞的凋亡。结论 pSi-Akt1-P53共表达质粒能非常显著地抑制肝癌HepG2细胞的增殖、迁移和侵袭能力，有效促进肝癌HepG2细胞凋亡。

关键词: 肝细胞癌, 共表达质粒, 蛋白激酶B1, P53, 实时定量聚合酶链反应, 人

Abstract:

Objective To investigate the effect of the dual expression plasmid of protein kinase B1(Akt1)-specific siRNA and P53 on the proliferation, migration, invasion and apoptosis of hepatocellular carcinoma (HCC) cells. Methods We constructed a dual expression plasmid that co-expressed Akt1-specific siRNA and wild-type p53 gene (pSi-Akt1-P53).The dual expression plasmid pSi-Akt1-P53 was transfected into HepG2 cells of HCC,The expression of Akt1 and P53 was detected by Real-time PCR and Western blotting. Then, the dual expression plasmid was transfected into HepG2 cells, sh-Akt1 plasmid and P53 plasmid were used as control. The effects of the dual plasmid on the proliferation, migration, invasion and apoptosis of HepG2 cells were detected by CCK-8 and 5-ethynyl-2’-deoxyuridine(EdU) experiments, Wound scratch experiment, Transwell chamber experiment and flow cytometry,respectively. Results After the dual plasmid was transfected into HepG2 cells, the expression of Akt1 protein was significantly reduced and the expression of P53 protein was significantly increased in HepG2 cells. Compared with the shAkt1 and P53 plasmids, the dual expression plasmid pSi-Akt1-P53 significantly inhibited the proliferation、migration and invasion of HepG2 cells and significantly increased the apoptosis of HepG2 cells. Conclusion The dual expression plasmid pSi-Akt1-P53 can synergistically inhibit the proliferation, migration and invasion of HepG2 cells, significantly increased the apoptosis of HepG2 cells.

Key words: Hepatocellular carcinoma, Dual expression plasmid, Protein kinase B1, P53, Real-time PCR, Human

中图分类号:

R735.7

陈小龙万亚锋黄平. 基于质粒的蛋白激酶B1特异性小干扰RNA和P53共表达协同抑制肝细胞癌细胞的增殖、迁移和侵袭[J]. 解剖学报, 2021, 52(2): 251-257.

CHEN Xiao-long WANG Ya-feng HUANG Ping. Co-expression of plasmid-based protein kinase B1-specific small interfering RNA and P53 synergistically inhibits proliferation, migration and invasion of hepatocellular carcinoma cells[J]. Acta Anatomica Sinica, 2021, 52(2): 251-257.

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