Overexpression of protein phosphatase 2 regulatory subunit B''α gene effect on proliferation and invasion of hepatoma cells
10.3760/cma.j.issn.1007-3418.2019.11.010
- VernacularTitle: 过表达蛋白磷酸酶的调节亚基B''α基因对肝癌细胞增殖和侵袭的影响
- Author:
Huijuan CHEN
1
,
2
;
Peixiao WANG
3
;
Lihong HUANG
2
;
Hongying ZHANG
2
;
Xinguo CHEN
2
;
Qing ZHANG
1
,
2
Author Information
1. Clinical College of General Hospital of Chinese People's Armed Police Force, Anhui Medical University, Beijing 100039, China
2. the Third Medical Centre, Chinese PLA (People’s Liberation Army) General Hospital, Beijing 100039, China
3. Henan Children’s Hospital, Zhengzhou 450018, China
- Publication Type:Journal Article
- Keywords:
Carcinoma, hepatocellular;
Cell proliferation;
Apoptosis;
PPP2R3A gene
- From:
Chinese Journal of Hepatology
2019;27(11):872-878
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the overexpression of protein phosphatase 2 regulatory subunit B''α gene effects on the proliferation and invasion of hepatoma cells.
Methods:Immunohistochemistry method was used to analyze the expression of PPP2R3A in cancerous and paracancerous tissues. Hepatocellular carcinoma cell lines (Huh-7 and HepG2) with stably overexpressing PPP2R3A were constructed by lentiviral vector. Biological behavioral transition in hepatocellular carcinoma cell proliferation, cell cycle, apoptosis, invasion and metastasis were detected by cell counting kit-8 assay (CCK-8), flow cytometry, and transwell assay. A subcutaneous nude tumor mice model was constructed to validate the growth of hepatoma cells. Two independent sample t-tests were used to compare the groups.
Results:The expression of PPP2R3A gene in human hepatocarcinoma tissues was higher than paracancerous tissues. The absorbance (A value) of hepatoma cells was increased (P < 0.05) after overexpression of PPP2R3A gene. The transition from G1-to-S phase was significantly increased i.e., the G1 phase of the cell cycle was reduced (Huh-7: t = 3.04, P = 0.0384; HepG2: t = 4.06, P = 0.0153), while the S phase was increased (Huh-7: t = 3.47, P = 0.0255; HepG2: t = 4.46, P = 0.0112). Early apoptotic rate was decreased (Huh-7: t = 7.34, P = 0.0018; HepG2: t = 4.06, P = 0.0153). The number of Huh-7 cells migrating to the lower chamber was increased (t = 3.18, P = 0.0334), and after the use of matrigel the number of cells reaching to the lower chamber was also increased (t = 2.84, P = 0.0464). The results of animal experiments showed that the subcutaneous tumor growth (t = 4.31, P = 0.0035) was significantly overexpressed in nude mice group. The results of Western blot showed that the expression of PARP and P53 protein in the spliced forms decreased, while the accumulation of β-catenin protein in the liver cancer cells was increased.
Conclusion:Overexpressed PPP2R3A gene may promote proliferation, migration and invasion ability, inhibit apoptosis, induce G1/S phase transition, and participate in the biological behavior of hepatoma cells.