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.
目的： 研究过表达蛋白磷酸酶2A的调节亚基B''α对肝癌细胞增殖与侵袭能力的影响。 方法： 通过免疫组织化学法分析癌组织及癌周组织中PPP2R3A表达；用慢病毒载体构建稳定过表达PPP2R3A的肝癌细胞系(Huh-7和HepG2)；通过CCK-8、流式细胞仪、transwell小室实验分别检测稳转肝癌细胞系增殖、周期、凋亡以及侵袭转移的生物学行为的改变，并构建裸鼠皮下瘤模型验证肝癌细胞生长情况。计量资料组间比较用t检验。 结果： 人肝癌癌组织中PPP2R3A基因的表达高于癌旁；过表达PPP2R3A基因后，肝癌细胞的吸光度值（A值）增加（P < 0.05）；细胞周期G(1)期向S期转换显著增多，即细胞周期G(1)期减少(Huh-7：t = 3.04，P = 0.038 4；HepG2：t = 4.06，P = 0.015 3)，S期增多(Huh-7：t = 3.47，P = 0.025 5；HepG2：t = 4.46，P = 0.011 2)；早期凋亡率减少（Huh-7：t = 7.34，P = 0.001 8；HepG2：t = 4.06，P = 0.015 3）；Huh-7细胞中，迁移到下室的数目明显增多（t = 3.18，P = 0.033 4)，铺上基质胶后到达下室的细胞数目同样增多（t = 2.84，P = 0.046 4)；动物实验结果显示其过表达组的裸鼠皮下瘤生长显著增加（t = 4.31，P = 0.003 5）。蛋白质印迹法实验结果显示剪切体的PARP和P53蛋白表达减少，而β-catenin蛋白在肝癌细胞的积累增多。 结论： 过表达PPP2R3A基因可能促进肝癌细胞的增殖、迁移与侵袭能力，抑制凋亡，诱导G(1)/S期转换，参与影响肝癌细胞生物学行为。.
Keywords: Apoptosis; Carcinoma, hepatocellular; Cell proliferation; PPP2R3A gene.