CA3诱导肝癌HepG2细胞凋亡及其机制

张舒文; 李晓飞; 于洪丹; 郭莲怡

doi:10.3971/j.issn.1000-8578.2020.19.0948

CA3诱导肝癌HepG2细胞凋亡及其机制

CA3 Induces Apoptosis of Hepatocellular Carcinoma HepG2 Cells and Related Mechanism

摘要

摘要:
目的探讨CA3（CIL56）对肝癌HepG2细胞增殖及凋亡的影响及其机制。
方法体外培养HepG2细胞，分别加入不同浓度CA3（0、0.25、0.5、1.0、2.0和4.0 mmol/L）作用24和48 h。增强型CCK-8试剂检测不同浓度CA3对细胞增殖的影响；流式细胞术检测细胞凋亡率；检测细胞内活性氧（ROS）的变化；Western blot法分析YAP1、Bcl-2、Bax、Caspase-3蛋白的表达。
结果增强型CCK-8试剂检测结果显示，0.25、0.5、1.0、2.0、4.0 mmol/L的CA3作用HepG2细胞24和48 h后，细胞增殖率分别为（80.5±0.3）%、（79.4±0.2）%、（76.2±0.2）%、（76.4±0.1）%、（49.3±0.4）%和（75.3±0.2）%、（64.8±0.3）%、（48.4±0.2）%、（32.2±0.4）%、（31.9±0.2）%。流式细胞术结果显示，CA3浓度升高至2 mmol/L时，凋亡率增加到58.48%。CA3处理后，HepG2细胞内活性氧产量增加，YAP1、Bcl-2蛋白表达降低，Bax、Caspase-3表达增高。
结论 CA3可抑制HepG2细胞增殖并诱导凋亡，其机制可能与细胞内活性氧产量增加，调节YAP1、Bcl-2、Bax及Caspase-3蛋白的表达有关。

Abstract:
Objective To investigate the effect of CA3(CIL56) on the proliferation and apoptosis of hepatocellular carcinoma HepG2 cells and its mechanism.
Methods HepG2 cells were cultured in vitro and treated with different concentrations of CA3 (0, 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L) for 24 and 48 hours, respectively. Enhanced CCK-8 reagent was used to detect the effect of CA3 on cell proliferation. Flow cytometry was utilized to detect cell apoptosis rate. We detected the changes of intracellular reactive oxygen species (ROS). Western blot was utilized to analyze the expression of YAP1, Bcl-2, Bax and Caspase-3 protein.
Results The enhanced CCK-8 reagent test results showed that after 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L CA3 treatment for 24 and 48 hours, the cell proliferation rates were (80.5±0.3)%, (79.4±0.2)%, (76.2±0.2)%, (76.4±0.1)%, (49.3±0.4)% and (75.3±0.2)%, (64.8±0.3)%, (48.4±0.2)%, (32.2±0.4)%, (31.9±0.2)%, respectively. Flow cytometry results showed that when CA3 concentration was increased to 2 mmol/L, the apoptosis rate was increased to 58.48%. CA3 could significantly increase the content of reactive oxygen species in HepG2 cells, reduce the expression of YAP1 and Bcl-2 protein, and increase the expression of Bax and Caspase-3.
Conclusion CA3 could inhibit the proliferation and induce the apoptosis of HepG2 cells. The mechanism may be related to the increase of intracellular reactive oxygen species production and the regulation of YAP1, Bcl-2, Bax and Caspase-3 protein expression.

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