Cyclin G1对肝癌细胞HepG2放射敏感度的影响及其机制

许刚; 王燕; 吴立广; 薛春泉; 王承伟; 吴朝阳

doi:10.3971/j.issn.1000-8578.2017.17.0039

Cyclin G1对肝癌细胞HepG2放射敏感度的影响及其机制

Effect of Cyclin G1 on Radiosensitivity of Hepatocellular Carcinoma HepG2 Cells and Its Mechanism

摘要

摘要:
目的探讨Cyclin G1对肝癌HepG2细胞放射敏感度的影响及其可能的分子机制。
方法通过转染Cyclin G1-siRNA构建HepG2-Cyclin G1-siRNA细胞，通过成克隆分析观察Cyclin G1对肝癌细胞放射敏感度的影响，流式细胞仪检测肝癌细胞细胞周期分布，ELISA法检测乏氧因子HIF-1α及ROS的表达，Western blot检测凋亡相关蛋白Bcl-2、Bax的表达。
结果 q-PCR显示HepG2-Cyclin G1-siRNA细胞中，Cyclin G1表达下降至正常细胞的43.5%左右，成克隆分析显示Cyclin G1-siRNA可使HepG2细胞放射敏感度增高，与正常对照组相比，放射增敏比SER_Dq为1.41。流式细胞分析显示HepG2-Cyclin G1-siRNA细胞与正常HepG2细胞相比，G₂/M期细胞比例增加，G₀/G₁期细胞比例下降；ELISA显示X线照射可引起HepG2细胞的HIF-1α升高，而Cyclin G1-siRNA可以引起HIF-1α的含量显著下降。ROS含量在X射线照射后表现为轻度升高，Cyclin G1-siRNA可以使ROS含量增加，联合X线照射变化更明显。Western blot显示X线照射及Cyclin G1-siRNA均可使BCL-2表达下降及Bax表达增加，其中HepG2联合X线照射组改变最明显。
结论 Cyclin G1-siRNA可以上调肝癌细胞HepG2的放射敏感度，其机制可能与调节HIF-1α、ROS及凋亡相关蛋白有关。

Abstract:
Objective To investigate the effect of Cyclin G1 on the radiosensitivity of hepatocellular carcinoma HepG2 cells and its possible mechanism.
Methods HepG2-Cyclin G1-siRNA cells was constructed through the transfection of Cyclin G1-siRNA to HepG2 cells, and the effect of Cyclin G1 on radiosensitivity of HepG2 cells was observed. Flow cytometry was used to monitor the change of cell cycle. The expression of hypoxia-inducible factor-1α(HIF-1α) and ROS were detected by ELISA analysis, and Western blot was used to detect the expression of Bcl-2 and Bax.
Results q-PCR showed the expression of Cyclin G1 in HepG2-Cyclin G1-siRNA cells was decreased to 43.5% of normal control group. Clone formation analysis showed that Cyclin G1-siRNA could increase the radiosensitivity of HepG2 cells, SER_Dq=1.41, compared with normal control group. Flow cytometry analysis showed that the cells in G₂/M phase were increased and the cells in G₀/G₁ phase were decreased in HepG2-Cyclin G1-siRNA cells. ELISA analysis showed that the HIF-1α of HepG2 cells could be induced by X-ray irradiation, while Cyclin G1-siRNA could cause a significant decrease in the content of HIF-1α. ROS content in X-ray-irradiated cells showed mild increase, with no significant difference. Cyclin G1-siRNA could increase the content of ROS, moreover, the change was more significant when it combined with X-ray radiation. Western blot showed that X-ray irradiation and Cyclin G1-siRNA could decrease the expression of BCL-2 and increase the expression of Bax, and the changes of HepG2-Cyclin G1-siRNA combined with X-ray irradiation group were the most obvious.
Conclusion Cyclin G1-siRNA could upregulate the radiosensitivity of hepatocellular carcinoma cell line HepG2, and its mechanism may be related to the regulation of HIF-1, ROS and apoptosis-related proteins expressions.

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