IL-17A对HepG2细胞凋亡的影响及其机制

Effect and Novel Mechanism of IL-ITA on Apoptosis of HepG2 Cell

【目的】探讨白介素-17A（IL-17A）对HepG2细胞凋亡的影响及作用机制。【方法】培养正常肝细胞（L-02）及肝癌细胞（Huh7、HepG2和PLC／PRF／5），荧光实时定量PCR（qRT-PCR）、Westernblot测定其细胞IL-17A表达。然后取HepG2细胞，予重组IL-17A（rIL-17A）和（或）髓细胞白血病因子-1（Mel-1）siRNA处理，流式细胞术分析细胞凋亡率；qRT-PCR、Westernblot检测Mcl-1表达。【结果】相对于L-02细胞，Huh7、HePG2和PLC／PRF/5细胞IL-17AmRNA与蛋白水平明显增加，以HepG2细胞最高（P〈0．01）。rIL-17A使HepG2细胞凋亡减少（Pd0．01），MCl-1表达上调（P〈0．01）。MCl-1siRNA预处理则逆转rIL广17A对HepG2细胞凋亡的抑制作用（Pd0．05）。【结论】肝癌细胞IL-17A呈高表达，IL-17A通过上调MCl-1表达发挥抗HepG2细胞凋亡作用。

[Objective]To explore the effects of interleukin-17A （IL-17A） on apoptosis of HepG2 cell and investigate the novel mechanism. [Methods]Quantitative real-time PCR （qRT-PCR） and Western blot were used to detect IL-17A expression in normal hepatocytes （L-02） and hepatic cancer cell lines （HuhT, HepG2 and PLC/PRF/5）. Subsequently, HepG2 ceils were selected and treated with recombinant IL-17A （rlL-17A） and/or myeloid cell leukemia-1 （Mcl-1） siRNA. Apoptotic rate was analyzed by flow cytometry. The expres- sion of Mcl-1 was measured by qRT-PCR and Western Blot. [Results] Compared to those in L-02 cells, the mRNA and protein levels of IL-17A were significantly increased in Huh7, HepG2 and PLC/PRF/5 cells, and HepG2 cells had the highest expression （ P 〈0.01）. Administration of rIL-17A reduced apoptotic rate and in- creased Mcl-1 expression in HepG2 cells （P 〈0.01）. However, siRNA-mediated knockdown of Mcl-1 re- versed the inhibitory effect of rlL-17A on HepG2 cell apoptosis （ P 〈0.05）.[Conclusion]IL-17A is highly ex- pressed in hepatic cancer cells. IL-17A reduces HepG2 cell apoptosis by upregulating Mcl-1 expression.