苦参碱联合Ⅹ线对肝癌细胞协同杀伤作用的实验

Synergistic Killing Effect and Mechanism of Matrine Combined with X-ray on HepG2 Cells

目的探讨苦参碱和X线对人肝癌Hep G2细胞协同杀伤作用以及诱导细胞凋亡的机制。方法采用MTT法检测细胞活力;ELISA试剂盒检测细胞凋亡;Real-time RT-PCR检测Bcl-2及Bax表达;Western blot检测caspase-9、pro-caspase-9及细胞色素C(cytochondrial-C)的表达;Nobi Flow GOT-IFCC和Nobi Flow LDH-L试剂盒测定LDH、AST水平。结果 0.3 mg/ml苦参碱预处理12 h能显著增加X线(3Gy)对Hep G2细胞的杀伤作用,增加Hep G2细胞凋亡率,抑制Bcl-2的表达,提高胞质内细胞色素C及caspapase-9水平;联合应用X线及苦参碱后人肝细胞中LDH及AST水平、细胞形态均未发生显著改变。结论苦参碱联合X线能协同抑制Hep G2细胞生长、诱导细胞凋亡,同时不会加重对人肝细胞的损害。

Objective To investigate the synergistic killing effect and mechanism of matrine combined with X-ray on human hepatic cancer cells Hep G2. Methods MTT and ELISA were used to detect cell viability and apoptosis, respectively. Real-time RT-PCR was used to examine Bcl-2 and Bax expression. Western blot was used to determine the expression of caspase-9, pro-caspase-9 and cytochondrial-C. Nobi Flow GOT-IFCC and Nobi Flow LDH- L were used to measure the levels of LDH and AST. Results Pretreatment of matrine(3 mg/ml) for 12 h significantly increased the killing effect of X-ray on Hep G2 cells, induced cells apoptosis, inhibited the expression of Bcl-2, and enhanced the levels of cytochondrial-C and caspapase-9 in the cytoplasm. However, the combination of matrine and X-ray did not alter cell morphology, LDH, or AST expression in human hepatocytes. Conclusion The combination treatment of matrine and X-ray could synergistically inhibit the growth of Hep G2 cells by inducing cell apoptosis, without enhancing the damage to human hepatocytes.