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目的:研究松针油对人肝癌HepG2细胞的作用。方法:水蒸气蒸馏法提取松针油,GC-MS分析其成分,不同浓度松针油处理HepG2细胞后,用MTT法测定HepG2细胞的生长抑制率。HE染色及电镜下观察细胞凋亡形态的改变。流式细胞仪技术检测细胞凋亡率;流式细胞仪检查bcl-2基因表达水平;TRAP法测定端粒酶活性的变化。结果:MTT法证明松针油可显著抑制HepG2细胞生长,抑制率与药物浓度正相关,呈现明显的量-效和时-效关系。流式细胞仪可以检测到细胞凋亡,0.125~8 mg/L的松针油能引起HepG2细胞凋亡,且呈时间依赖性及剂量依赖性。电镜下可观察到核浓缩及核碎裂等典型的细胞凋亡特征。松针油在诱导细胞凋亡的同时伴随端粒酶活性下调和bcl-2表达水平下降。结论:松针油诱导肝癌HepG2细胞凋亡,并抑制端粒酶活性,bcl-2参与了松针油诱导的肝癌HepG2细胞凋亡过程中端粒酶活性的调控。
Objective: To study the effect of pine needles on HepG2 cells. Methods: The pine needle oil was extracted by steam distillation. The composition of HepG2 cells was analyzed by GC-MS. The growth inhibition rates of HepG2 cells were determined by MTT assay after HepG2 cells were treated with different concentrations of pine needle oil. The changes of apoptotic morphology were observed by HE staining and electron microscopy. Flow cytometry was used to detect the apoptosis rate. The expression of bcl-2 gene was detected by flow cytometry. The telomerase activity was measured by TRAP. Results: MTT showed that pineal oil could significantly inhibit the growth of HepG2 cells. The inhibition rate was positively correlated with the drug concentration, showing a significant dose-effect and time-effect relationship. Flow cytometry can detect apoptosis, pineal oil of 0.125 ~ 8 mg / L can cause apoptosis of HepG2 cells in a time-and dose-dependent manner. Electron microscopy can be observed nuclear condensation and nuclear fragmentation and other typical characteristics of apoptosis. Pine needle oil induces apoptosis accompanied by down-regulation of telomerase activity and decrease of bcl-2 expression. CONCLUSION: Pine needle oil induces apoptosis and inhibits telomerase activity in hepatocellular carcinoma HepG2 cells. Bcl-2 is involved in the regulation of telomerase activity during pineal oil-induced apoptosis in HepG2 cells.