汉防己甲素诱导神经母细胞瘤株TGW凋亡作用的实验研究

Study on the efficacy of Tetrandrine in inducing the apoptosis of neuroblastoma cells

目的为评价汉防己甲素（TET）应用于神经母细胞瘤的可能性，应用TET处理人神经母细胞瘤株TGW，观察其作用情况。方法采用MTT比色法观察TET对神经母细胞瘤增生的影响，琼脂凝胶电泳法观察DNA片断化的梯形条带，流式细胞仪观察TET对神经母细胞瘤凋亡水平影响，实时定量PCR法了解Bcl-2家族（Bcl-2，Bcl-XL，Bad，Bax）表达的变化。结果25～0．5μM TET能明显抑制TGW细胞，25μM汉防己甲素作用48h最大抑制率为（95．32±2．46）％；琼脂糖凝胶电泳法观察到25肛MTET处理12、24、36、48h后出现典型的凋亡表现；流式细胞仪Annexin及PI双染发现25pMTET孵育24h肿瘤细胞的凋亡率明显上升；实时定量PCR发现25肛MTET处理24h后Bax基因表达明显增加。结论汉防己甲素能诱导人神经母细胞瘤株TGW凋亡，上调Bax基因的表达。

Objective Tetrandrine （TET）, a bisbenylisoquinoline alkaloid extracted from the dried root of Stephenia tetrandra S Moore, exhibits very broad pharmacological actions, including anti-tumor activity. To explore the potential clinical efficacy of Tetrandrine on treating neuroblastoma, the effect on human neuroblastom cell line TGW was studied. Methods MTT was used to measure the levels of the proliferation of TGW cultured with Tetrandrine in different concentrations. The DNA ladder was detected with agarose gel electrophoresis. Flow cytometry was used to test the apoptotic level of the cell lines treated with Tetrandrine. Real-time PCR was used to detect the expression of Bcl-2 families（Bcl-2, Bcl-XL, BAD, BAX） mRNA after Tetrandrine treatment. Results The growth of TGW cell lines were inhibited by Tetrandrine of 25 - 0.5 μM. The maximum growth inhibition rate of Tetrandrine was （95.32 ± 2.46） %, occured at 48h with 25μM,After the cell line was incubated in 25μM Tetrandrin for 12 to 48 hours, typical apoptosis was observed with agarose gel electrophoresis. Flow cytometry with Annexin V-FITC/PI double labeling showed the apoptotic level of TGW cell lines upgraded when it was incubated in 25 μM Tetrandrine for 24 hours. Bax gene expression was increased when they were incubated in 25 μM Tetrandrine for 24 hours. Conelusions Tetrandrine could promote apoptosis of neuroblastoma cell line TGW and up-regulate the expression of Bax gene.