槐耳清膏联合顺铂对人肺癌A549细胞和A549/DDP细胞Bax、Bcl-2及Survivin基因表达的影响

Extract of fungi of Huaier combined with cisplatin effects on the expression of Bax, Bcl-2 and Survivin in human lung cancer cells A549 and A549/DDP

目的研究槐耳清膏与顺铂联合使用对人肺癌细胞A549和A549/DDP Bax、Bcl-2以及Survivin基因表达的影响。方法以体外培养的A549和A549/DDP细胞株为模型,将槐耳清膏进行梯度(将10g槐耳清膏溶解于100mL无血清PRMI-1640培养液中,分别制成0、1.0、3.0、5.0、7.0和9.0mg/mL的槐耳清膏溶液)稀释后与12μg/mL顺铂联合使用作用于A549细胞和A549/DDP细胞,并以无药物刺激作为空白对照组,分别采用逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附法(ELISA)检测Bax、Bcl-2以及Survivin基因在A549细胞和A549/DDP细胞表达情况。结果槐耳清膏与顺铂联合使用下,随着槐耳清膏浓度的升高,A549细胞和A549/DDP细胞Bax基因mRNA表达逐渐升高,Bcl-2、Survivin基因mRNA表达逐渐降低,差异均有统计学意义(F=10.21、4.54、7.12,P<0.01,P=0.01,P<0.01;F=12.43、9.17、7.63,P均<0.01)。槐耳清膏与顺铂联合使用的情况下,随着槐耳清膏浓度的升高,A549细胞和A549/DDP细胞Bax蛋白表达量逐渐升高,Bcl-2、Survivin蛋白表达量逐渐降低,差异均有统计学意义(F=8.54、9.38、10.55,P均<0.01;F=15.71、17.98、14.68,P均<0.01)。结论槐耳清膏与顺铂联合使用能够显著提高人肺癌细胞A549和A549/DDP Bax基因表达,降低Bcl-2、Survivin基因表达。

Objective To demonstrate the effect of fungi of Huaier combined with cisplatin effects on the expression of Bax, Bcl-2 and Survivin in human lung cancer cells A549 and A549/DDP. Methods The A549 and A549/DDP cell lines were cultured in vitro. Dissolved 10g of Huaier in 100mL serum-free PRMI-1640 medium and prepared as 0, 1.0, 3.0, 5.0, 7.0 and 9.0mg/mL solution. The gradient dilution Huaier combined with 12 μg/mL of cisplatin were added into A549 and A549/DDP cells. The cells without untreated drugs were as the blank control. The expression of Bax, Bcl-2 and Survivin genes in A549 and A549/DDP cells were detected by RT-PCR and ELISA. Results The mRNA expression of Bax in A549 and A549/DDP cells increased gradually with the higher concentration of Huaier(F=10.21, 4.54 and 7.12;P<0.01, P=0.01 and P<0.01, respectively), but Bcl-2 and Survivin expression decreased significantly(F=12.43, 9.17 and 7.63;P<0.01, P<0.01 and P<0.01, respectively). At the protein level, the expression of Bax also increased gradually(F=8.54, 9.38 and 10.55;P<0.01, P<0.01 and P<0.01, respectively), and the expression of Bcl-2 and Survivin decreased significantly(F=15.71, 17.98 and 14.68;P<0.01, P<0.01 and P<0.01, respectively). Conclusion The combination of Huaier and cisplatin could increase the expression of Bax, and decrease the expression of Bcl-2 and Survivin significantly.