大豆苷元及其苯磺酸酯衍生物对肺癌细胞增殖和凋亡的影响

Effects of Daidzein and Its Phenylsulfonate Derivatives on Proliferation and Apoptosis of Lung Cancer Cells

为研究大豆苷元及其3种苯磺酸酯衍生物对非小细胞肺癌细胞的增殖和凋亡的影响,利用CCK-8检测大豆苷元及衍生物处理A549、H1299和HELF细胞后细胞活性的时间-效应关系和剂量-效应关系,并通过Tunel流式细胞术检测衍生物B处理的细胞凋亡情况,RT-qPCR分析TP53和Caspase9 mRNA表达。结果表明:在1.0×10^-5~1.0×10^3μmol·L^-1浓度范围时,加入大豆苷元的浓度每增加10倍,肺正常HELF细胞的活性增加16.11%[95%CI 13.74,18.49](P<0.001),肺癌A549与H1299细胞活性则分别降低3.26%(P<0.001)、3.33%(P<0.001);加入衍生物B的浓度每增加10倍,HELF细胞的活性增加9.92%(P<0.001),A549和H1299细胞活性降低5.23%(P<0.001)、4.32%(P<0.001)。10μmol·L^-1大豆苷元浓度下,作用时间每延长1 h,HELF细胞活性增加1.87%(P<0.001),A549和H1299细胞活性分别降低1.30%(P<0.001)、1.75%(P<0.001),苯磺酸酯衍生物B对HELF细胞活性增加1.72%(P<0.001),A549和H1299细胞降低1.46%(P<0.001)、2.07%(P=0.001)。10μmol·L^-1的衍生物B处理6 h后,HELF细胞相对数量增加了42.66%(P=0.03),A549和H1299细胞相对数量减少20.67%(P=0.035)、18.33%(P=0.043)。衍生物B促进A549和H1299细胞的凋亡的作用约为对照组的1.15倍(P<0.001)、1.24倍(P=0.001),抑制HELF细胞的凋亡作用为对照组的0.84倍(P=0.005),10μmol·L^-1衍生物B处理时肺癌和肺正常细胞的TP53和Caspase9的mRNA表达比对照组高。研究表明:大豆苷元及其3种苯磺酸酯衍生物能够抑制肺癌细胞的增殖活性的同时,增加肺正常细胞增殖活性,大豆苷元苯磺酸酯衍生物B能促进肺癌细胞凋亡并抑制肺正常细胞凋亡,其机制可能与通过P53通路诱导癌细胞凋亡有关。

In order to study the effects of daidzeint and their three phenysulfonate derivatives on proliferation and apoptosis of non-small cell lung cancer(NSCLC)cells, in vitro experiments were conducted on the treatment of A549, H1299 and HELF cells by daidzein and their three derivatives with 1.0×10-5^-1.0×103 μmol·L^-1 and 0-24 h treatment. CCK-8 was used to detect cell activity and apoptosis was detected by Tunel assay. RT-qPCR was used to verify the speculate that these effects maybe involved in TP53 and Caspase9. The results showed that, for the concentration range of 1×10^-5^-1×10^3 μmol·L^-1, when the concentration of daidzein and derivative B was increased 10 times repectively, the proliferation of lung cancer cells could be inhibited to 3.26%(P<0.001), 5.23%(P<0.001) in A549 cells and 3.33%(P<0.001), 4.32%(P<0.001)in H1299 cells, while they were promoted 16.11% [95% CI 13.74,18.49](P<0.001), 9.92%(P<0.001) in HELF cells. By treatment with 10 μmol·L^-1 of daidzein and derivative B from 0 to 24 h, HELF cells were promoted to 1.87%(P<0.001), 1.72%(P<0.001)for every hour extent, while A549 cells were inhibited to 1.30%(P<0.001), 1.46%(P<0.001) and H1299 cells were inhibited to 1.75%(P<0.001), 2.07%(P=0.001)respectively. The relative counts of HELF cells increased by 42.66%(P=0.03), and the counts of H1299 cells decreased by 18.33%(P=0.043)and A549 cells decreased by 20.67%(P=0.035), when treated with 10 μmol·L^-1 derivative B for 6 h. 10 μmol·L^-1 derivative B could promote A549 and H1299 cells to apoptosis with 1.15 times(P<0.001) and 1.24 times(P=0.001), and protect HELF cells out of apoptosis with 0.84 times(P=0.005). The mRNA expression of TP53 and Caspase9 were up-regulation in three cells. To a conclusion, daidzein and their 3 derivatives can inhibit the proliferation of lung cancer cells, and the effect is related to the concentration and time of treatment. derivative B can promote the apoptosis of lung cancer cells. The proliferation of lung cancer cells may be associated with the induction of apoptosis such as TP53 and Caspase9.