拉帕替尼对人皮肤鳞状细胞癌A431细胞生长和凋亡的影响

Effect of lapatinib on the growth and apoptosis of human skin squamous cell carcinoma A431 cells

目的探讨拉帕替尼对人皮肤鳞状细胞癌A431细胞增殖、凋亡以及磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物西罗莫司靶蛋白(PI3K/AKT/mTOR)和无翅型MMTV整合位点家族成员/β联蛋白(Wnt/β-cadherin)信号通路的影响。方法将A431细胞分别置于含有0.1,1.0,10.0,20.0和50.0μmol·L^(-1)拉帕替尼的培养液中培养24和48 h后,用噻唑蓝(MTT)法检测细胞存活率。将A431细胞分为对照组(DMEM培养基培养)和低、中、高浓度实验组(分别用含0.1,1.0和5.0μmol·L^(-1)拉帕替尼的DMEM培养基培养)。用Western blot法检测细胞PI3K/AKT/mTOR及Wnt/β-cadherin信号通路相关蛋白的表达水平。结果在0.1,1.0,10.0,20.0和50.0μmol·L^(-1)拉帕替尼干预24和48 h后均能明显抑制A431细胞的生长,且24 h与48 h的A431细胞存活率均无显著性差异。中、高浓度实验组和对照组的PI3K蛋白表达量分别为0.78±0.06,0.80±0.05和0.74±0.05,p-AKT蛋白表达量分别为0.52±0.04,0.44±0.04和0.75±0.07,p-mTOR蛋白表达量分别为0.35±0.04,0.26±0.02和0.93±0.03,Wnt蛋白表达量分别为0.16±0.01,0.12±0.01和0.95±0.08,β-cadherin蛋白表达量分别为0.25±0.02,0.18±0.01和0.34±0.02,中、高浓度实验组的上述指标与对照组比较,差异均有统计学意义(均P<0.05)。结论拉帕替尼可以抑制A431细胞生长,促进其凋亡及自噬,其机制可能通过阻断PI3K/AKT/mTOR及Wnt/β-cadherin信号通路来实现。

Objective To investigate the effects of lapatinib on proliferation, apoptosis and phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin(PI3 K/AKT/mTOR) and recombinant wingless type MMTV integration site family(Wnt)/β-cadherin signaling pathways of human skin squamous cell carcinoma A431 cells. Methods A431 cells were divided into control group(DMEM culture medium) and the experimental-L,-M,-H groups(DMEM medium containing 0.1, 1.0 and 5.0 μ mol · L^(-1) lapatinib). The cell viability was detected by methyl thiazolyl tetrazolium(MTT). The expression of PI3 K/AKT/mTOR and Wnt/β-cadherin signaling pathway-related proteins was detected by Western blot. Results Lapatinib(0.1, 1.0, 10.0, 20.0 and 50.0 μmol·L^(-1)) significantly inhibited the growth of A431 cells after 24 and 48 h, and there were no significant differences in the survival rate of A431 cells between 24 h and 48 h( all P > 0. 05). The expression levels of PI3 K protein in experimental-M,-H groups and control groups were 0. 78 ± 0. 06,0. 80 ± 0. 05 and 0. 74 ± 0. 05,respectively. The expression of p-Akt protein was 0. 52 ± 0. 04,0. 44 ± 0. 04 and 0. 75 ± 0. 07,respectively. The expression of p-m TOR protein was0. 35 ± 0. 04,0. 26 ± 0. 02 and 0. 93 ± 0. 03, respectively. The expression of Wnt protein was 0. 16 ± 0. 01,0. 12 ± 0. 01,0. 95 ± 0. 08,respectively. The expression of β-cadherin protein was 0. 25 ± 0. 02,0. 18 ± 0. 01,0. 34 ± 0. 02,respectively. The above indexes in the experimental-M,-H groups were significantly higher than those in control group( P < 0. 05). Conclusion Lapatinib can inhibit the growth of A431 cells and promote apoptosis and autophagy,which may be achieved by blocking PI3 K/AKT/m TOR and Wnt/β-cadherin signaling pathways.