苦杏仁苷调控PI3K/Akt/mTOR通路对子宫内膜癌细胞增殖、侵袭及凋亡的影响

Effects of Amygdalin on Proliferation,Invasion and Apoptosis of Endometrial Cancer Cells through Inhibiting PI3K/Akt/mTOR Pathway

目的研究苦杏仁苷对子宫内膜癌细胞RL95-2及HEC-1B增殖、侵袭及凋亡的影响。方法使用8、16、32、64和128mg/L浓度苦杏仁苷处理RL95-2、HEC-1B细胞24、48和72h后,采用CCK-8法检测各组细胞增殖能力并计算苦杏仁苷的半数抑制浓度(IC50);Transwell试验检测苦杏仁苷对细胞侵袭能力的影响;流式细胞仪检测各组细胞凋亡情况;Western blot试验检测凋亡相关基因Bcl-2、Bax蛋白以及PI3K/Akt/mTOR信号通路相关蛋白表达情况。结果苦杏仁苷呈浓度及时间依赖性抑制子宫内膜癌RL95-2、HEC-1B细胞生长活性,48h时半数抑制浓度(IC50)约为40mg/L;苦杏仁苷能抑制RL95-2、HEC-1B细胞穿膜细胞数[(227.67±11.06)个比(459.33±24.75)个,(188.67±11.14)个比(412.33±14)个,P均<0.01];经苦杏仁苷处理后RL95-2、HEC-1B细胞凋亡率升高[(7.85±0.71)%比(1.49±0.08)%、(8.55±0.69)%比(1.61±0.13)%,P均<0.01],凋亡蛋白Bcl-2表达下降[(0.19±0.03)比(0.36±0.04)、(0.25±0.04)比(0.39±0.05),P均<0.01],Bax表达升高[(0.37±0.06)比(0.18±0.04)、(0.41±0.06)比(0.21±0.03),P均<0.01];苦杏仁苷能显著下调RL95-2、HEC-1B细胞的PI3K[(0.42±0.04)比(0.78±0.05)、(0.36±0.05)比(0.63±0.03),P均<0.01]、Akt[(0.73±0.05)比(0.92±0.05)、(0.65±0.03)比(0.86±0.04),P均<0.01]及mTOR[(0.25±0.04)比(0.93±0.06)、(0.34±0.05)比(0.88±0.04),P均<0.01]磷酸化水平,抑制PI3K/Akt/mTOR通路活化。结论苦杏仁苷通过抑制PI3K/Akt/mTOR信号通路活化,促进子宫内膜癌RL95-2、HEC-1B细胞凋亡,降低其增殖活性,抑制细胞侵袭功能,从而发挥抗肿瘤作用。

Objective To explore the effects of amygdalin on the proliferation, invasion and apoptosis of endometrial cancer cells RL95-2 and HEC-1 B. Methods RL95-2 and HEC-1 B cells were treated with amygdalin at concentrations of 8, 16, 32, 64 and 128 mg/L for 24, 48 and 72 h. CCK-8 method was used to detect the cell proliferation and calculate the half inhibitory concentration(IC50) of amygdalin;Transwell test was used to test the effect of amygdalin on cell invasion;flow cytometry was used to check cell apoptosis;Apoptosis-related genes Bcl-2, Bax protein and PI3K/Akt/mTOR signaling pathway related protein expression were detected by Western blot.Results Amygdalin inhibited the growth activity of endometrial cancer RL95-2 and HEC-1B cells in a concentration and time dependent manner, and the inhibitory concentration(IC50) of amygdalin was about 40 mg/L at 48 h.Amygidin inhibited the number of transmembrane cells of RL95-2 and HEC-1B cells [(227.67 ±11.06) vs(459.33±24.75), and(188.67±11.14) vs(412.33±14), P<0.01, respectively]. After treatment with amygidin, the apoptotic rate of RL95-2 and HEC-1 B cells increased [(7.85±0.71)% vs(1.49±0.08)%, and(8.55±0.69)% vs(1.61±0.13)%, P<0.01, respectively], the expression of apoptotic protein Bcl-2 decreased [(0.19±0.03) vs(0.36±0.04), and(0.25±0.04) vs(0.39±0.05), P<0.01, respectively], and Bax expression was increased [(0.37±0.06) vs(0.18±0.04), and(0.41±0.06) vs(0.21±0.03), P<0.01, respectively]. Amygdalin significantly decreased the phosphorylation levels of PI3 K [(0.42 ±0.04) vs(0.78 ±0.05), and(0.36 ±0.05) vs(0.63 ±0.03), P <0.01, respectively],Akt [(0.73±0.05) vs(0.92±0.05), and(0.65±0.03) vs(0.86±0.04), P<0.01, respectively], and mTOR [(0.25±0.04)vs(0.93±0.06), and(0.34±0.05) vs(0.88±0.04), P<0.01, respectively], thus inhibited the activation of PI3 K/Akt/mTOR pathway. Conclusion AAmygdalin can inhibit the activation of PI3K/Akt/mTOR signaling pathway, promote the apoptosis of endometrial cancer RL95-2 and HEC-1 B cells, reduce their proliferation activity and inhibit cell invasion function, thus playing an anti-tumor role.