刺梨提取物对皮肤鳞状细胞癌A431细胞的抑制作用及对JAK2/STAT3信号通路的影响

Inhibitory effect of prickly pear extract on skin squamous cell carcinoma A431 cells and its effect on JAK2/STAT3 signaling pathway

目的 探讨刺梨提取物对皮肤鳞状细胞癌A431细胞的抑制作用及对JAK2/STAT3信号通路的影响。方法 设A431细胞组,刺梨提取物低、中和高剂量组(0.4、0.8和1.6 mmol/L),各组每孔设6个重复孔,培养72 h。培养结束后,MTT法、苏木素-伊红染色测定细胞增殖、单克隆形成数目水平,划痕实验测定细胞侵袭迁移水平,流式细胞仪及Elisa法测定细胞凋亡水平及凋亡相关蛋白Bcl-2和pro Caspase-3,RT-PCR法、蛋白印迹法测定细胞JAK2和STAT3水平。结果 与对照组比较,刺梨提取物低、中、高剂量组A值、存活率、克隆细胞形成数目、侵袭距离、Bcl-2蛋白表达水平、Jak2、Stat3 mRNA蛋白表达水平降低(P<0.05);随着刺梨提取物剂量的增加,各组细胞A值、存活率、克隆细胞形成数目、侵袭距离、Bcl-2蛋白表达水平、Jak2、Stat3 mRNA蛋白表达水平逐渐降低,剂量-效应关系明显(P<0.05)。与对照组比较,刺梨提取物低、中、高剂量组凋亡率和pro Caspase-3蛋白表达水平升高(P<0.05);随着刺梨提取物剂量的增加,各组细胞凋亡率和pro Caspase-3蛋白表达水平升高,剂量-效应关系明显(P<0.05)。结论 刺梨提取物抑制皮肤鳞状细胞癌A431细胞增殖、侵袭,促进其凋亡;其机制可能与刺梨提取物抑制JAK/STAT信号通路激活有关。

Objective To investigate the inhibitory effect of prickly pear extract on skin squamous cell carcinoma A431 cells and its effect on JAK2/STAT3 signaling pathway. Methods The A431 cell group and the low, medium and high dose groups(0.4, 0.8, 1.6 mmol/L) of Prickly Pear extract were set up. Each well was set up with 6 replicate wells and cultured for 72 h. After the culture, MTT method and hematoxylin staining were used to determine the level of cell proliferation and the number of monoclonal formation. The scratch test was used to determine the level of cell invasion and migration Bcl-2, pro caspase-3. RT-PCR and Western blot were used to measure the levels of JAK2 and STAT3 in cells. Results Compared with the control group, the A value, survival rate, number of clonal cells, invasion distance, Bcl-2 protein expression level, Jak2, Stat3 mRNA protein expression levels in the low, medium and high dose groups of Prickly Pear extract decreased(P<0.05).;With the increase of the dose of prickly pear extract, the A value, survival rate, number of clonal cells, invasion distance, Bcl-2 protein expression level, Jak2, Stat3 mRNA protein expression level of each group gradually decreased, and the dose-effect relationship was obvious(P<0.05). Compared with the control group, the apoptosis rate and the expression level of procaspase-3 protein in the low, medium and high dose groups of prickly pear extract increased(P<0.05). Gradually, the expression level of procaspase-3 protein increased, and the dose-response relationship was obvious(P<0.05). Conclusion Prickly pear extract inhibits the proliferation and invasion of skin squamous cell carcinoma A431 cells and promotes their apoptosis;the mechanism may be related to the inhibition of JAK/STAT signaling pathway activation by Prickly Pear extract.