基于p38-MAPK信号通路探讨和厚朴酚对结直肠癌细胞增殖、凋亡的影响

目的基于p38-丝裂原活化蛋白激酶(p38-mitogen activated protein kinases,p38-MAPK)探讨和厚朴酚(honokiol,HNK)对结直肠癌细胞增殖和凋亡的影响.方法将体外培养HCT116细胞分为对照组(0 μmol/L)、HNK低剂量(20 μmol/L)、中剂量(40 μmol/L)和高剂量组(80μmol/L)。采用CCK-8法和克隆形成试验检测细胞增殖,流式细胞仪检测细胞凋亡,Western印迹检测p38、p-P38、细胞增殖指数67(cell proliferation index 67,Ki67)、B细胞淋巴瘤/白血病-2(B-cell lymphoma-2,Bcl-2)、BCL2相关X蛋内(B-cell lymphoma-2 associated X protein,Bax)和裂解的半胱氨酸天冬氨酸蛋内酶3(cleaved Caspase-3)蛋白表达。将HCT116细胞分为空内对照组(0μmol/L)、HNK组(80μmol/L)、SB203580组(10 μmoL/L)和SB203580+HNK组(SB203580 10μmol/L+HNK 80 μmol/L),检测SB203580对各组细胞增殖、凋亡以及Ki67、cleaved Caspase3蛋白表达的影响。结果与对照组比较,HNK中高剂量组HCT116细胞的相对增殖率和克隆形成率明显降低,细胞凋亡率明显升高,Ki-67、Bcl-2蛋白表达明显下降,p-p38/p38、Bax和cleaved Caspase-3蛋白表达明显升高(P<0.05),且随着HNK剂量增加,其作用明显增加(P<0.05)。与空白对照组比较,SB203580组细胞相对增殖率明显升高,细胞凋亡率明显下降,Ki67蛋内表达明显升高,cleaved Caspase-3蛋白表达明显下降(P<0.05);与SB203580组比较,SB203580+HNK组细胞相对增殖率明显下降,细胞凋亡率明显升高,Ki67蛋白表达明显下降,cleaved Caspase-3蛋白表达明显升高(P<0.05)。结论HNK可以激活p38-MAPK信号通路,抑制HCT116细胞增殖,诱导细胞凋亡。

Effects of Honokiol on Proliferation and Apoptosis of Colorectal Cancer Cells via p38-MAPK Signaling Pathway

Objective To explore the effects of honokiol(HNK)on proliferation and apoptosis of colorectal cancer(CRC)cells based on p38-mitogenactivated protein kinase(p38-MAPK)signaling pathway.Methods HCT116 cells were cultured in vitro and divided into control group(0 μmol/L),low-dose HNK(20μmol/L),medium-dose HNK(40μmol/L)and high-dose HNK groups(80μmol/L).Cell proliferation was detected by CCK-8 and clone formation assay.Apoptosis was detected by flow cytometry.The expression levels of p38,p-p38,cell proliferation index 67(Ki67),B-cell lymphoma-2(Bcl-2),B-cell lymphoma-2 associated X protein(Bax)and cleaved cysteine aspartase 3(cleaved Caspase-3)were detected by Western blotting.HCT116 cells were divided into blank control group(0 μmol/L),HNK group(80μmol/L),SB203580 group(10 μmol/L)and SB203580+HNK group(SB20358010 μmol/L+HNK 80μmol/L).The effects of SB203580 on cells proliferation,apoptosis,expression of Ki67 and Cleaved caspase3 in each group were detected.Results Compared with control group,relative proliferation rate and clone formation rate of HCT116 cells were significantly decreased,apoptosis rate was significantly increased,expression levels of Ki67 and Bcl-2 proteins were significantly decreased,and expression levels of p-p38/p38,Bax and cleaved Caspase-3 proteins were significantly increased in medium-dose and high-dose HNK groups(P<0.05).With the increase of HNK dose,its effects were significantly increased(P<0.05).Compared with blank control group,relative proliferation rate of cells was significantly increased,apoptosis rate was significantly decreased,expression of Ki67 protein was significantly increased,and expression of cleaved Caspase-3 protein was significantly decreased in SB203580 group(P<0.05).Compared with the SB203580 group,relative proliferation rate of cells was significantly decreased,apoptosis rate was significantly increased,expression of Ki67 protein was significantly decreased,and expression of cleaved Caspase-3 protein was significantly increased in SB203580+HNK group(P<0.05).Conclusion HNK can activate p38-MAPK signaling pathway,inhibit proliferation of HCT116 cells and induce apoptosis.