对香豆酸诱导弥漫性大B细胞淋巴瘤细胞内质网应激介导的凋亡

Induction of endoplasmic reticulum stress-mediated apoptosis in diffuse large B-cell lymphoma cells by p-coumaric acid

目的探讨对香豆酸诱导弥漫性大B细胞淋巴瘤细胞内质网应激介导的凋亡。方法选择弥漫性大B细胞淋巴瘤细胞(OCI-LY1),以不同浓度对香豆酸(0、0.4、0.8、1.6、3.2 mmol/L)作用于OCI-LY1细胞,采用CCK8法检测其抑制率及半数抑制浓度(IC_(50));随后设置分组:(1)空白对照组、对香豆酸(1/2 IC_(50)组、IC_(50)组、2 IC_(50)组);(2)空白对照组、对香豆酸IC_(50)组、4-PBA组(5 mmol/L)、4-PBA+IC_(50)组,采用CCK8检测细胞活性;流式细胞术检测细胞凋亡;蛋白质印迹法(Western blot)检测细胞GRP78、ATF6、CHOP、p-PERK、p-IRE1α蛋白相对表达量。结果对香豆酸以剂量依赖性方式抑制OCI-LY1细胞增殖,其IC_(50)值为2.601 mmol/L;与空白对照组相比,1/2 IC_(50)组、IC_(50)组、2 IC_(50)组细胞抑制率、细胞凋亡及GRP78、ATF6、CHOP、p-PERK、p-IRE1α蛋白表达明显升高(P<0.01);与IC_(50)组相比,4-PBA组、4-PBA+IC_(50)组细胞抑制率、细胞凋亡及GRP78、ATF6、CHOP、p-PERK、p-IRE1α蛋白表达明显降低(P<0.01);与4-PBA组相比,4-PBA+IC_(50)组细胞抑制率、细胞凋亡及GRP78、ATF6、CHOP、p-PERK、p-IRE1α蛋白表达明显升高(P<0.01)。结论对香豆酸抑制OCI-LY1细胞增殖,可能通过激活OCI-LY1细胞内质网应激从而诱导细胞凋亡。

Objective To investigate induction of endoplasmic reticulum stress-mediated apoptosis in diffuse large B-cell lymphoma cells by p-coumaric acid.Methods Diffuse large B-cell lymphoma cell line OCI-LY1 was treated with pcoumaric acid(0,0.4,0.8,1.6 and 3.2 mmol/L)and the inhibition rate and half-inhibition concentration(IC_(50))was determined by CCK8 assays.Subsequently,groupings were set as follows:(1)control group and p-coumaric acid groups(1/2 IC_(50),IC_(50) and 2 IC_(50)).(2)control group,p-coumaric acid IC_(50) group,4-PBA group(5 mmol/L)and 4-PBA+IC_(50) group.Cell activity was assessed by CCK8 assays.Apoptosis was detected by flow cytometry.Relative expression of GRP78,ATF6,CHOP,p-PERK and p-IRE1αproteins was detected by Western blot.Results p-Coumaric acid inhibited OCI-LY1 cell proliferation in a dose-dependent manner with an IC_(50) value of 2.601 mmol/L.Compared with the control group,the cell inhibition rate,apoptosis,and GRP78,ATF6,CHOP,p-PERK and p-IRE1αprotein expression were significantly increased in the 1/2 IC_(50),IC_(50) and 2 IC_(50) groups(P<0.01).Compared with the IC_(50) group,the cell inhibition rate,apoptosis and GRP78,ATF6,CHOP,p-PERK,p-IRE1αprotein expression were significantly decreased in the 4-PBA group and 4-PBA+IC_(50) group(P<0.01).Compared with the 4-PBA group,cell inhibition rate,apoptosis and GRP78,ATF6,CHOP,p-PERK and p-IRE1αprotein expression were significantly increased in the 4-PBA+IC_(50) group(P<0.01).Conclusions p-Coumaric acid inhibits OCI-LY1 cell proliferation,possibly by activating endoplasmic reticulum stress and inducing apoptosis.