摘要
目的探讨紫草素对人胃癌MGC803细胞增殖、迁移、侵袭和凋亡的影响及机制。方法将对数生长期MGC803细胞随机分为空白对照组、紫草素组、紫草素+胰岛素样生长因子-1(IGF-1)组和紫草素+LY294002组。空白对照组细胞在无药培养基中培养,紫草素组细胞在含终浓度10μmol·L^(-1)紫草素的培养基中培养,紫草素+IGF-1组细胞在含终浓度10μmol·L^(-1)紫草素和终浓度10μmol·L^(-1) IGF-1的培养基中培养,紫草素+LY294002组细胞在含终浓度10μmol·L^(-1)紫草素和终浓度30μmol·L^(-1) LY294002的培养基中培养。培养24 h后,采用细胞计数试剂盒-8法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,划痕实验检测细胞迁移能力,Transwell法检测细胞侵袭能力,Western blot法检测细胞中B细胞淋巴瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、细胞色素C(Cyt C)、激活型caspase-3(cleaved caspase-3)、cleaved caspase-9、磷脂酰肌醇3激酶(PI3K)、磷酸化PI3K(p-PI3K)、蛋白激酶B(PKB)、磷酸化PKB(p-PKB)蛋白表达。结果紫草素组MGC803细胞增殖抑制率和凋亡率显著高于空白对照组(P<0.05);紫草素+IGF-1组MGC803细胞增殖抑制率和凋亡率显著低于紫草素组(P<0.05);紫草素+LY294002组MGC803细胞增殖抑制率和凋亡率均显著高于紫草素组(P<0.05)。紫草素组MGC803细胞划痕愈合率和侵袭细胞数显著低于空白对照组(P<0.05);紫草素+IGF-1组MGC803细胞划痕愈合率和侵袭细胞数显著高于紫草素组(P<0.05);紫草素+LY294002组MGC803细胞划痕愈合率和侵袭细胞数显著低于紫草素组(P<0.05)。紫草素组MGC803细胞中Bcl-2蛋白相对表达量显著低于空白对照组(P<0.05),Bax、Cyt C、cleaved caspase-3、cleaved caspase-9蛋白相对表达量及Bax/Bcl-2比值显著高于空白对照组(P<0.05);紫草素+IGF-1组MGC803细胞中Bcl-2蛋白相对表达量显著高于紫草素组(P<0.05),Bax、Cyt C、cleaved caspase-3、cleaved caspase-9蛋白相对表达量及Bax/Bcl-2比值显著低于紫草素组(P<0.05);紫草素+LY294002组MGC803细胞中Bcl-2蛋白相对表达量显著低于紫草素组(P<0.05),Bax、Cyt C、cleaved caspase-3、cleaved caspase-9蛋白相对表达量及Bax/Bcl-2比值显著高于紫草素组(P<0.05)。紫草素组MGC803细胞中p-PI3K、p-PKB蛋白相对表达量及p-PI3K/PI3K、p-PKB/PKB比值显著低于空白对照组(P<0.05),紫草素组和空白对照组MGC803细胞中PI3K、PKB蛋白相对表达量比较差异无统计学意义(P>0.05);紫草素+IGF-1组MGC803细胞中p-PI3K、p-PKB蛋白相对表达量及p-PI3K/PI3K、p-PKB/PKB比值显著高于紫草素组(P<0.05),紫草素+IGF-1组与紫草素组MGC803细胞中PI3K、PKB蛋白相对表达量比较差异无统计学意义(P>0.05);紫草素+LY294002组MGC803细胞中p-PI3K、p-PKB蛋白相对表达量及p-PI3K/PI3K、p-PKB/PKB比值显著低于紫草素组(P<0.05),紫草素+LY294002组与紫草素组MGC803细胞中PI3K、PKB蛋白相对表达量比较差异无统计学意义(P>0.05)。结论紫草素可抑制人胃癌MGC803细胞增殖、迁移、侵袭并促进其凋亡,其作用机制可能与抑制PI3K/PKB信号通路有关。
Objective To investigate the effect and mechanism of shikonin on the proliferation,migration,invasion and apoptosis of human gastric cancer MGC803 cells.Methods The MGC803 cells in the logarithmic growth phase were randomly divided into the blank control group,shikonin group,shikonin+insulin-like growth factor-1(IGF-1)group,and shikonin+LY294002 group.Cells in the blank control group were cultured in drug-free medium,cells in the shikonin group were cultured in the medium containing shikonin with a final concentration of 10μmol·L^(-1),cells in the shikonin+IGF-1 group were cultured in the medium containing shikonin with a final concentration of 10μmol·L^(-1) and IGF-1 with a final concentration of 10μmol·L^(-1),and cells in the shikonin+LY294002 group were cultured in the medium containing shikonin with a final concentration of 10μmol·L^(-1) and LY294002 with a final concentration of 30μmol·L^(-1).After 24 h of culture,the cell proliferation was detected by cell counting kit-8,the cell apoptosis was detected by flow cytometry,the cell migration was detected by scratch assay,and the cell invasion was detected by Transwell assay.The expression levels of B cell lymphoma-2(Bcl-2),Bcl-2 related X protein(Bax),cytochrome C(Cyt C),cleaved caspase-3,cleaved caspase-9,phosphoinositide 3 kinase(PI3K),phosphorylated PI3K(p-PI3K),protein kinase B(PKB),and phosphorylated PKB(p-PKB)proteins were measured by using Western blot.Results The MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin group were significantly higher than those in the blank control group(P<0.05);the MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin+IGF-1 group were significantly lower than those in the shikonin group(P<0.05);and the MGC803 cell proliferation inhibition rate and apoptosis rate in the shikonin+LY294002 group were significantly higher than those in the shikonin group(P<0.05).The MGC803 cell scratch healing rate and the number of invasive cells in the shikonin group were significantly lower than those in the blank control group(P<0.05);the MGC803 cell scratch healing rate and the number of invasive cells in the shikonin+IGF-1 group were significantly higher than those in the shikonin group(P<0.05);and the MGC803 cell scratch healing rate and the number of invasive cells in the shikonin+LY294002 group were significantly lower than those in the shikonin group(P<0.05).The relative expression level of Bcl-2 protein in MGC803 cells in the shikonin group was significantly lower than that in the blank control group(P<0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly higher than those in the blank control group(P<0.05);the relative expression level of Bcl-2 protein in MGC803 cells in the shikonin+IGF-1 group was significantly higher than that in the shikonin group(P<0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly lower than those in the shikonin group(P<0.05);and the relative expression level of Bcl-2 protein in MGC803 cells in the shikonin+LY294002 group was significantly lower than that in the shikonin group(P<0.05),while the relative expression levels of Bax,Cyt C,cleaved caspase-3 and cleaved caspase-9 proteins and the Bax/Bcl-2 ratio were significantly higher than those in the shikonin group(P<0.05).The relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin group were significantly lower than those in the blank control group(P<0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin group and the blank control group(P>0.05);the relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin+IGF-1 group were significantly higher than those in the shikonin group(P<0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin+IGF-1 group and the shikonin group(P>0.05);and the relative expression levels of p-PI3K and p-PKB proteins and the ratios of p-PI3K/PI3K and p-PKB/PKB in MGC803 cells in the shikonin+LY294002 group were significantly lower than those in the shikonin group(P<0.05),and there was no statistically significant difference in the relative expression levels of PI3K and PKB proteins in MGC803 cells between the shikonin+LY294002 group and the shikonin group(P>0.05).Conclusion Shikonin can inhibit the proliferation,migration and invasion and promote the apoptosis of human gastric cancer MGC803 cells,which may be related to its inhibition of the PI3K/PKB signaling pathway.
作者
张欣
霍浩然
薛佳栋
武星
刘帆
任继中
袁增江
ZHANG Xin;HUO Haoran;XUE Jiadong;WU Xing;LIU Fan;REN Jizhong;YUAN Zengjiang(Department of General Surgery,Handan Central Hospital,Handan 056000,Hebei Province,China)
出处
《新乡医学院学报》
CAS
2024年第6期515-522,528,共9页
Journal of Xinxiang Medical University
基金
河北省邯郸市科学技术研究与发展计划项目(编号:19422083016ZC)。
关键词
紫草素
胃癌
磷脂酰肌醇3激酶/蛋白激酶B通路
细胞增殖
细胞凋亡
shikonin
gastric cancer
phosphoinositide 3 kinase/protein kinase B pathway
cell proliferation
cell apoptosis
