乙酰紫草素对耐药结肠癌细胞增殖、侵袭和迁移的影响  

Effects of acetylshikonin on proliferation,invasion and migration of multidrug resistant colon cancer cells

作　　者：李莉 朱雨 姜京植[2] 金雪梅 朴丽花[2] LI Li;ZHU Yu;JIANG Jing-zhi;JIN Xue-mei;PIAO Li-hua(Department of Anatomy,Yanbian University College of Medicine,Yanji 133002,Jilin Province,China;Department of Histology and Embryology,Yanbian University College of Medicine,Yanji 133002,Jilin Province,China;Department of Pathology,Affiliated Hospital of Yanbian University,Yanji 133002,Jilin Province,China)

机构地区：[1]延边大学医学院解剖学教研室,吉林延吉133002 [2]延边大学医学院组织胚胎学教研室,吉林延吉133002 [3]延边大学附属医院病理科,吉林延吉133002

出　　处：《中国临床药理学杂志》2024年第19期2842-2846,共5页The Chinese Journal of Clinical Pharmacology

基　　金：吉林省科技厅基金资助项目(YDZJ202201ZYTS163)。

摘　　要：目的探究乙酰紫草素对奥沙利铂耐药结肠癌HCT116细胞(HCT116/L-OHP)增殖、侵袭迁移的影响。方法将HCT116/L-OHP细胞分为空白组、对照组和低、中、高剂量实验组。对照组用10μmol·L^(-1)奥沙利铂处理细胞;低、中、高剂量实验组分别用1.25、2.50和5.00μmol·L^(-1)乙酰紫草素和10μmol·L^(-1)奥沙利铂共同处理细胞;空白组给予常规培养。用细胞计数试剂盒-8(CCK-8)法检测HCT116/L-OHP细胞增殖情况,用流式细胞仪检测细胞凋亡情况,用Transwell实验法检测细胞迁移和侵袭能力,用蛋白质印迹法检测耐药侵袭相关蛋白P-糖蛋白(P-gp)、基质金属蛋白酶2(MMP2)、核因子κB(NF-κB)和缺氧诱导因子-1α(HIF-1α)蛋白的表达情况。结果空白组、对照组和低、中、高剂量实验组的细胞抑制率分别为0、(8.27±0.01)%、(10.53±0.02)%、(34.17±0.01)%和(48.47±0.05)%,细胞凋亡率分别为(0.13±0.02)%、(1.37±1.04)%、(9.73±0.87)%、(26.71±4.26)%和(40.75±4.70)%,侵袭细胞数分别为(130.70±9.81)、(127.10±9.21)、(71.83±3.57)、(28.83±1.87)和(19.63±6.11)个,迁移细胞数分别为(150.50±10.17)、(148.40±8.13)、(94.58±4.09)、(63.98±5.09)和(31.85±5.50)个,P-gp蛋白相对表达水平分别为0.91±0.01、0.89±0.02、0.75±0.04、0.61±0.07和0.25±0.03,MMP2蛋白相对表达水平分别为1.24±0.01、1.22±0.02、0.96±0.01、0.53±0.01和0.16±0.02,NF-κB-p65蛋白相对表达水平分别为1.12±0.12、1.07±0.01、0.78±0.01、0.64±0.02和0.31±0.03,HIF-1α相对表达水平分别为0.65±0.04、0.52±0.03、0.41±0.02、0.35±0.03和0.09±0.01。低、中、高剂量实验组的上述指标与空白组比较,在统计学上差异均有统计学意义(均P<0.05)。结论乙酰紫草素联合奥沙利铂可显著抑制HCT116/L-OHP细胞的增殖、侵袭和迁移,诱导细胞凋亡,其机制可能与抑制P-gp、MMP2的表达及NF-κB/HIF-1α信号活化有关。Objective To investigate the effects of acetylshikonin on proliferation,invasion and migration of oxaliplatin-resistant human colon cancer HCT116 cells(HCT116/L-OHP).Methods HCT116/LOHP cells were divided into blank group,control group,experimental-L group,experimental-M group and experimental-H group.The control group was treated with 10μmol·L^(-1)oxaliplatin.The experimental-L,experimental-M,experimental-H groups were treated with 1.25,2.50 and 5.00μmol·L^(-1)acetylshikin and 10μmol·L^(-1)oxaliplatin,respectively.The blank group was given routine culture.The changes of HCT116/L-OHP cell proliferation were detected by cell counting kit-8(CCK-8)method;flow cytometry was used to evaluate the apoptosis of cells;Transwell assay was used to detect the changes of cell migration and invasion ability;Western blot was used to detect the expressions of P-glycoprotein(P-gp),matrix metallo-proteinases 2(MMP2),nuclear factor kappa-B(NF-κB)/and hypoxia induced factor-1α(HIF-1α)proteins.Results The cell inhibition rates of the blank group,control group and experimental-L,-M,-H groups were 0,(8.27±0.0)%,(10.53±0.02)%,(34.17±0.01)%and(48.47±0.05)%;cell apoptosis rates were(0.13±0.02)%,(1.37±1.04)%,(9.73±0.87)%,(26.71±4.26)%and(40.75±4.70)%;invading cells were 130.70±9.81,127.10±9.21,71.83±3.57,28.83±1.87 and 19.63±6,11;the number of migration cells was 150.50±10.17,148.40±8,13,94.58±4.09,63.98±5.09 and 31.85±5.50;the relative expression levels of P-gp protein were 0·91±0.01,0.89±0.02,0.75±0.04,0.61±0.07 and 0.25±0.03;the relative expression levels of MMP2 protein were 1.24±0.01,1.22±0.02,0.96±0.01,0.53±0.01 and 0.16±0.02;the relative expression levels of NF-κB-p65 were 1.12±0.12,1.07±0.01,0.78±0.01,0·64±0.02 and 0.31±0.03;the relative expression levels of HIF-1αwere 0.65±0.04,0.52±0.03,0.41±0.02,0.35±0.03 and 0,09±0.01,respectively,The above indicators in the experimental-L,-M,-H groups showed statistically significant differences compared to those of blank group(all P<0.05).Co

关 键 词：乙酰紫草素 结肠癌 P糖蛋白 耐药性 侵袭 转移 

分 类 号：R28[医药卫生—中药学]