摘要
目的探索二苯乙烯苷(THSG)是否通过调控miR-375表达,进而影响结直肠癌细胞的恶性生物学行为。方法将HT-29细胞分为空白组、实验组、阴性转染组、阳性转染组。空白组和实验组不进行转染,阴性转染组和阳性转染组分别转染NC-inhibitor和miR-375-inhibitor。细胞转染48 h后,空白组给予正常培养,其他3组用含有50 mmol·L^(-1)THSG的培养基培养48 h。用噻唑蓝法测定细胞的增殖情况,用流式细胞仪测定细胞的凋亡情况,用Transwell实验法测定细胞的迁移和侵袭能力,用实时荧光定量聚合酶链反应法检测miR-375的表达水平,用蛋白质印迹法检测细胞中磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)和核因子-κB(NF-κB)p65的磷酸化水平。结果空白组、实验组、阴性转染组和阳性转染组的细胞活力分别为(100.00±3.82)%、(53.88±4.34)%、(55.69±6.03)%和(81.76±6.62)%,凋亡率分别为(6.02±0.44)%、(30.77±2.64)%、(29.70±2.24)%和(14.67±1.37)%,迁移细胞数量分别为(100.00±5.35)、(42.23±3.92)、(41.47±3.43)和(67.67±8.36)个,侵袭细胞数量分别为(78.79±3.45)、(25.73±1.63)、(24.39±1.38)和(44.75±1.66)个,miR-375表达水平分别为1.00±0.07、5.24±0.20、5.28±0.28和1.11±0.08,p-PI3K蛋白相对表达水平分别为1.00±0.04、0.31±0.02、0.31±0.02和0.59±0.05,p-AKT蛋白相对表达水平分别为1.00±0.03、0.31±0.02、0.29±0.02和0.60±0.04,p-NF-κB p65蛋白相对表达水平分别为1.00±0.04、0.10±0.02、0.11±0.01和0.41±0.06。实验组的上述指标与空白组比较,阳性转染组的上述指标与阴性转染组比较,差异均有统计学意义(均P<0.01)。结论THSG可有效抑制结直肠癌细胞的恶性生物学行为,其机制与miR-375介导的PI3K/AKT/NF-κB通路有关。
Objective To reveal whether 2,3,5,4'-Tetrahydroxystilbene 2-O-β-D-glucoside(THSG)affects the malignant biological behavior of colorectal cancer cells through miR-375.Methods HT-29 cells were divided into blank group,experimental group,negative transfection group and positive transfection group.Cells in blank group and experimental group were not transfected,while cells in negative transfection group and positive transfection group were transfected with NC-inhibitor and miR-375-inhibitor,respectively.48 h after transfection,the cells in blank group were cultured normally,while the cells in other groups were cultured with medium containing 50 mmol·L^(-1) THSG for 48 h.HT-29 cell proliferation was determined by methyl thiazolyl tetrazolium.Apoptosis of HT-29 cells was determined by flow cytometry.HT-29 cell migration and invasion were measured by Transwell.The mRNA level of miR-375 was detected by real-time fluorescence quantitative polymerase chain reaction.Phosphorylation levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and nuclear factor-κB(NF-κB)p65 were determined by Western blot method.Results The relative cell vitalities of blank,experimental,negative transfection and positive transfection groups were(100.00±3.82)%,(53.88±4.34)%,(55.69±6.03)%and(81.76±6.62)%;the apoptosis rates were(6.02±0.44)%,(30.77±2.64)%,(29.70±2.24)%and(14.67±1.37)%;the number of migrated cells was 100.00±5.35,42.23±3.92,41.47±3.43 and 67.67±8.36;the number of invasive cells was 78.79±3.45,25.73±1.63,24.39±1.38 and 44.75±1.66;the miR-375 levels were 1.00±0.07,5.24±0.20,5.28±0.28 and 1.11±0.08;the expression levels of p-PI3K protein were 1.00±0.04,0.31±0.02,0.31±0.02 and 0.59±0.05;the protein expression levels of p-Akt were 1.00±0.03,0.31±0.02,0.29±0.02 and 0.60±0.04;the protein expression levels of p-NF-κB p65 were 1.00±0.04,0.10±0.02,0.11±0.01 and 0.41±0.06,respectively.There were significant differences in the above-mentioned indexes between the experimental group and the blank gro
作者
杜宇
苗庄
郭路生
DU Yu;MIAO Zhuang;GUO Lu-sheng(Department of Laboratory,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China;Department of Laboratory,Affiliated Hospital of Jilin Medical College,Jilin 132013,Jilin Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2023年第15期2212-2216,共5页
The Chinese Journal of Clinical Pharmacology
基金
吉林省教育厅“十三五”科学技术基金资助项目(JJKH20170417KJ)。
