紫花前胡苷经通过PI3K/AKT通路对结直肠癌细胞生物学行为的影响

岳峰; 江旗琳; 陈淼; 徐媛媛

doi:10.16190/j.cnki.45-1211/r.2023.01.018

紫花前胡苷经通过PI3K/AKT通路对结直肠癌细胞生物学行为的影响

Influence of nodakenin on biological behavior of colorectal cancer cells through PI3K/AKT pathway

摘要

摘要: 目的:探究紫花前胡苷（NK）经磷脂酰肌醇3激酶（PI3K）/丝氨酸/苏氨酸激酶（AKT）通路对结直肠癌（CRC）细胞生物学行为的影响。方法:将SW480细胞分为SW480组（SW480细胞未做任何处理）、NK低剂量组（L-NK组，40 μmol/L NK）、NK中剂量组（M-NK组，60 μmol/L NK）、NK 高剂量组（H-NK组，80 μmol/L NK）、H-NK+740Y-P 组（80 μmol/L NK+10 μmol/L PI3K激活剂740Y-P处理SW480细胞）。平板克隆检测SW480细胞克隆能力；3-（4, 5-二甲基噻唑-2）2, 5-二苯基四氮唑溴盐（MTT）法检测细胞增殖能力；流式细胞术和Hoechst 33258染色检测SW480细胞凋亡能力；细胞侵袭实验（Transwell）检测SW480细胞侵袭能力；MTT 法检测SW480 细胞黏附情况；蛋白质印迹法（Western blotting）检测黏附、凋亡以及通路相关蛋白水平。结果:与SW480组相比，L-NK组、M-NK组、H-NK组细胞核出现缩小、破碎现象，克隆数、B淋巴细胞瘤-2（Bcl-2）、N-钙黏蛋白（N-cadherin）、Twist相关蛋白1（Twist1）蛋白表达、p-PI3K/PI3K、p-AKT/AKT比例依次显著降低（P＜0.05），细胞生长抑制率、细胞凋亡率以及Bcl-2关联X的蛋白（Bax）、活化的半胱氨酸蛋白酶-3（cleaved-Caspase-3）、E-钙黏附蛋白（E-cadherin）蛋白表达以及细胞黏附抑制率依次显著升高（P＜0.05）；与H-NK 组相比，H-NK+740Y-P 组克隆数、Bcl-2、N-cadherin、Twist1 蛋白表达、p-PI3K/PI3K、p-AKT/AKT 比例显著升高（P＜0.05），细胞生长抑制率、细胞凋亡率以及Bax、cleaved-Caspase-3、E-cadherin蛋白表达以及细胞黏附抑制率显著降低（P＜0.05）。结论:NK可能通过下调PI3K/AKT通路抑制SW480细胞增殖和侵袭，促进其凋亡。

Abstract: Objective: To investigate the influence of nodakenin (NK) on the biological behavior of colorectal cancer (CRC) cells through the phosphatidylinositol 3 kinase (PI3K) /serine/threonine kinase (AKT) pathway.Methods: SW480 cells were divided into SW480 group (SW480 cells were not treated), NK low-dose group (LNK group, 40 μmol/L NK), NK medium-dose group (M-NK group, 60 μmol/L NK), NK high-dose group (H-NK group, 80 μmol/L NK), and H-NK+740Y-P group (SW480 cells treated with 80 μmol/L NK+10 μmol/L PI3K activator 740Y-P).Plate clones were applied to test the cloning ability of SW480 cells.3- (4, 5-dimethylthiazole-2) 2, 5-diphenyltetrazolium bromide (MTT) method was used to detect the proliferation of cells.The apoptosis of SW480 cells was detected by flow cytometry and Hoechst 33258 staining.The invasiveness of SW480 cells was detected by cell invasion test (Transwell).The adhesion of SW480 cells was detected by MTT method.Western blotting was used to detect the levels of adhesion, apoptosis and pathway-related proteins.Results: Compared with SW480 group, the nuclei of L-NK group, M-NK group and H-NK group were shrunk and fragmented, and the number of clones, B-lymphoblastoma-2 (Bcl-2), N-cadherin, Twist related protein 1 (Twist1) protein expression, p-PI3K/PI3K, and p-AKT/AKT ratio were significantly reduced in turn (P＜ 0.05); the cell growth inhibition rate, cell apoptosis rate, and Bcl-2 associated X protein (Bax), activated cysteine proteinase-3 (cleaved Casase-3), E-cadherin expression and the inhibition rate of cell adhesion significantly increased in turn (P＜ 0.05).Compared with the H-NK group, the clone number, Bcl-2, N-cadherin, Twist1 protein expression, p-PI3K/PI3K and p-AKT/AKT ratio in the H-NK+740Y-P group significantly increased (P＜ 0.05).The cell growth inhibition rate, cell apoptosis rate, Bax, cleaved caspase-3, E-cadherin protein expression and cell adhesion inhibition rate significantly decreased (P＜ 0.05).Conclusion: NK may inhibit the proliferation and invasion of SW480 cells and promote their apoptosis by down-regulating PI3K/AKT pathway.

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