松花粉对于肝癌细胞株SMMC-7721的生物学作用及与α-烯醇酶及其下游PI3K/AKT信号通路蛋白的关联作用  

Biological effect of pine pollen on hepatocellular carcinoma cell line SMMC-7721 and its association withα-Enolase and the proteins of its downstream PI3K/AKT signaling pathway

作　　者：徐伟鑫 王春芳[1] 罗艳红[1] Xu Weixin;Wang Chunfang;Luo Yanhong(School of Laboratory Medicine,Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)

机构地区：[1]右江民族医学院医学检验学院,广西百色533000

出　　处：《右江民族医学院学报》2023年第1期15-21,共7页Journal of Youjiang Medical University for Nationalities

基　　金：国家自然科学基金项目(81960303);右江民族医学院校级科研课题项目(右医科字〔2021〕38号)。

摘　　要：目的探讨松花粉作用于肝癌细胞株SMMC-7721的增殖和侵袭作用的影响,以及与之相关的磷脂酰肌醇3-激酶/蛋白质丝氨酸苏氨酸激酶(PI3K/AKT)信号通路蛋白的表达变化。方法首先对松花粉水提液中总多糖、总黄酮和锌离子3种主要抗癌成分的浓度进行检测。再用未加松花粉处理的SMMC-7721细胞作为对照组,分别用一系列浓度的松花粉处理12 h、24 h于SMMC-7721细胞,用CCK8法检测细胞增殖及其最优半抑制浓度(IC50),并且以SMMC-7721细胞的IC50作为松花粉处理组的浓度进行后续Transwell法检测细胞侵袭、Western Blot法检测α-烯醇酶(ENO1)及下游PI3K/AKT信号通路蛋白的表达变化。结果CCK8法检测SMMC-7721细胞在松花粉处理24 h后的IC50为16.78μg/μL,与12 h的23.87μg/μL相比有更好的效果;故后续实验采用松花粉处理24 h后的IC50作为作用浓度。在Transwell实验,加松花粉药液组细胞侵袭数低于未加松花粉药液组,差异有统计学意义(P<0.01)。在Western Blot实验中,ENO1蛋白和下游信号通路蛋白分子PI3K、AKT、C-MYC的表达量大部分呈现出加松花粉药液组低于未加松花粉药液组(P<0.05)。结论松花粉有抑制SMMC-7721肝癌细胞增殖和侵袭作用,并且其作用机制与抑制ENO1、PI3K、AKT和C-MYC的表达量有关。Objective To investigate the effect of pine pollen on the proliferation and invasion of hepatocellular carcinoma cell line SMMC-7721,and expression changes of the proteins of phosphatidylinositol 3-kinase/protein serine threonine kinase(PI3K/AKT)signaling pathway related to such effect.Methods Firstly,this study detected the concentrations of total polysaccharide,total flavonoids and zinc ions in the water extract of pine pollen.SMMC-7721 cells without pine pollen were taken as the control group.SMMC-7721 cells were treated with a series of concentrations of pine pollen for 12 h and 24 h,respectively.The CCK8 method was adopted to detect the proliferation and optimal semi inhibitory concentration(IC50)of cells.In addition,IC50 of SMMC-7721 cells was used as the concentration of the group treated with pine pollen.The cell invasion was detected with Transwell method.Western Blot was employed to detect the expression changes ofα-Enolase(ENO1)and the proteins in downstream PI3K/AKT signaling pathway.Results According to CCK8 detection,the IC50 of SMMC-7721 cells treated with pine pollen for 24 h was 16.78μg/μL,which was better than that treated with 23.87μg/μL for 12 h.Therefore,the IC50 of pine pollen treated for 24 h was used as the acting concentration in subsequent experiments.In Transwell experiment,the number of invaded cells in the groups with pine pollen liquid was lower than that in the group without pine pollen liquid,and the difference was statistically significant(P<0.01).In the Western Blot experiment,it turned out that most groups with pollen liquid had lower expressions of ENO1 and protein molecules(PI3K,AKT and C-MYC)in downstream signaling pathway than the group without pollen liquid(P<0.05).Conclusion Pine pollen can inhibit the proliferation and invasion of hepatoma cell line SMMC-7721,and its mechanism is related to the inhibition of ENO1,PI3K,AKT and C-MYC expressions.

关 键 词：α烯醇酶 松花粉 肝肿瘤 

分 类 号：R735.7[医药卫生—肿瘤]