参芍消瘤方含药血清对人肝癌细胞生物学行为的影响  

Effect of Shenshaoxiaoliu formula containing serum on biological behavior of human liver cancer cell

作　　者：丁晓明 张贤梅 柯亭 昝俊杰 孙勤国 DING Xiaoming;ZHANG Xianmei;KE Ting;ZAN Junjie;SUN Qinguo(Department of Traditional Chinese Medicine,Wuhan Third Hospital,Wuhan 430060,China)

机构地区：[1]武汉市第三医院中医科,430060

出　　处：《浙江医学》2023年第9期962-967,I0004,共7页Zhejiang Medical Journal

基　　金：第七批全国老中医药专家学术经验继承工作项目(国中医药人教函〔2022〕76号);武汉市卫生健康委员会基金资助项目(WZ21A05)。

摘　　要：目的探讨参芍消瘤方含药血清对人肝癌细胞Bel-7402增殖、凋亡、细胞周期等生物学行为的影响及其作用机制。方法使用SD大鼠制备参芍消瘤方含药血清。将Bel-7402分为对照组、含药血清组(25%含药血清)、顺铂组(5 mmol/L顺铂)、含药血清+顺铂组(25%含药血清+5 mmol/L顺铂)。采用5-乙炔基-2'脱氧尿嘧啶核苷(EdU)染色法检测细胞增殖情况;采用流式细胞术检测细胞周期和细胞凋亡率;采用划痕法检测细胞迁移能力;采用Transwell法检测细胞侵袭能力;采用Western blot法检测细胞磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)和半胱氨酸天冬氨酸蛋白酶3(Caspase-3)蛋白表达水平。结果与对照组比较,含药血清组和顺铂组细胞增殖数量减少,侵袭能力下降,磷酸化PI3K(p-PI3K)、磷酸化Akt(p-Akt)蛋白表达水平降低,划痕速度减慢,凋亡率、S期细胞占比升高,Caspase-3蛋白表达水平升高,差异均有统计学意义(均P<0.05)。与顺铂组比较,含药血清+顺铂组细胞增殖数量减少,侵袭能力下降,p-PI3K、p-Akt蛋白表达水平降低,划痕速度减慢,凋亡率、S期细胞占比升高,Caspase-3蛋白表达水平升高,差异均有统计学意义(均P<0.05)。结论参芍消瘤方含药血清可抑制Bel-7402细胞增殖、迁移、侵袭,诱导细胞周期阻滞,促进细胞凋亡,其作用机制可能与抑制PI3K/Akt信号通路有关。Objective To study the effects and mechanism of drug-containing serum of Chinese medicine Shenshaoxiaoliu formula on the proliferation,apoptosis,cell cycle and other biological behaviors of human liver cancer cells.Methods The Shenshaoxiaoliu formula medicated serum was prepared in SD rats.Human hepatoma Bel-7402 cells divided into control group,medicated serum group(treated with 25%medicated serum)and CDDP group(treated with 5 mmol/L cisplatin),medicated serum+CDDP group(treated with 25%medicated serum+5 mmol/L cisplatin).Cell proliferation was detected by 5-ethynyl-2'-deoxyuridine(EdU)staining;cell cycle and apoptosis rate were detected by flow cytometry;cell migration was detected by scratch method,cell invasion was detected by Transwell method.The protein expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(Akt)and cysteine aspartate protease 3(Caspase-3)in Bel-7402 cells were detected by Western blot.Results Compared with the control group,the cell proliferation,invasion and expression levels of p-PI3K and p-Akt in the medicated serum group and the CDDP group were significantly decreased,while the scratch rate slowed down,and the apoptosis rate,proportion of S phase cells and the Caspase-3 protein expression level were significantly increased(all P<0.05).Compared with the CDDP group,the cell proliferation,invasion and expression levels of p-PI3K and p-Akt were significantly decreased in medicated serum+CDDP group,while the rate of scratch slowed down,and the apoptosis rate,the proportion of S phase cells,and the Caspase-3 protein expression level were significantly increased(all P<0.05).Conclusion The medicated serum of Chinese medicine Shenshaoxiaoliu formula can inhibit the proliferation,migration and invasion of Bel-7402 cells,induce cell cycle arrest,and promote cell apoptosis,which may be related to the inhibition of PI3K/Akt signaling pathway.

关 键 词：参芍消瘤方 肝癌 增殖 凋亡 PI3K/AKT信号通路 

分 类 号：R285[医药卫生—中药学]