抗癌扶正方通过调控自噬及JAK2/STAT3信号通路抑制肝细胞癌进展实验研究
Anti-cancer Fuzheng Formula Inhibits Hepatocellular Carcinoma Progression by Regulating Autophagy and JAK2/STAT3 Signaling Pathway
许婉丹 1程小波 2王春雷3
作者信息
- 1. 浙江中医药大学 浙江 杭州 310053;临海市中医院 浙江 临海 317000
- 2. 临海市中医院 浙江 临海 317000
- 3. 浙江省肿瘤医院 浙江 杭州 310022
- 折叠
摘要
目的:探究抗癌扶正方调控细胞自噬对肝细胞癌(hepatocellular carcinoma,HCC)进展的影响.方法:人肝癌细胞SMMC-7721给予抗癌扶正方(0、1、2mg/mL)、顺铂(4mg/L)分别处理,分为对照组、抗癌扶正方低、高剂量组、顺铂组.同时,将SMMC-7721细胞分为抗癌扶正方组(2mg/mL)、抗癌扶正方(2mg/mL)+3-甲基腺嘌呤(3-Methyladenine,3-MA,2mmol/L)组、对照组(等量生理盐水).按分组给药后于细胞培养箱中培养48h.MTT、流式MDC检测SMMC-7721细胞抑制率和自噬率情况.Western blot检测SMMC-7721细胞中自噬及JAK2/STAT3信号通路相关蛋白表达水平.结果:与对照组相比,抗癌扶正方低、高剂量组和顺铂组对SMMC-7721细胞的抑制率和自噬率显著升高,Beclin-1、LC3蛋白表达显著增加,P62蛋白表达显著降低(P＜0.05);且抗癌扶正方高剂量组SMMC-7721细胞的P-JAK-2/JAK-2、P-STAT3/STAT3蛋白表达显著降低(P＜0.01).而与抗癌扶正方相比,抗癌扶正方+3-MA组细胞抑制率和自噬率则显著下降(P＜0.01).结论:抗癌扶正方可上调Beclin-1、LC3蛋白,下调P62蛋白以增强细胞自噬,并通过抑制JAK2/STAT3信号通路,共同干预HCC进展.
Abstract
Objective:This study aimed to investigate the effect of anti-cancer FuZheng formula to regulate cell autophagy on the progression of hepatocellular carcinoma(HCC).Methods:Human hepatoma cells SMMC-7721 were treated with anti-cancer FuZheng formula(0,1,2 mg/mL)and cisplatin(4 mg/L),respectively,and divided into control group,anti-cancer Fuzheng formula low and high dose group,and cisplatin group.Meanwhile,SMMC-7721 cells were divided into the anti-cancer Fuzheng formula group(2 mg/mL),anti-cancer Fuzheng formula(2 mg/mL)+ 3-methylad-enine(3-Methyladenine,3-MA,2 mmol/L)group,and the control group(equal amount of saline).And incubated in the cell culture incubator for 48 h after administration of drugs according to the group.MTT,flow MDC assay were performed to detect inhibition rate and autophagy rate of SMMC-7721 cells.Western blot was performed to detect the expression levels of autophagy-related molecu-lar proteins and JAK2/STAT3 signaling pathway-related proteins in SMMC-7721 cells.Results:Com-pared with the control group,the inhibition rate and autophagy rate of SMMC-7721 cells in the low and high dose groups of anti-cancer Fu ZhengFang formula and cisplatin group were signifi-cantly higher,and the expression of Beclin-1 and LC3 proteins were significantly increased and P62 protein expression was significantly decreased(P＜0.05);SMMC-7721 cells in the high dose group of anti-cancer FuZheng formula had significantly higher P-JAK-2/JAK-2 and P-STAT3/STAT3 protein expression was significantly decreased in SMMC-7721 cells at high dose(P＜0.01).While the inhibition rate and autophagy rate of SMMC-7721 cells were significantly lower in the Anti-Cancer Fu Zheng formula + 3-MA group compared with the anti-Cancer Fu Zheng formula(P＜0.01).Conclusion:Anti-cancer Fuzheng Formula up-regulates Beclin-1 and LC3 proteins,down-regulates P62 protein to enhance cellular autophagy,and inhibits JAK2/STAT3 signaling path-way,together interfering with HCC progression.
关键词
抗癌扶正方/肝细胞癌/自噬/JAK2/STAT3/细胞实验Key words
anti-cancer Fuzheng formula/hepatocellular carcinoma/autophagy/JAK2/STAT3引用本文复制引用
基金项目
浙江省自然科学基金/省药学会联合基金(LYY20H280001)
出版年
2024
国家科技期刊平台
NSTL
万方数据