海南医科大学学报2025，Vol.31Issue(11)：824-832,839,10.DOI:10.13210/j.cnki.jhmu.20241021.003
基于PI3K/Akt、NF-κB信号通路探究黄精多糖对前列腺癌PC-3细胞裸鼠模型作用的研究
Study on the effect of polygonatum Sibiricum polysaccharides on nude mice model of prostate cancer PC-3 cells based on PI3K/Akt and NF-κB signaling pathways
摘要
Abstract
Objective:To investigate the impacts of Polygonatum sibiricum polysaccharides(PSP)on the growth of PC-3 cell transplanted tumors in nude mice and key proteins in the phosphatidylinositol-3-hydroxy kinase/protein kinase B(PI3K/Akt)and nuclear factor-κB(NF-κB)signaling pathways.Methods:PC-3 androgen-independent prostate cancer cell line was selected and in-oculated into the subcutaneous tissue of BALB/c nude mice to establish a model.The mice were divided into 5 groups,with 6 mice in each group,and the drug dosage was adjusted based on the conversion of body surface area between humans and nude mice.The control group was given physiological saline,cyclophosphamide(CTX)group was given 30 mg·kg-1·d-1,low-dose group was given PSP at 100 mg·kg-1·d-1,medium-dose group was given PSP at 200 mg·kg-1·d-1,high-dose group was given PSP at 400 mg·kg-1·d-1.Each nude mouse was administered 0.2 mL by gavage every other day for 30 days.Every three days,the body weight and tumor volume of the nude mice were recorded to analyze the effect of PSP on the growth and size of the transplanted tu-mors.Tumor volume was measured 4 hours after the last drug administration to calculate the tumor inhibition rate(TIR).Obser-vation of cell apoptosis in tumor tissue using TdT-mediated dUTP nick end labeling(TUNEL)and flow cytometry.The prolifera-tion activity of spleen cells was detected by Cell Counting Kit-8(CCK8).The activity of natural killer cells(NKc)was detected by lactate dehydrogenase(LDH)release assay.Using enzyme-linked immunosorbent assay(ELISA)to detect the levels of tumor necrosis factor-α,interleukin-2,and interleukin-6(TNF-α,IL-2,IL-6).Immunohistochemistry(IHC)was performed to analyze the expression of PI3K,Akt,NF-κB P65,and phosphorylated protein(p-PI3K,p-Akt,NF-κB p-P65)and caspase-3 in the transplanted tumor tissues of nude mice.Results:In the early stage of model establishment,nude mice were in good spirits,with free and frequent movement,stable food intake,and sensitive responses to external stimuli.However,with the growth of the tu-mor,all nude mice showed poor mental state,reduced activity and diet.Compared to the control group,TIR,apoptosis rate of PC-3 cells(ARoPc)in flow cytometry,and caspase-3 expression(IHC)increased in the CTX and PSP groups(P<0.05,P<0.001).ARoPc in TUNEL(CTX group),NK cell vitality(LDH),IL-6 levels(ELISA)in medium-and high-dose PSP groups,spleen cell proliferation(CCK8)in high-dose PSP,and TNF-α and IL-2 levels in PSP groups were also increased(P<0.05).Tumor volume and p-P65(IHC)in CTX and PSP groups,along with TNF-α and IL-2 levels(ELISA),p-PI3K,and p-Akt(IHC)in CTX,and p-PI3K and p-Akt(IHC)in medium-and high-dose PSP groups decreased(P<0.05).Compared to the CTX group,the PSP group showed increased tumor volume,NKc vitality(LDH),TNF-α,IL-2 levels(ELISA),and p-P65(IHC)(P<0.05).Spleen cell vitality(CCK8)and IL-6 levels(ELISA)in medium-and high-dose groups,as well as p-PI3K and p-Akt levels(IHC)in the low-dose PSP group,were also increased(P<0.05).Tumor IR and ARoPc(TUNEL,flow cytometry)in the PSP group,along with caspase-3 levels(IHC)in low-and medium-dose groups,were decreased(P<0.01,P<0.001).In medium-and high-dose PSP groups,tumor IR,ARoPc(TUNEL,flow cytometry),TNF-α,IL-2,IL-6 levels(ELISA),and caspase-3 levels(IHC)increased compared to the low-dose group(P<0.05),while tumor volume,p-PI3K,p-Akt(IHC)in these groups,and p-P65(high-dose)were decreased(P<0.05,P<0.001).The high-dose group showed increased tumor IR,ARoPc(TUNEL,flow cytometry),TNF-α,IL-2,IL-6 levels(ELISA),and caspase-3 levels(IHC)compared to the medium-dose group(P<0.05),with reductions in tumor volume and p-PI3K,p-Akt,and p-P65 levels(IHC)(P<0.05).Conclusion:PSP can inhibit the proliferation of PC-3 cells and induce their apoptosis.The mechanism may be related to inhibiting the PI3K/Akt and NF-κB signaling pathways by reducing the activation of PI3K,Akt,and P65 proteins,and promoting the expression of Caspase-3.It may be also related to regulating the activity of immune cells,and levels of TNF-α,IL-2,and IL-6.关键词
黄精多糖/PC-3细胞/裸鼠/PI3K/Akt/NF-κB P65Key words
Polygonatum sibiricum polysaccharides/PC-3 cells/Nude mice/PI3K/Akt/NF-κB P65分类
中医学引用本文复制引用
杨帅博,曾鑫,刘晨曦,唐玉红,周艳,董少文,赵欣阳,刘鑫,李晨,王新梅,赵国斌..基于PI3K/Akt、NF-κB信号通路探究黄精多糖对前列腺癌PC-3细胞裸鼠模型作用的研究[J].海南医科大学学报,2025,31(11):824-832,839,10.基金项目
河北省自然科学基金资助项目(H2021405012) This study was supported by Natural Science Foundation of Hebei Province(H2021405012) （H2021405012）
