摘要
Abstract
Background and purpose:Triple-negative breast cancer(TNBC)is characterized by strong invasiveness,high recurrence rate,and poor prognosis.Due to the lack of effective targeted therapies,there is an urgent need to find new,efficient therapeutic drugs in clinical practice.Psoralea corylifolia L.is a traditional Chinese medicine,and its active component corylifol A(CA)has been proven to have various biological activities such as anti-inflammatory and antioxidant effects.This study aimed to further investigate the role of CA in TNBC growth and its underlying mechanisms.Methods:The cell counting kit-8(CCK-8)was used to detect the effects of different concentrations of CA on the viability of various breast cancer cells.Evaluation of the impact of CA on the long-term proliferation capacity of TNBC cells through cloning experiments.Flow cytometry was used to quantitatively analyze CA-induced apoptosis.A 4T1 cell mouse subcutaneous transplant tumor model was established to evaluate the antitumor efficacy and safety of CA in vivo,and tumor tissue pathological damage was observed using hematoxylin-eosin staining(HE).Lysosomal function and autophagosome formation were observed using fluorescent probes and immunofluorescence staining,combined with the autophagy inhibitor chloroquine(CQ).Cloning experiments and Western blot analysis clarified the functional role of CA-induced autophagy in tumor cell survival.Finally,Western blot was used to detect the expression levels of autophagy markers,protein tyrosine phosphatase non-receptor type 1(PTPN1),and phosphorylation levels of the mammalian target of rapamycin(mTOR)signaling pathway,and the SwissTargetPrediction platform was used to predict potential CA targets.Molecular docking simulations were used to analyze the potential binding patterns between CA and PTPN1.The animal experiments in this study were approved by the Animal Experiment Project Ethics Review of the Animal Science Department,Experimental Center,Fudan University Shanghai Cancer Center(Approval No.FUSCC-IACUC-2026226).Results:CA inhibits the growth of TNBC cells,with the strongest inhibitory effects on SUM149 and MDA-MB-231,with an IC50 of approximately 10 μmol/L;meanwhile,flow cytometry analysis showed that CA treatment induces concentration-dependent apoptosis(P<0.05).In vivo experiments showed that CA(20 mg/kg)can slow the growth rate of mouse syngeneic tumor(P<0.01),leading to extensive necrosis in tumor tissues without affecting mouse body weight.Mechanistic studies found that CA treatment significantly increased the LC3-Ⅱ/Ⅰ ratio in TNBC cells(P<0.01),reduced the expression level of p62 protein(P<0.01),and was accompanied by increased LC3 fluorescence(P<0.01),suggesting that CA may promote autophagic flux.Lysosome staining experiments indicated that CA can cause a decrease in lysosomal fluorescence(P<0.01),suggesting that the acidic state or function of lysosomes may be altered.Functional reversal experiments showed that co-treatment with CQ further blocks autophagic response,enhancing the inhibitory effect on TNBC.Molecular docking results suggest that CA may form a relatively stable binding conformation with the PTPN1 protein,and Western blot showed that CA could downregulate PTPN1 protein expression(P<0.01)and reduce the phosphorylation level of the mTOR signaling pathway(P<0.05).Conclusion:CA can inhibit the growth of TNBC cells and promote cell apoptosis.Its anti-tumor effect may be related to the regulation of the PTPN1/mTOR signaling pathway and the induction of an autophagic response with protective tendencies.关键词
次苷酸查尔酮/三阴性乳腺癌/自噬/PTPN1/细胞凋亡Key words
Corylifol A/Triple-negative breast cancer/Autophagy/PTPN1/Apoptosis分类
医药卫生
