| In 2021,the World Health Organization reported that breast cancer(BC)had became the most common cancer worldwide.Chemotherapy as the main therapeutic strategy can lead to drug resistance,recurrence and metastasis.Metastasis is the main factor limiting the survival BC patient,so it is urgent to explore new drugs for the treatment of breast cancer metastasis(BCM).Cetyltrimethylammonium Bromide(CTAB)has played a therapeutic role in apoptosis of hepatoma cells,but its role in BCM remains unknown.This work aims to explore the inhibitory effect of CTAB on BCM and explore its mechanism.OD values of cells treated with different concentrations of CTAB for 24 h and 48 h were detected by MTT to explore the impact of CTAB on the viability of BC cells(MCF-7,MDA-MB-231,ZR-75-1 and 4T1)and screen the maximum non-toxic concentration of CTAB.Next,using the Wound-healing and Transwell migration experiments,the impact of CTAB on BC cells migration was studied in vitro.The impact of CTAB on lung metastasis of breast cancer in vivo: BALB /c mice with orthotopic lung metastasis of breast cancer were then established.Normal saline and CTAB were respectlively injected into the caudal vein every 3 days,meanwile weighing the body weight of the mice.30 days after inoculation with4T1 cells,and H&E staining was performed on lung sections.Further more,investigating the mechanism of CTAB.The influence of CTAB on breast cancer stem cells(BCSCs)was investigated by tumor microsphere formation experiment,immunofluorescence,flow cytometry,q RT-PCR and WB what is detected the content of CD44 and CD133.Secondly,we would using q RT-PCR and WB methods detect the expression of E-cadherin,N-cadherin,Vimentin and EMT-TFs(Snail and Twist)in control group(CON)and CTAB group(CTAB)to see what the effects of CTAB on BC cells and mouse EMT.Then,WB was used to detect the activation of AMPK by CTAB,Compound C(Com C),an inhibitor of AMPK,was used to perform Wound-healing and Transwell migration experiments and detect the expression of EMT-related proteins to further explore whether CTAB regulates AMPK and thus affects cell migration and EMT.Finally,ALT,AST and BUN were detected to verify whether CTAB has certain system safety.In this study,it is found that CTAB inhibits the migration of BC cells and reduces the number of lung metastatic nodules and the size of the lesions in mice.Following,CTAB inhibits CSCs dryness and reduced the proportion of CSCs,and down-regulates the expression of CSCs markers CD44 and CD133.Further studies showes that CTAB increases the expression of E-cadherin in BC cells and mice,down-regulates the expression of N-cadherin,Vimentin,Snailer and Twist.In addition,CTAB activates AMPK to inhibit EMT progression and cell migration.Finally,there are no significant differences in body weight,ALT,AST and BUN between CON group and CTAB group.Based on above-mentioned results,it was found that CTAB is able to inhibit the migration of BC cells and lung metastasis in mice.Mechanism study revealed that CTAB could inhibit the progression of EMT in vitro via activation AMPK,and affect migration through regulation of BSCSs.In addition,inhibition of EMT reduces lung metastasis of BC in vivo.CTAB is a promising agent in the clinical treatment of BC metastasis. |
