Abstract
Objective To explore the mechanism of action of Fuzheng Yiliu formula (FZYLF) in regulation of the invasion and metastasis of MDA-MB-231/Adr human breast cancer cells through WAVE3. Methods The MDA-MB-231/Adr cells with high invasive ability were screened by Transwell, and the plasmid with high WAVE3 expression was made for transfection. Plasmid transfection efficiency and protein expression level were verified by polymerase chain reaction (PCR) and western blotting (WB). The effect of FZYLF on cell proliferation and invasion was investigated before and after WAVE3 silencing by flow cytometry. A nude mouse model of tumor metastasis was established to study the antitumor activity of FZYLF. Results The expression levels of mRNA and proteins of intracellular WAVE3 increased significantly after plasmid transfection, mRNA from 1.37± 0.41 to 9.88 ± 1.31 and protein from 1 ± 0.08 to 5.09 ± 0.03 (P < 0.01). Intervention with FZYLF could significantly affect the activity of MDA-MB-231/Adr cells and inhibit invasion and metastasis, IC50 from 71.04 to 46.41 mg/mL and from 162 ± 14.82 to 81.4 ± 12.05 (P < 0.05 or P < 0.01), and significantly reduce the expression levels of WAVE3 (from 1 ± 0.02 to 0.63 ± 0.04), MMP-9 (from 1 ± 0.05 to 0.63 ± 0.03), NF-κB (p65) (from 1 ± 0.02 to 0.62 ± 0.02), and p-IκBα (from 1 ± 0.03 to 0.68 ± 0.02) (P < 0.05 or P < 0.01). The T/C (%) of FZYLF (13 g crude drug/kg) was 62.06% for MDA-MB-231/Adr tumor xenografted in nude mice, with a tumor inhibition rate of 39.64%. Conclusion FZYLF can inhibit the invasion and proliferation of the MDA-MB-231/Adr human breast cancer cells, and the mechanism of action may be related to the regulation of WAVE3 expression.
Highlights
Results (Figure 2) showed that the expression level of WAVE3 was higher in the drug-resistant MDA-MB-231/Adr and MDA-MB-231/ Taxol cells than in the nondrug-resistant MDA-MB231 cells, and it was higher in the drug-resistant MCF-7/Adr and MCF-7/Taxol cells than in the nondrug-resistant MCF7 cells. e expression level of WAVE3 was the highest in the MDA-MB-231/Adr cells
Results (Figure 3) showed that the mRNA and protein expression levels of WAVE3 increased significantly in the MDA-MB-231 cells transfected with pcDNA3.1-WAVE3 plasmid (P < 0.01), and Figure 4 shows that the levels decreased significantly in the MDA-MB-231/Adr cells with WAVE3 knocked out (P < 0.05 or P < 0.01)
Epithelial cells of the breast that result in different groups of tumor cells. rough analysis of the molecular markers by immunohistochemistry, breast cancer is classified into four types based on different expressions of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor-2 (HER-2), and Ki67 index [19,20,21]
Summary
Breast cancer is one of the most common malignant tumors in women all over the world and has gradually evolved into the first female malignant tumor in recent years [1]. is disease tends to recur and metastasize and its biological behavior is dangerous, especially in triple negative breast cancer (TNBC), which has the characteristics of poor histological grade, advanced clinical stage, poor prognosis and high mortality, and a tendency to occur at a younger age. e 3- and 5-year disease-free survival (DFS) and overall survival (OS) are lower in TNBC than in non-TNBC [2]. e prognosis of TNBC gets worse with the increase in N staging, and the probability of recurrence and metastasis increases significantly at stage N2 or above, but there is no significant correlation with T staging and histological grade [3]. WAVE3 plays an important role in migration and invasion of breast cancer cells. The expression of WAVE3 is positively correlated with the grade, drug resistance, invasion, and progression of tumor. Silencing the expression of WAVE3 in invasive breast cancer cells by siRNA can effectively reduce the number of lamellar pseudopodia on the edge of tumor cell membrane and weaken the ability of tumor cells to invade and migrate. After the breast cancer cells with further knockout of WAVE3 are inoculated into nude mice, it is found out that compared to the negative control group, the growth of tumor in situ is significantly slower and the distant metastasis rate of tumor decreases significantly [8]. Ese results suggest that WAVE3 plays an important role in the invasion and metastasis of breast cancer, especially in chemotherapyresistant breast cancer It has been confirmed that the expression of WAVE3 is higher in tissues of invasive breast cancer than in tissues of breast cancer in situ [10]. ese results suggest that WAVE3 plays an important role in the invasion and metastasis of breast cancer, especially in chemotherapyresistant breast cancer
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
