Chen Wen-Li, Bai Huan-Huan, Liu Li-Wei, Chen Hong-Yu, Shi Qi, Chang Li-Sheng, Gou Xiao-Jun, Qian Jun
Baoshan District Hospital of Integrated Traditional Chinese and Western Medicine of Shanghai, Shanghai 201999, China.
School of Pharmacy, Shaanxi University of Traditional Chinese Medicine, Xianyang, Shaanxi 712046, China.
Evid Based Complement Alternat Med. 2021 Mar 27;2021:8898668. doi: 10.1155/2021/8898668. eCollection 2021.
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.
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.
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 ( < 0.01). Intervention with FZYLF could significantly affect the activity of MDA-MB-231/Adr cells and inhibit invasion and metastasis, IC from 71.04 to 46.41 mg/mL and from 162 ± 14.82 to 81.4 ± 12.05 ( < 0.05 or < 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-B (p65) (from 1 ± 0.02 to 0.62 ± 0.02), and p-IB (from 1 ± 0.03 to 0.68 ± 0.02) ( < 0.05 or < 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%.
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.
探讨扶正抑瘤方（FZYLF）通过WAVE3调控MDA-MB-231/Adr人乳腺癌细胞侵袭和转移的作用机制。
通过Transwell筛选出具有高侵袭能力的MDA-MB-231/Adr细胞，并构建高表达WAVE3的质粒进行转染。采用聚合酶链反应（PCR）和蛋白质免疫印迹法（WB）验证质粒转染效率和蛋白表达水平。通过流式细胞术研究WAVE3沉默前后FZYLF对细胞增殖和侵袭的影响。建立肿瘤转移裸鼠模型，研究FZYLF的抗肿瘤活性。
质粒转染后细胞内WAVE3的mRNA和蛋白表达水平显著升高，mRNA从1.37±0.41升高至9.88±1.31，蛋白从1±0.08升高至5.09±0.03（P<0.01）。FZYLF干预可显著影响MDA-MB-231/Adr细胞活性，抑制侵袭和转移，IC从71.04降至46.41mg/mL，侵袭细胞数从162±14.82降至81.4±12.05（P<0.05或P<0.01），并显著降低WAVE3（从1±0.02降至0.63±0.04）、基质金属蛋白酶-9（MMP-9，从1±0.05降至0.63±0.03）、核因子-κB（NF-κB，p65，从1±0.02降至0.62±0.02）和磷酸化IκB（p-IκB，从1±0.03降至0.68±0.02）的表达水平（P<0.05或P<0.01）。FZYLF（13g生药/kg）对裸鼠移植的MDA-MB-231/Adr肿瘤的T/C（%）为62.06%，肿瘤抑制率为39.64%。
FZYLF可抑制MDA-MB-231/Adr人乳腺癌细胞的侵袭和增殖，其作用机制可能与调控WAVE3表达有关。
