Yiqi Yangjing recipe stimulates apoptosis while suppressing the energy metabolism via under-expression of PFKFB3 in A549 cells

Yu-Han Wang; Qing-Ting Wang; Xiao-Ting Wu; Yun Dong; Qing Cui; Ya-Ning Zhou; Xi-Wen Yang; Wen-Feng Lu; Wan-Yu Li; Hui Wang; Xiao-Dong Zhao; Ming Zhang

doi:10.21037/jtd-23-490

Yiqi Yangjing recipe stimulates apoptosis while suppressing the energy metabolism via under-expression of PFKFB3 in A549 cells

J Thorac Dis. 2023 Sep 28;15(9):4885-4895. doi: 10.21037/jtd-23-490. Epub 2023 Sep 21.

Authors

Yu-Han Wang^#¹, Qing-Ting Wang^#², Xiao-Ting Wu², Yun Dong¹, Qing Cui¹, Ya-Ning Zhou¹, Xi-Wen Yang³, Wen-Feng Lu⁴, Wan-Yu Li², Hui Wang², Xiao-Dong Zhao^#⁵, Ming Zhang^#¹

Affiliations

¹ Department of Integrated Traditional Chinese and Western Medicine, Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
² Shanghai Chest Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
³ Shanghai Literature Institute of Traditional Chinese Medicine, Shanghai, China.
⁴ Department of Integrated Traditional Chinese and Western Medicine, Zhongshan Hospital, Fudan University, Shanghai, China.
⁵ Shanghai Center for Systems Biomedicines, Shanghai Jiao Tong University, Shanghai, China.

^# Contributed equally.

Abstract

Background: Lung cancer is a malignant tumor associated with high morbidity and mortality. Yiqi Yangjing recipe (YYR) is a formula of traditional Chinese medicine (TCM) that is commonly used for the treatment of lung cancer with good clinical efficacy. The specific anti-cancer mechanism of YYR is still unknown. We need to embark on a more in-depth pharmacological study of YYR to determine the complex compound ingredients, which could be promoted in clinical practice to achieve efficacy in prolonging recurrent metastasis of lung cancer.

Methods: The cytotoxic effects of YYR on A549 cells were evaluated by Cell Counting Kit-8 (CCK-8) assay. The PFKFB3-under-expressed and overexpressed A549 cell lines were constructed via PFK15 treatment and transfection, respectively. The effects of YYR on PFKFB3 messenger RNA (mRNA) and protein expression were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot. The pro-apoptotic and anti-glycolytic abilities of YYR were measured using flow cytometry assay and hippocampal XF96 extracellular flux analyzer. An in vivo tumorigenicity assay was performed on nude mice to confirm the anti-cancer effects of YYR.

Results: YYR has a noticeable cytotoxic activity on A549 cells, with the treatment with both YYR and PFK15 significantly inducing apoptosis. YYR and PFK15 treatment reduced the extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) in A549 cells. Similar to PFK15, YYR can down-regulate PFKFB3 expression, and PFKFB3 overexpression suppressed the apoptosis, which was reversed by YYR. Animal experiments confirmed that YYR was able to inhibit tumor growth, induce tumor cell apoptosis, and down-regulate PFKFB3 in tumor tissues.

Conclusions: This study demonstrated that YYR promoted lung cancer cell apoptosis and inhibited energy metabolism by targeting PFKFB3. Furthermore, we believe that YYR may be a suitable supplement or alternative drug for lung cancer treatment.

Keywords: Apoptosis; PFKFB3; Yiqi Yangjing recipe (YYR); glycolysis; lung cancer.