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Mycosubtilin Induces G1 Phase Block and Autophagy in Cervical Cancer HeLa Cells

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Abstract

Cyclic lipopeptides secreted by the probiotic bacterium Bacillus subtilis have attracted much attention due to their antitumor activities and low toxicity. However, the role of Mycosubtilin (Myco) in the prevention and treatment of cervical cancer remains unclear. In the present study, we conducted a systematic evaluation of Myco’s anti-cervical cancer effects to identify its molecular mechanism of action using proteomics technology. The results reveal that Myco inhibited the growth of HeLa and SiHa cervical cancer cell lines in a dose-dependent (3–15 µg/mL) and time-dependent (12–48 h) manner and significantly reduced colony formation and migration in HeLa cells, highlighting its potential to suppress tumor spread. Moreover, autophagosome and autolysosome numbers were significantly increased after Myco treatment, and the expression of autophagy-related proteins was significantly modulated, suggesting that autophagy plays a role in its anti-cancer mechanism. Myco treatment also induced G1 phase cell cycle arrest in HeLa cells, as confirmed by proteomics analysis. Myco was shown to induce cell cycle arrest in HeLa cells by regulating the P53 pathway and autophagy-dependent cell death via the PI3K/AKT/mTOR signaling pathway, demonstrating its multidimensional effect on cervical cancer cell growths. Myco treatment significantly inhibited tumor growth in vivo in a nude mouse cervical cancer xenograft model, providing direct evidence of its potential as a therapeutic candidate for cervical cancer. Given its unique anti-cancer mechanism and significant therapeutic efficacy, Myco should be considered a promising therapeutic agent for cervical cancer.

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Data Availability

No datasets were generated or analysed during the current study.

Abbreviations

Myco:

Mycosubtilin

DMSO:

Dimethyl sulfoxide

HPLC:

High-performance liquid chromatography

DMEM:

Dulbecco’s modified eagle medium

RPMI:

Roswell park memorial institute

CCK- 8:

Cell counting kit-8

G1:

First gap

S:

Synthesis

G2:

Second gap

M:

Metaphase

BS-Z15:

Bacillus subtilis-Z15

H&E:

Hematoxylin and eosin

OD:

Optical density

PBS:

Phosphate-buffered saline

PI:

Propidium iodide

RNase A:

Ribonuclease A

MDC:

Monodansulfonyl cadaverine

3-MA:

3-Methyladenine

RIAP:

Radio immunoprecipitation assay

BCA:

Bicinchoninic acid

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

PVDF:

Polyvinylidene fluoride

TBST:

Tris-buffered saline and Tween 20

ECL:

Enhanced chemiluminescence

IC50 :

Half maximal inhibitory concentration

DDP:

Diamminedichloroplatinum

KEGG:

Kyoto Encyclopedia of Genes and Genomes

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Funding

Natural Science Foundation of China,No.32160074,Open project of Key Laboratory in Xinjiang,No.2020D4010

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Authors and Affiliations

  1. Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology, College of Life Science, Xinjiang Normal University, Urumqi, China

    Haoran Li, Dongyuan Zhou, Weiquan Wang, Reyihanguli Aimaier,  JunYang, Haoran Zhao, Jinyu Li, Qian Zhou & Huixin Zhao

  2. Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, China

    Heping Zhao

  3. Shanghai Key Laboratory of Regulatory Biology and School of Life Sciences, East China Normal University, Shanghai, 200241, China

    Xiufeng Pang

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  1. Haoran Li
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Contributions

Conceptualization, Huixin Zhao, Qian Zhou and Xiufeng Pang; Data curation, Heping Zhao, Reyihanguli Aimaier and JunYang; Formal analysis, Dongyuan Zhou, Qian Zhou and Huixin Zhao; Funding acquisition,Huixin Zhao; Investigation, Haoran Li and Weiquan Wan; Methodology,Jinyu Li, Haoran Li; Project administration, Huixin Zhao; Resources, Huixin Zhao; Supervision, Xiufeng Pang; Writing-original draft, Haoran Li and Dongyuan Zhou; Writing -review & editing, Huixin Zhao.Huixin Zhao is the first corresponding author, and Qian Zhou and Professor Xiufeng Pang are the co corresponding authors. Haoran Li, Dongyuan Zhou and Weiquan Wang these authors contributed equally to this work and share first authorship. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Xiufeng Pang, Qian Zhou or Huixin Zhao.

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Li, H., Zhou, D., Wang, W. et al. Mycosubtilin Induces G1 Phase Block and Autophagy in Cervical Cancer HeLa Cells. Probiotics & Antimicro. Prot. (2025). https://doi.org/10.1007/s12602-025-10534-1

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