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Casticin Attenuates Stemness in Cervical Cancer Stem-Like Cells by Regulating Activity and Expression of DNMT1

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Abstract

Objective

To explore whether casticin (CAS) suppresses stemness in cancer stem-like cells (CSLCs) obtained from human cervical cancer (CCSLCs) and the underlying mechanism.

Methods

Spheres from HeLa and CaSki cells were used as CCSLCs. DNA methyltransferase 1 (DNMT1) activity and mRNA levels, self-renewal capability (Nanog and Sox2), and cancer stem cell markers (CD133 and CD44), were detected by a colorimetric DNMT activity/inhibition assay kit, quantitative real-time reverse transcription-polymerase chain reaction, sphere and colony formation assays, and immunoblot, respectively. Knockdown and overexpression of DNMT1 by transfection with shRNA and cDNA, respectively, were performed to explore the mechanism for action of CAS (0, 10, 30, and 100 nmol/L).

Results

DNMT1 activity was increased in CCSLCs compared with HeLa and CaSki cells (P<0.05). In addition, HeLa-derived CCSLCs transfected with DNMT1 shRNA showed reduced sphere and colony formation abilities, and lower CD133, CD44, Nanog and Sox2 protein expressions (P<0.05). Conversely, overexpression of DNMT1 in HeLa cells exhibited the oppositive effects. Furthermore, CAS significantly reduced DNMT1 activity and transcription levels as well as stemness in HeLa-derived CCSLCs (P<0.05). Interestingly, DNMT1 knockdown enhanced the inhibitory effect of CAS on stemness. As expected, DNMT1 overexpression reversed the inhibitory effect of CAS on stemness in HeLa cells.

Conclusion

CAS effectively inhibits stemness in CCSLCs through suppression of DNMT1 activation, suggesting that CAS acts as a promising preventive and therapeutic candidate in cervical cancer.

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Data Availability Statement

The data used to support the findings of this study are available from the corresponding author upon request.

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Authors and Affiliations

  1. Medical College, Hunan University of Medicine, Huaihua, Hunan Province, 418000, China

    Xue-li Wang & Kai-yu Deng

  2. Clinical Department of Guangdong Metabolic Disease Research Centre of Integrated Chinese and Western Medicine, the First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou, 510062, China

    Xiao-zheng Cao & Shao-qiang Lin

  3. Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China

    Xiao-zheng Cao & Yong Xu

  4. The Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Medical College, Hunan Normal University, Changsha, 410013, China

    Dao-yuan Wang, Ye-bei Qiu, Jian-guo Cao &  Kai-qun Ren

  5. The Key Laboratory of Study and Discover of Small Targeted Molecules of Hunan Province, Medical College, Hunan Normal University, Changsha, 410013, China

    Dao-yuan Wang, Ye-bei Qiu & Jian-guo Cao

Authors
  1. Xue-li Wang
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Contributions

Wang XL performed and analyzed the experiments, wrote the paper. Cao XZ, Wang DY, Qiu YB, Deng KY, Cao JG, Lin SQ and Xu Y carried out the data collection and data analysis. Ren KQ conceived and coordinated the study, participated in data analysis, and revised the paper. All authors contributed to data analysis, drafting or revising the article, gave final approval of the version to be submitted, and agree to be accountable for all aspects of the work.

Corresponding author

Correspondence to Kai-qun Ren.

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The authors declare that there is no conflict of interest.

Additional information

Supported by the National Natural Science Foundation of China (No. 82074075), Scientific Research Fund of Hunan Provincial Education Department (No. 20C1340), the Key Laboratory of Study and Discovery of Small Targeted Molecules of Hunan Province, Hunan Normal University (No. 2017TP1020), and the Huxiang High-Level Talent Innovation Team (No. 2018RS3072) 1. Medical College, Hunan University of Medicine, Huaihua, Hunan Province (418000), China; 2. Clinical Department of Guangdong Metabolic Disease Research Centre of Integrated Chinese and Western Medicine, the First Affiliated Hospital of Guangdong Pharmaceutical University, Guangzhou (510062), China; 3. Institute of Chemical Biology, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou (510530), China; 4. The Key Laboratory of Model Animals and Stem Cell Biology in Hunan Province, Medical College, Hunan Normal University, Changsha (410013), China; 5. The Key Laboratory of Study and Discover of Small Targeted Molecules of Hunan Province, Medical College, Hunan Normal University, Changsha (410013), China

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Wang, Xl., Cao, Xz., Wang, Dy. et al. Casticin Attenuates Stemness in Cervical Cancer Stem-Like Cells by Regulating Activity and Expression of DNMT1. Chin. J. Integr. Med. 29, 224–232 (2023). https://doi.org/10.1007/s11655-022-3469-z

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