Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025 Jun 18;30(12):2634.
doi: 10.3390/molecules30122634.

Alteration of m6A Methylation in Breast Cancer Cells by Kalanchoe pinnata Aqueous Extract

Affiliations

Alteration of m6A Methylation in Breast Cancer Cells by Kalanchoe pinnata Aqueous Extract

Carlos Rogelio Alvizo-Rodríguez et al. Molecules. .

Abstract

Kalanchoe pinnata is used in traditional medicine to treat cancer, as it contains flavonoids and phenols known to regulate key cellular processes associated with cancer. Breast cancer, the most common cancer among women globally, presents ongoing challenges in treatment. The discovery of m6A methylation and its regulation by methylosome proteins offers novel therapeutic avenues for cancer management. This study aimed to investigate the cytotoxic and epitranscriptomic effects of an aqueous extract from K. pinnata on MCF-7 (luminal A) and HCC1937 (triple-negative) breast cancer cells. Cell lines were treated with the aqueous K. pinnata extract, characterized by HPLC, for 72 h, followed by an assessment of cytotoxicity and migration. The expression of methylosome components METTL3 and FTO was measured using RT-PCR. m6A global methylation was assessed via colorimetry, and molecular docking studies were conducted. The results indicated that only HCC1937 cells exhibited altered migration capacity. This change was correlated in silico with the inhibition of METTL3 by luteolin and quercetin, constituents of the aqueous extract. METTL3, a methyltransferase, was overexpressed by scratch stimuli but was downregulated following K. pinnata treatment in both MCF-7 and HCC1937 cells. The FTO demethylase was overexpressed in both cell lines. In silico analysis suggested an interaction between FTO and compounds such as gallic acid and myricetin. Additionally, m6A global methylation decreased in MCF-7 cells but increased in HCC1937 cells, potentially affecting cell migration. Our findings indicate that K. pinnata influences both METTL3 and FTO, altering m6A methylation in a cell-type-dependent manner, with HCC1937 cells being particularly sensitive. Further research is required to elucidate the complete molecular mechanism of K. pinnata's aqueous extract in breast cancer treatment.

Keywords: FTO; HCC1937 cells; Kalanchoe pinnata; MCF-7 cells; METTL3; breast cancer; m6A; migration.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
Cytotoxic effects of aqueous extract of K. pinnata cells after 72 h of treatment. (A) MTT assay, (B) trypan blue assay. The statistical significance was determined through the calculation of the p-value where the mean ± SEM of percentage cell viability of the human breast cancer cells was compared using a t-test. The statistical significance is represented by asterisks (* p < 0.05; ** p < 0.01).
Figure 2
Figure 2
Effect of aqueous extract of K. pinnata on cell migration after 72 h of treatment. Microscopic images of wound healing assay in control and treated HCC1937 (A) and MCF-7 (B) cells. Photos were taken 0, 24, 48, and 72 h after scratch formation (scale bar equal to 100 µm). Percentage of wound closure in HCC1937 (C) and MCF-7 (D) cells. The statistical significance is represented by asterisks (* p < 0.05; ** p < 0.01) compared with the control without extract. The MCF-7 cells were treated with 40 µg/mL of aqueous extract, and the HCC1937 cells with 66 µg/mL.
Figure 3
Figure 3
Changes in METTL3 expression after treatment with aqueous extract of K. pinnata in (A) MCF-7 and (B) HCC1937 cells. A higher expression was observed in HCC1937 cells after the wound was performed but with no effect from the aqueous extract only. The non-treated group was the control (C), along with a wound healing group (W), an aqueous extract group (AE), and a wound healing and aqueous extract group (WAE) (* p < 0.05; ** p < 0.01, *** p < 0.001). The MCF-7 cells were treated with 40 µg/mL of aqueous extract, and the HCC1937 cells with 66 µg/mL.
Figure 4
Figure 4
Changes in FTO expression after treatment with aqueous extract of K. pinnata in (A) MCF-7 and (B) HCC1937 cells. The non-treated group was the control (C), along with a wound healing group (W), an aqueous extract group (AE), and a wound healing and aqueous extract group (WAE), (* p < 0.05; ** p < 0.01, *** p < 0.001). The MCF-7 cells were treated with 40 µg/mL of aqueous extract, and the HCC1937 cells with 66 µg/mL.
Figure 5
Figure 5
Effect of aqueous extract of K. pinnata on m6A global methylation. The m6A levels in (A) MCF-7 and (B) HCC1937 cells were detected using a m6A RNA kit. The data represent the average of three independent experiments (mean ± SEM). The non-treated group was the control (C), along with a wound healing group (W), an aqueous extract group (AE), and a wound healing and aqueous extract group (WAE), (* p < 0.05; ** p < 0.01, *** p < 0.001). The MCF-7 cells were treated with 40 µg/mL of aqueous extract, and the HCC1937 cells with 66 µg/mL.

Similar articles

Cited by

References

    1. Rajsekhar P., Bharani R., Ramachandran M., Angel K., Rajsekhar S. The “Wonder Plant” Kalanchoe pinnata (Linn.) Pers.: A Review. J. Appl. Pharm. Sci. 2016;6:151–158. doi: 10.7324/JAPS.2016.60326. - DOI
    1. Allorge-Boiteau L. Madagascar Centre de Speciation et D’origine du Genre Kalanchoe (Crassulaceae) Biogéographie Madagasca. 1996:137–145.
    1. Fernandes J.M., Cunha L.M., Azevedo E.P., Lourenço E.M.G., Fernandes-Pedrosa M.F., Zucolotto S.M. Kalanchoe laciniata and Bryophyllum pinnatum: An updated review about ethnopharmacology, phytochemistry, pharmacology and toxicology. Rev. Bras. Farmacogn. 2019;29:529–558. doi: 10.1016/j.bjp.2019.01.012. - DOI
    1. Elufioye T.O., Oyedeji A.O., Habtemariam S. A Review of the Traditional Uses, Phytochemistry and Pharmacology of Bryophyllum pinnatum (Lam.) (Crassulaceae) J. Biol. Act. Prod. Nat. 2022;12:190–222. doi: 10.1080/22311866.2021.1988706. - DOI
    1. Sung H., Ferlay J., Siegel R.L., Laversanne M., Soerjomataram I., Jemal A., Bray F. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J. Clin. 2021;71:209–249. doi: 10.3322/caac.21660. - DOI - PubMed

MeSH terms

Grants and funding

LinkOut - more resources