Toxicity and Anti-Proliferative Properties of Anisomeles indica Ethanol
Extract on Cervical Cancer HeLa Cells and Zebrafish Embryos
Nguyen T. Bich-Loan 1,2, Kieu Trung Kien 1, Nguyen Lai Thanh 1, Nguyen T. Kim-Thanh 1, Nguyen Quang Huy 1,Pham The-Hai, Marc Muller 3, Amandine Nachtergael 2, Pierre Duez 2 and Nguyen Dinh Thang 1,*
1 Faculty of Biology, VNU University of Science, Vietnam National University, Hanoi 100000, Vietnam;
nguyenthibichloan.iph@gmail.com (N.T.B.-L.); kieukien1@gmail.com (K.T.K.); nguyenlaithanh@hus.edu.vn (N.L.T.); kimthanh_bio@yahoo.com (N.T.K.-T.); huy_nq@hus.edu.vn (N.Q.H.); hai.phamthe@gmail.com (P.T.-H.)
2 Unit of Therapeutic Chemistry and Pharmacognosy, University of Mons (UMONS), 7000 Mons, Belgium; Amandine.NACHTERGAEL@umons.ac.be (A.N.); Pierre.DUEZ@umons.ac.be (P.D.)
3 Laboratory for Organogenesis and Regeneration, GIGA-R, Department Life Sciences, University of Liege, 4000 Liege, Belgium; m.muller@uliege.be
* Correspondence: ndthang@hus.edu.vn
Abstract: In this study, we showed that crude extract of Anisomeles indica (AI-EtE) expressed its toxicity to HeLa cells with an IC50 dose of 38.8 µg/mL and to zebrafish embryos with malformations, lethality and hatching inhibition at 72-hpf at doses higher than 75 µg/mL. More interestingly, flow cytometry revealed that AI-EtE significantly promoted the number of cells entering apoptotic. Accordingly, the transcript levels of BAX, CASPASE-8, and CASPASE-3 in the cells treated with AI-EtE at IC50 dose were 1.55-, 1.62-, and 2.45-fold higher than those in the control cells, respectively. Moreover, treatment with AI-EtE caused cell cycle arrest at the G1 phase in a p53-independent manner. Particularly, percentages of AI-EtE-treated cells in G1, S, G2/M were, respectively 85%, 6.7% and 6.4%; while percentages of control cells in G1, S, G2/M were 64%, 15% and 19%, respectively. Consistent with cell cycle arrest, the expressions of CDKN1A and CDNK2A in AI-EtE-treated cells were up-regulated 1.9- and 1.64-fold, respectively. Significantly, treatment with AI-EtE also decreased anchorage-independent growth of HeLa cells. In conclusion, we suggest that Anisomeles indica can be considered as a medicinal plant with a possible use against cervical cancer cells; however, the used dose should be carefully monitored, especially when applying to pregnant women.
Keywords: medicinal plant; Anisomeles indica; HeLa cell; zebrafish; apoptosis; cell cycle arrest; gene expression; colony formation
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