Objective:"Multi-targeting"drugs can prove fruitful to combat drug-resistance of multifactorial disease-cervical cancer.This study envisioned to reveal if Thuja homeopathic mother tincture(MT)and its bioac-tive component could combat human papillomavirus(HPV)-16-infected SiHa cervical cancer cells since it is globally acclaimed for HPV-mediated warts.Methods:Thuja MT was studied for its antiproliferative and antimigratory properties in SiHa cells fol-lowed by microscopic determination of reactive oxygen species(ROS)generation by 2′,7′-dichlorodihydro fluorescein diacetate(DCFDA)staining and loss in mitochondrial membrane potential(MtMP)by rho-damine 123(Rh123)staining.Apoptosis and autophagy inductions were studied by acridine orange/ethidium bromide(AO/EB)staining and immunoblot analyses of marker proteins.The bioactive compo-nent of Thuja MT detected by gas chromatography-mass spectrometry was studied for antiproliferative and antimigratory properties along with in silico prediction of its cellular targets by molecular docking and oral drug forming competency.Results:Thuja MT showed significant antiproliferative and antimigratory potential in SiHa cells at a 50％inhibitory concentration(IC50)of 17.3 μL/mL.An increase in DCFDA fluorescence and loss in Rh123 flu-orescence prove that Thuja MT acted through the burst of ROS and loss in MtMP respectively.AO/EB-stained cells under the microscope and immunoblot analyses supported Thuja-induced cellular demise via dual pathways-apoptosis and autophagy.Immunoblots showed cleavage of caspase-3 and poly(ade-nosine diphosphate-ribose)polymerase-1(PARP-1)along with upregulation of Beclin-1,microtubule-associated protein 1 light chain 3B(LC3B)-Ⅱ,and p62 proteins.Hence,the apoptotic cascade followed a caspase-3-dependent pathway supported by PARP-1 cleavage,while autophagic death was Beclin-1-dependent and mediated by accumulation of LC3BⅡ and p62 proteins.Thujone,detected as the bioactive principle of Thuja MT,showed greater anti-proliferative and anti-migratory potential at an IC50 of 77 μg/mL,along with excellent oral drug competency with the ability for gastrointestinal absorption and blood-brain-barrier permeation with nil toxicity.Molecular docking depicted thujone with the strongest affinity for mammalian target of rapamycin,phosphoinositide 3-kinase,and protein kinase B followed by B-cell lymphoma 2,murine double minute 2 and adenosine monophosphate-activated protein kinase,which might act as upstream triggers of apoptotic-autophagic crosstalk.Conclusion:Robust"multi-targeting"anticancer potential of Thuja drug and thujone for HPV-infected cervical cancer ascertained its therapeutic efficacy for HPV infections.

Asmita Pal;Sucharita Das;Soumalee Basu;Rita Kundu

Department of Botany,University of Calcutta,Kolkata,West Bengal 700019,India;Department of Microbiology,University of Calcutta,Kolkata,West Bengal 700019,India

结合医学学报（英文版）

[1]Asmita Pal;Sucharita Das;Soumalee Basu;Rita Kundu-.Apoptotic and autophagic death union by Thuja occidentalis homeopathic drug in cervical cancer cells with thujone as the bioactive principle)[J].结合医学学报（英文版）,2022(05):463-472

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