Targeting Aberrant Expression of STAT3 and AP-1 Oncogenic Transcription Factors and HPV Oncoproteins in Cervical Cancer by Berberis aquifolium

Abstract

Background: Present study examines phytochemical preparation that uses berberine’s plant source B. aquifolium root for availability of similar anti-cervical cancer (CaCx) and anti-HPV activities to facilitate repurposing of the B. aquifolium based drug in the treatment of CaCx.Purpose: To evaluate therapeutic potential of different concentrations of ethanolic extract of B. aquifolium root mother tincture (BAMT) against HPV-positive (HPV16: SiHa, HPV18: HeLa) and HPV-negative (C33a) CaCx cell lines at molecular oncogenic level.Materials and Methods: BAMT was screened for anti-proliferative activity by MTT assay. Cell cycle progression was analyzed by flowcytometry. Then, the expression level of STAT3, AP-1, HPV E6 and E7 was detected by immunoblotting, whereas nuclear localization was observed by fluorescence microscopy. Phytochemicals reportedly available in BAMT were examined for their inhibitory action on HPV16 E6 by in silico molecular docking.Results: BAMT induced a dose-dependent decline in CaCx cell viability in all cell types tested. Flowcytometric evaluation of BAMT-treated cells showed a small but specific cell growth arrest in G1-phase. BAMT-treatment resulted in reduced protein expression of key transcription factors, STAT3 with a decline of its active form pSTAT3 (Y705); and components of AP-1 complex, JunB and c-Jun. Immunocytochemistry revealed that BAMT did not prevent the entry of remnant active transcription factor to the nucleus, but loss of overall transcription factor activity resulted in reduced availability of transcription factors in the cancer cells. These changes were accompanied by gradual loss of HPV E6 and E7 protein in BAMT-treated HPV-positive cells. Molecular docking of reported active phytochemicals in B. aquifolium root was performed, which indicated a potential interference of HPV16 E6’s interaction with pivotal cellular targets p53, E6AP or both by constituent phytochemicals. Among these, berberine, palmatine and magnoflorine showed highest E6 inhibitory potential.Conclusion: Overall, BAMT showed multi-pronged therapeutic potential against HPV infection and cervical cancer and the study described the underlying molecular mechanism of its action.

SOCS5 contributes to temozolomide resistance in glioblastoma by regulating Bcl-2-mediated autophagy

“…For the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay used to detect the cell proliferation rate [ 30 ], transfected/treated cells were seeded in 96-well cell culture plates (4 × 10 3 cells/well). Cells were allowed to adhere for 12 h and then treated with chemotherapeutic agents (final concentration, 40 μM).…”

Evaluation of Therapeutic Potential of Selected Plant-Derived Homeopathic Medicines for their Action against Cervical Cancer

“…The MTT cell viability assay was performed as described earlier 8 9 with minor modifications. Briefly, CaCx cells (2 × 10 3 /w) were seeded in a 96-well plate in a final volume of 0.1 mL of culture medium DMEM (SiHa and HeLa)/MEM (C33a) supplemented with 10% FBS.…”

“…In silico modelling of molecular docking of phytochemicals with HPV16 E6 was performed following the method described earlier. 8 Briefly, two crystallographic protein structures of HPV16 E6 were used, i.e. chain B of 6SJA (aa7-158; resolution 1.5 Å), which represents the native conformation of E6 (non-p53 interacting) 14 ; and chain F of 4XR8 (aa8-158; resolution 2.25 Å), which represents the p53-interacting conformation of E6 15 .…”

“…There are several studies demonstrating potential anti-HPV therapeutics of plant origin 7 8 . Pure phytochemicals (curcumin and berberine 9 ), crude extracts ( Bryophyllum pinnata leaf extract 10 and Phyllanthus emblica fruit pulp extract 11 ) or rationally-designed combinations of plant derivatives (Praneem 12 and Basant 13 ) showed promising anti-HPV and anti-CaCx results in both pre-clinical studies (curcumin, berberine, B. pinnata, and P. emblica ) and placebo-controlled, double-blind clinical trials (Praneem, curcumin, and Basant).…”

Synthesis of benzothiazole-appended bis-triazole-based structural isomers with promising antifungal activity against Rhizoctonia solani

“…To observe the putative binding mode of the compounds to the targeted protein, sterol 14a-demethylase enzyme (CYP51), (PDB entry: 3GW9) molecular docking studies have been performed with minor modications by using AutoDock Vina. 24,25 Deep analysis of the 2D and 3D poses of compounds 5f, 6, 7f, and 8 were performed in a similar manner to that of hexaconazole, the reference drug (Fig. 5-9).…”