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The present study aims in vitro cell suspension culture of Bridelia retusa, isolation of anthocyanin, purification, fractionation and its anti-metastatic potential against oral squamous carcinoma cells. Experimental results reveal that 2, 4-D either alone or in combination with kinetin supplemented in MS medium showed significant initiation of callus from leaf explants than stem. Growth hormones, pH, light, and carbon source influence anthocyanin synthesis. Maximum callus induction was noticed with 2.5 mg/L N6-benzyladenine (BA) + 2 mg/L 2, 4-dichlorophenoxyacetic acid (2, 4-D) (98.9%). Fresh and dry weight of the calli were i.e., 1.9 ± 0.04 and 0.45 ± 0. 03 g respectively. Optimal response was seen with light on MS medium contain 4% glucose + 2.5 mg/L BA and 2 mg/L 2, 4-D at pH 3.5 yielded 2.8 mg /g of anthocyanins. Suspension culture medium fortified with 2, 4-D (2.5 mg/L) + BA (2 mg/L) at pH 5.0 induced anthocyanin production at pH 4.4 - 4.6. HCl-ethanol extraction for 90 min yielded the maximum anthocyanin content. Fractionation of anthocyanin using HPLC coupled with mass spectrometry revealed 07 fractions such as acylated cyanidins, two peonidins, cyanidin 3-p-coumaroyl and feruloyl diglucoside-5-glucosides. In the search of novel therapeutic drugs against cancer, cytotoxicity effect of B.retusa anthocyanin extracts on human oral squamous cell carcinoma (SCC4, SCC9 and SCC25) cells using cell adhesion and cell viability assay was carried. The morphological alterations in SCCs cells after treatment with B.retusa anthocyanin includes nuclear condensation, fragmentation and apoptotic cells as revealed by Hoechst stain. Flow cytometry showed arresting of SCC25 cells mostly in the G0/G1 and S-G2/M stages with a concomitant up regulation of sub-G1 fraction, indicating cell death by apoptosis. Apoptosis was further substantiated by the activation of caspase-3 expression in the SCC25 cells treated with B.retusa anthocyanin. Thus, it is possible to suggest that B.retusa anthocyanin cause apoptosis of SCCs and warrant further investigation using animal models. [ABSTRACT FROM AUTHOR]

Background: Activation of the Wnt pathway is known to promote tumorigenesis and tumor metastasis, and targeting Wnt pathway inhibition has emerged as an attractive approach for controlling tumor invasion and metastasis. The major pathway for inhibiting Wnt is through the degradation of β-catenin by the GSK3-beta/CK1/Axin/APC complex. It was found that Hep3B hepato-carcinoma cells respond to anthocyanins through GSK3-beta-induced suppression of beta-catenin; however, they cannot dephosphorylate GSK3-beta without AMPK activation. Methods: We tested the effects of anthocyanins on proliferation and apoptosis by MTT and Annexin V-PI staining in vitro. Mouse xenograft models of hepato-carcinomas were established by inoculation with Hep3B cells, and mice were injected with 50 mg/kg/ml of anthocyanins. In addition, protein levels of p-GSK3-beta, beta-catenin, p-AMPK, MMP-9, VEGF, and Ang-1 were also analyzed using western blot. Results: Anthocyanins decrease phospho-GSK3-beta and beta-catenin expression in an in vivo tumor xenograft model, increase AMPK activity in this model, and inhibit cell migration and invasion, possibly by inhibiting MMP-2 (in vitro) and the panendothelial marker, CD31 (in vivo). To elucidate the role of the GSK3-beta/beta-catenin pathway in cancer control, we conditionally inactivated this pathway, using activated AMPK for inhibition. Further, we showed that AMPK siRNA treatment abrogated the ability of anthocyanins to control cell proliferation and metastatic potential, and Compound C, an AMPK inhibitor, could not restore GSK3-beta regulation, as exhibited by anthocyanins in Hep3B cells. Conclusion: These observations imply that the AMPK-mediated GSK3-beta/beta-catenin circuit plays crucial roles in inhibiting cancer cell proliferation and metastasis in anthocyanin-treated hepato-carcinoma cells of Meoru origin. [ABSTRACT FROM AUTHOR]

• Synthesis and characterization of benzimidazolyl‑hydroxy-coumarinyl diad. • Spectroscopic and Single Crystal X-ray structure determination of the diad. • Diad is nonfluorescent and binding with Al3+ in presence of fourteen other cations shows green emission. • The anti-metastatic activity of compound on Human skin carcinoma (A431) and Human lung carcinoma (A549) cells are examined. 8-(1H-Benzoimidazol-2-yl)-7‑hydroxy-4-methyl-chromen-2-one (HL), a benzimidazolyl‑hydroxy-coumarinyl diad, serves as fluorescent turn-on Al3+-sensor in acetonitrile-water (2:1, v/v) of pH, 7.2 (HEPES buffer) medium. The intensity of emission on binding with Al3+ is enhanced by twelve times of probe with limit of detection (LOD) 0.62 µM. The structure of the probe has been established by the spectroscopic data (FT-IR, 1H NMR, Mass spectra) and is confirmed by Single Crystal X-ray structure determination. The co-ordination environment of the complex, [Al(L)(NO 3) 2 (H 2 O)(NCCH 3)], has been supported by Mass, FT-IR, NMR spectra and the Job's plot supports 1:1 composition. The anti-metastatic potential of HL on Human skin carcinoma (A431) and Human lung carcinoma (A549) cells are examined. Wound healing and Zymography assay suggest that HL has anti-migratory properties. The RT-PCR and Immunoblotting data suggest that HL down-regulates the expression of mesenchymal markers (Vimentin, Snail, Slug, MMP, and MMP9) and induces the expression of the epithelial marker (E-Cadherin). [Display omitted] [ABSTRACT FROM AUTHOR]

Studies have shown that metastases are the life-threatening factor in prostate cancer, as in other cancers. Especially, upregulation of voltage-gated Na+ channels (VGSCs) in the cell membrane plays a role in the progression of primary tumor to metastasis. VGSCs with increased expression are responsible for migration and invasion, which are the main steps of metastasis. It has been determined as in vitro and in vivo that some ion channel blockers, which block upregulated VGSCs and are used in the treatment of different diseases, suppress the movement of cancer cells and inhibit metastasis. Inhibiting the movement of cancer cells with these drugs that show pharmacological effects by suppressing VGSCs will open a new field of treatment for anti-metastatic potential. Ranolazine (RNL), which is clinically used in the treatment of angina pectoris, has a therapeutic effect on heart pain, contraction and dysfunction by blocking sodium channels. On the other hand, Riluzole (RIL) is a neuroprotective drug, especially used in the ALS disease, which inhibits sodium and calcium flow and plays a role in the treatment by suppressing glutamate release. These VGSC blockers have higher activity in the hypoxic environment than the normoxic environment. The aim of this study is to determine whether the combined treatment using two VGSC blockers together had a synergistic effect on the migration of Dunning model high metastatic prostate cancer cells, in other words, to determine their contribution to antimetastatic potential and to search the potential for developing a new treatment protocol against prostate cancer metastasis. The effects of RNL (2.5 μM, 5 μM), RIL (2.5 μM, 5 μM) and RNL + RIL (2.5+2.5 μM, 5+2.5 μM) on the proliferation of Dunning model rat prostate cancer MATLyLu and AT-2 cells with different metastatic features were examined. The effects on the anti-metastatic potential were realized by "wound heal" method in high metastatic Mat-LyLu cells. According to the results obtained, when both RNL and RIL were administered alone, there was no significant change in the proliferation of the cells. Combined administration of drugs (5+5 μM) to MAT-LyLu cells reduced cell proliferation by 30%. TTX which is a VGSC blocker, blocked the lateral movement of MaTLyLu cells by 23%. RNL and RIL which did not affect cell proliferation, produced inhibition of cell migration (48h) both alone and in combination at varying rates (5-14%). The combination of RNL and RIL which was searched for the first time, produced inhibition of migration of prostate cancer cells under normoxic conditions, but did not produce a synergistic effect. These promising results suggest that the combined effects of RNL and RIL on anti-metastatic potential should be searched in detail in hypoxic conditions and in vivo experimental models. [ABSTRACT FROM AUTHOR]