Effect of LY294002 on expression of anti-/pro- inflammatory factors and ROS generation.

<p>(A-C) mRNA expression of IL-6, VCAM-1 and eNOS after LSS was applied with or without LY294002 pre-treatment compared with ivabradine pre-treated cells. (D-F) Bar diagram showing quantitative data of immunofluorescence positive cells. Cells in bright green were counted for five different visual fields. (G-I) Immunofluorescence of IL-6, VCAM-1 and eNOS-Thr495 generated by LSS. (J-K) LY294002 was applied before ivabradine pre-treatment and LSS exposure, ROS generation was compared within groups. (L-M) Bar diagram showing quantitative data of ROS-positive cells. *p<0.05 versus LSS 0 minute, # p<0.05 versus LSS 30 minutes, $p<0.05 versus Iva 30 minutes.</p

Effect of ivabradine on LSS induced changes of eNOS and mTOR pathway.

<p>(A) ECs were exposed to LSS for 30 minutes after pre-treated with ivabradine, protein phosphorylation was compared in cells exposed to LSS with or without pre-treatment of ivabradine. (B-G) Bar diagram showing intensity data of western blot images, all data were normalized by GAPDH. (H) mTORC2 pathway expression in cells exposed to LSS for 120 minutes after pre-treated with ivabradine. (I-K) Bar diagram showing intensity data of western blot images, all data were normalized by GAPDH. *p<0.05 versus LSS 0 minute, # p<0.05 versus LSS 30 or 120 minutes.</p

Immunofluorescence of IL-6, VCAM-1 and eNOS-Thr495 generated by LSS.

<p>(A-F) Representative images of immunofluorescence. Positive cells in bright green were counted for five different visual fields. (G-L) Bar diagram showing quantitative data of positive cells. *p<0.05 versus LSS 0 minute, # p<0.05 versus LSS 30 or 120 minutes.</p

Fluorescence microscopy of ROS-positive cells stained with DHE and DCF and nucleus stained with DAPI.

<p>(A, E) DHE-positive cells in red were counted for five different visual fields. (B, F) DCF-positive cells in bright green were counted for five different visual fields. (C, G) Bar diagram showing quantitative data of DHE-positive cells. (D, H) Bar diagram showing quantitative data of DCF-positive cells. *p<0.05 versus LSS 0 minute, # p<0.05 versus LSS 30 or 120 minutes.</p

Effect of LSS on eNOS and mTOR pathway.

<p>(A) Representative western blot images of eNOS-Thr495, p70S6K-Thr389, S6RP-Ser235/236, raptor-Ser792, rictor-Thr1135, Akt-Ser473 and GAPDH that exposed to LSS for time periods. (B-G) Line charts showing intensity of western blot images, all data were normalized by GAPDH. *p<0.05 versus LSS 0 minute.</p

Effect of LY294002 on eNOS and mTOR pathway.

<p>(A) LY294002 was applied before ivabradine pre-treatment and LSS exposure, protein phosphorylation was compared in cells exposed to LSS for 30 minutes with or without ivabradine pre-treatment. (B-F) Bar diagram showing intensity data of western blot images, all data were normalized by GAPDH. *p<0.05 versus LSS 0 minute, # p<0.05 versus LSS 30 minutes, $p<0.05 versus Iva 30 minutes.</p

LSS induced changes of anti-/pro- inflammatory factors.

<p>(A) mRNA expression of IL-6, VCAM-1 and eNOS after LSS was applied for 0, 30 and 120 minutes. (B-G) Ivabradine reverses LSS caused mRNA alteration of IL-6, VCAM-1 and eNOS. All experiments above were repeated three times.</p

Anticancer Platinum(IV) Prodrugs Containing Monoamino­phosphonate Ester as a Targeting Group Inhibit Matrix Metalloproteinases and Reverse Multidrug Resistance

A novel class of platinum­(IV) complexes comprising a monoamino­phosphonate ester moiety, which can not only act as a bone-targeting group but also inhibit matrix metalloproteinases (MMPs), were designed and synthesized. Biological assay of these compounds showed that they had potent antitumor activities against the tested cancer cell lines compared with cisplatin and oxaliplatin and indicated low cytotoxicity to human normal liver cells. Particularly, the platinum­(IV) complexes were very sensitive to cisplatin resistant cancer cell lines. The corresponding structure–activity relationships were studied and discussed. Related mechanism study revealed that the typical complex <b>11</b> caused cell cycle arrest at S phase and induced apoptosis in Bel-7404 cells via a mitochondrial-dependent apoptosis pathway. Moreover, complex <b>11</b> had potent ability to inhibit the tumor growth in the NCI-H460 xenograft model comparable to cisplatin

Design, Synthesis, and Biological Features of Platinum(II) Complexes with Rigid Steric Hindrance

A series of platinum­(II) complexes, with N-monosubstituted 1<i>R</i>,2<i>R</i>-diaminocyclohexane bearing methoxy-substituted benzyl groups as carrier ligands, were designed and synthesized. The newly prepared compounds, with chloride anions as leaving groups, were found to be very active against the tested cancer cell lines, including a cisplatin-resistant cell line. Despite their efficacy against tumor cells, they also showed low toxicity to a human normal liver cell line. Among them, complex <b>1</b> had superior cytotoxic activity against A549, HCT-116, MCF-7, SGC7901, and SGC7901/CDDP cancer cell lines. The DNA binding assay is of further special interest, as an unusual monofunctional binding mode was found, due to the introduction of a rigid substituted aromatic ring in the 1<i>R</i>,2<i>R</i>-diaminocyclohexane framework as steric hindrance. The linkage of complex <b>1</b> with DNA was stable and insensitive to nucleophilic attack. Moreover, studies including cellular uptake, gel electrophoresis, apoptosis and cell cycle, and Western blot analysis have provided insight into the high potency of this compound

Combretastatin A‑4 Analogue: A Dual-Targeting and Tubulin Inhibitor Containing Antitumor Pt(IV) Moiety with a Unique Mode of Action

Three new Pt­(IV) complexes comprising a combretastatin A-4 analogue were designed and synthesized. The resulting antitumor Pt­(IV) complexes could significantly improve the antiproliferative activity and overcome the drug resistance of cisplatin in vitro. Interestingly, these novel compounds not only can carry the DNA binding Pt­(II) warhead into the cancer cells but also have a small molecule fragment that can inhibit tubulin polymerization. Among them, complex <b>13</b>, which was attached to an inhibitor of tubulin at one axial position of Pt­(IV) octahedral coordination sphere, could effectively enter cancer cells, arrest the cell cycle in HepG-2 cancer cells at G2/M phases, and induce activation of caspases triggering apoptotic signaling via the mitochondrial-dependent apoptosis pathways. Moreover, complex <b>13</b> has the ability to effectively inhibit the tumor growth in the HepG-2 xenograft model without causing significant loss of animal body weight in comparison with cisplatin