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It is known that autophagy-deficient cells are prone to DNA damage, but the specific role of autophagy in DNA damage repair is not fully known. Here, we show that autophagy-deficient liver cancer cells exhibit increased DNA damage caused by the chemotherapeutic agent epirubicin. Autophagy deficiency promotes downregulation of the DNA repair enzyme O6methylguanine-DNA methyltransferase (MGMT) in liver cancer cells. However, autophagy induction with epirubicin had no impact on MGMT gene or protein expression in liver cancer cells. In the absence of autophagy, the chemosensitivity of liver cancer cells was increased, but this was reversed by MGMT overexpression, indicating that autophagy mediates resistance to chemotherapy in liver cancer cells via MGMT. These findings demonstrate a direct link between autophagy, MGMT, and DNA damage repair in liver cancer cells, and show that MGMT not only regulates chemosensitivity to alkylating agents, but may also be involved in other DNA damage repair processes in autophagy-deficient cells.

Capsaicin, a functional component of chili pepper, possesses anti-inflammatory, analgesic, and anti-cancer properties. This study aimed to determine the property of capsaicin against hepatocarcinogenesis in vivo and investigate the role of the SIRT1/SOX2 pathway in the mode of action of capsaicin in hepatic progenitor cells (HPCs), which is related to hepatocarcinogenesis.We prepared a diethylnitrosamine-induced liver cancer model in rats to examine hepatocarcinogenesis, and delivered liposomal capsaicin through the subcutaneous transposition of the spleen to the liver. Liver sections from rats and hepatocarcinoma patients were stained for the markers of HPCs or SIRT1/SOX2 signaling. SIRT1/SOX2 signalling expression was measured using immunoprecipitation and western blot.We found that capsaicin significantly inhibited hepatocarcinogenesis. Notably, capsaicin inhibited HPCs activation in vivo but did not induce apoptosis in the normal hepatic progenitor cell line in rats in vitro. This suggests that capsaicin suppresses hepatocarcinogenesis by inhibiting the stemness of HPCs. Moreover, capsaicin can induce this inhibition by reducing the stability of SOX2. SIRT1 is overexpressed in liver cancer and acts as a tumor promoter via SOX2 deacetylation. Using immunoprecipitation, we identified direct binding between SIRT1 and SOX2. The capsaicin treatment resulted in SIRT1 downregulation which reduced deacetylation, and increased nuclear export as well as subsequent ubiquitous degradation of SOX2.Altogether, we report that capsaicin suppresses hepatocarcinogenesis by inhibiting the stemness of HPCs via SIRT1/SOX2 signaling. It may serve as a promising therapeutic candidate for liver cancer.

Hepatocellular carcinoma (HCC) generally occurs in the presence of chronic liver injury, often as a sequela of liver fibrosis. Hepatic progenitor cells (HPCs) are known to be capable of forming both hepatocytes and cholangiocytes in chronic liver injury, which are also considered a source of myofibroblasts and tumor-initiating cells, under carcinogenic circumstances. However, the underlying mechanisms that activate HPCs to give rise to HCC are still unclear. In current study, the correlation between HPCs activation and liver fibrosis and carcinogenesis was investigated in rats and human specimens. We analyzed the role of HPCs in tumorigenesis, by transplanting exogenous HPCs in a diethylnitrosamine-induced rat HCC model. Our data indicated that HPC activation correlated with hepatic fibrosis and hepatocarcinogenesis. We further found that exogenous HPC infusion promoted liver fibrosis and hepatocarcinogenesis, while lipopolysaccharides (LPS) played an important role in this process. However, results of our study indicated that LPS did not induce HPCs to form tumor in nude mice directly. Rather, LPS induced myofibroblast-like morphology in HPCs, which enhanced the tumorigenic potential of HPCs. Further experiments showed that LPS/Toll-like receptor 4 (TLR4) signaling mediated the differentiation of HPCs into myofibroblasts and enhanced the production of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), which led to the aberrant expression of Ras and p53 signaling pathways in HPCs, and finally, promoted the proliferation and malignant transformation of HPCs, by long non-coding RNA regulation. Besides, examination of HCC clinical samples demonstrated that IL-6 and TNF-α production correlated with HPC activation, hepatic fibrosis, and HCC recurrence. Our study indicates that both myofibroblasts and tumor cells are derived from HPCs. HPC-derived myofibroblasts create tumor microenvironment and contribute to the proliferation and malignant transformation of HPCs. Furthermore, LPS present in the chronic liver inflammation microenvironment might play an important role in hepatocarcinogenesis, by regulating the plastic potential of HPCs.

Hypoxia is a common feature of most solid tumors and its precise imaging is of great importance to therapy planning. Here we demonstrated that a ruthenium(II) anthraquinone complex [Ru(bpy)2(ipad)](ClO4)2 (Ru-ipad, where bpy = 2,2′-bipyridine and ipad = 2-(anthracene-9,10-dione-2-yl)imidazo[4,5-f][1,10]phenanthroline), is a powerful optical probe that can detect repeated cycles of hypoxia–normoxia in living cells in real time. This hypoxia probe also showed cytotoxicity against hypoxic 2D cancer monolayer cultures and 3D multicellular tumor spheroids (MCTSs). The cytotoxicity of Ru-ipad was in accordance with its suppression of the hypoxia-inducible factor-1α (HIF-1α) protein in tumor cells. Therefore, Ru-ipad not only has the potential to detect hypoxia with high sensitivity but also displays hypoxia-targeted antitumor effects through the inhibition of HIF-1α expression. The discovery of this dual-function ruthenium(II) anthraquinone complex may represent an important advance in hypoxic solid tumor treatment.

A cyclometalated iridium(III) complex, [Ir(ppy)2(PCN)]Cl (Ir1, ppy = 2-phenylpyridine, PCN = 2-(4-cyanophenyl)imidazo[4,5-f] [1,10] phenanthroline), was synthesized and characterized in the present study. Ir1 inhibited the proliferation, migration and invasion of MDA-MB-231 human breast cancer cells in a dose-dependent manner. Moreover, Ir1 down-regulated the phosphorylation of AKT/ERK signal pathways. According to confocal fluorescence microscopy analysis, Ir1 was primarily localized within the mitochondria and induced apoptosis through an intrinsic mitochondria-mediated apoptotic pathway. Thus, Ir1 exhibited both antimetastatic and antineoplastic properties, indicating that Ir1 may be a viable drug candidate in antimetastasis and anticancer therapies.

A new cyclometalated iridium(III) complex [Ir(2-pq)2(HPIP)]Cl (Ir1, 2-pq=3-phenylisoquinoline, HPIP=2-(2-hydroxyphenyl)imidazo[4,5-f]1,10-phenanthroline) was synthesized and applied to image mitochondria in living cells. Ir1 displayed excellent ability to selectively accumulate in mitochondria of live cells with no requirement of replacement of the culture medium. Due to Ir1 exhibiting better photostability than the commercially available mitochondrial trackers, this complex could be applicable for the imaging and tracking of the mitochondrial morphological changes over long periods of time. In addition, in vitro cytotoxicity investigation revealed that Ir1 showed negligible cytotoxicity at the concentrations employed. Based on these, Ir1 is suitable for organelle-selective imaging in living cells.

Strategies for targeting CD47 are becoming a hot spot of cancer immunotherapy. However the ubiquitous expression of CD47, especially on the RBC, makes the targeted therapy facing safety risk issues. So, how to balance the safety and efficacy during CD47 inhibition is currently a major question. We had reported an anti-CD47 antibody ZF1 with potent anti-tumor effect. In this study, we further developed and assessed a novel fully human anti-CD47 antibody, AMMS4-G4, derived from ZF1 using affinity maturation. AMMS4-G4 exhibited equivalent anticancer effects with Hu5F9-G4, a humanized anti-CD47 antibody in clinical trial, on the potential of inducing significant phagocytosis of tumor cells in vitro and prolonging the survival of leukemia xenografted mice. Additionally, AMMS4-G4 significantly inhibited the growth of grafted solid tumors by enhancing macrophage infiltration and modestly enhanced the anti-tumor activity of opsonizing antibody and antiangiogenic therapy. In cynomolgus monkeys, AMMS4-G4 was safely administered, was well tolerated at doses of 30 and 60 mg/kg, and did not produce serious adverse events, except for the reversible anemia, which was observed after 3 days and started to recover from 9 days later. Remarkably, it was proved by in vitro assay that Hu5F9-G4 induced RBC hemagglutination which wasn't observed in AMMS4-G4. On the whole, AMMS4-G4 was demonstrated to be a promising candidate with great potential and safe profile for cancer immunotherapy.

A novel cyclometalated iridium(III) complex, [Ir(CˆN)2(HPIP)]Cl (IrC) was synthesized and characterized. IrC exhibited about 10-fold higher cytotoxicity than cisplatin against A549 cancer cell line. Interestingly, a cisplatin-resistant cell line, A549-CP/R showed sensitivity to IrC. Further study suggested that IrC induced apoptosis via negatively regulated the nuclear factor-kappa B (NF-κB) pathway, which involved reactive oxygen species (ROS) generation, Bcl-2 and caspase family members activation. Taken together, these results suggest that IrC is able to overcome chemoresistance and may be an effective treatment for platinum-resistant cancer therapy.

Hepatocellular carcinoma (HCC) is a typical inflammation-related cancer. Hepatitis B virus (HBV) and hepatitis C virus (HCV) infection are well-known leading causes of HCC. However, the mechanism of the induction of HCC by these virus is still being debated. This review will focus on the current knowledge of the pathogenesis of HBV- and HCV-induced inflammation and the role of such immune activation in the tumorigenesis of HCC. It is well established that the recruitment of certain number and type of immune cells to liver is essential for the resolution of HBV and HCV infection and the prevention of subsequent chronic persistent infection. However, in case that the immune response do not completely clear virus, persistent chronic infection occurs, and the perpetual immune response may contribute to chronic damages of the liver. Such chronic inflammatory damages further harm hepatocytes, but not hepatic progenitor cells (HPCs). Thus, following chronic damages, HPCs are activated and their dysregulated proliferation ensures survival in the hostile environment, contributing to the tumorigenesis of HCC. Furthermore, accumulating evidence also provides a strong link between HPCs and human hepatocellular carcinoma. Collectively, these findings support a notion that immune response is involved in liver damage during hepatitis virus infection, and the activation and dysregulated differentiation of hepatic progenitor cells promote the tumorigenesis of human hepatocellular carcinoma.

Autophagy plays important roles in self-renewal and differentiation of stem cells. Hepatic progenitor cells (HPCs) are thought to have the ability of self-renewal as well as possess a bipotential capacity, which allows them to differentiate into both hepatocytes and bile ductular cells. However, how autophagy contributes to self-renewal and differentiation of hepatic progenitor cells is not well understood. In this study, we use a well-established rat hepatic progenitor cell lines called WB-F344, which is treated with 3.75 mM sodium butyrate (SB) to promote the differentiation of WB-F344 along the biliary phenotype. We found that autophagy was decreased in the early stage of biliary differentiation, and maintained a low level at the late stage. Activation of autophagy by rapamycin or starvation suppressed the biliary differentiation of WB-F344. Further study reported that autophagy inhibited Notch1 signaling pathway, which contributed to biliary differentiation and morphogenesis. In conclusions, autophagy regulates biliary differentiation of hepatic progenitor cells through Notch1 signaling pathway.

Impacts of 33-yr of application of S-containing and Cl-containing chemical fertilizers on rice (Oryza sativa L.) yield and its components were investigated in a red paddy field experiment, south China. The treatments included: 1) adding 302 kg SO42−-S ha−1 yr−1 with application of (NH4)2SO4, K2SO4, and calcium superphosphate (SO42−); 2) adding 56 kg SO42−-S and 176 kg Cl ha−1 yr−1 with application of urea, calcium superphosphate, and KCl (Cl−+SO42−); 3) adding 516 kg Cl ha−1 yr−1 with application of NH4Cl, KCl, and KH2PO4 (Cl−). Under each treatment, the applied N, P, and K nutrients were controlled at conventional rates of 150 kg N ha−1 yr−1, 75 kg P2O5 ha−1 yr−1, 225 kg K2O ha−1 yr−1, respectively. Under the S-containing fertilizer application, soil SO42−-S content showed a first increasing then decreasing trend with years, and was significantly negatively correlated with annual rice yield. Average annual yield significantly declined in an order of Cl−, Cl−+SO42−, and SO42−. Under the Cl− treatment, soil SO42−-S content was maintained at about 26.5 mg kg−1, not showing deficiency. From 1990 to 2000, rice yield declined rapidly under the SO42− treatment, and was significantly lower than that under the Cl− treatment. After then, there was no significant difference in yield among the treatments. Our results demonstrated that long-term application of S-containing fertilizer could result in excessive accumulation of SO42−-S in the red paddy soils of south China, therefore producing a certain threat to rice growth. The Cl-containing fertilizer could be relatively safe.

In recent years, the data mining technology has been developed rapidly. New efficient algorithms are emerging. Association data mining plays an important role in data mining, and the frequent item sets are the highest and the most costly. This paper is based on the association rules data mining technology. The advantages and disadvantages of Apriori algorithm and FP-growth algorithm are deeply analyzed in the association rules, and a new algorithm is proposed, finally, the performance of the algorithm is compared with the experimental results. It provides a reference for the extension and improvement of the algorithm of association rule mining.

A series of novel chiral ruthenium(II) polypyridyl complexes (Δ-Ru1, Λ-Ru1, Δ-Ru2, Λ-Ru2, Δ-Ru3, Λ-Ru3) were synthesized and evaluated to determine their antiproliferative activities. Colocalization, inductively coupled plasma mass spectrometry, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay studies showed that these ruthenium(II) complexes accumulated preferentially in the mitochondria and exhibited cytotoxicity against various cancer cells in vitro. The complex Δ-Ru1 is of particular interest because it was found to have half-maximal inhibitory concentrations comparable to those of cisplatin and better activity than cisplatin against a cisplatin-resistant cell line, A549-CP/R. Δ-Ru1 induced alterations in the mitochondrial membrane potential and triggered intrinsic mitochondria-mediated apoptosis in HeLa cells, which involved the regulation of Bcl-2 family members and the activation of caspases. Taken together, these data suggest that Δ-Ru1 may be a novel mitochondria-targeting anticancer agent.

Three mononuclear copper complexes [Cu(PDTP)Cl2] (PDTP = 4-phenyl-2,6-di(thiazole-2-yl)pyridine, CuPDTP), [Cu(ADTP)Cl2] (ADTP = 4-(anthracen-9-yl)-2,6-di(thiazole-2-yl)pyridine, CuADTP) and [Cu(BFDTP)Cl2] (BFDTP = 4-(benzofuran-2-yl)-2,6-di(thiazole-2-yl)pyridine, CuBFDTP) were synthesized and characterized. The X-ray single crystallography results indicated that the Cu(II) ions showed slightly distorted square pyramid coordination environments, and the ligands deviated from ideal planarity in all three compounds. Based on the DNA binding studies, it was demonstrated that these three complexes exhibited weak DNA binding strengths, which were most likely groove binding modes. CuPDTP, CuADTP and CuBFDTP induced efficient DNA cleavage in the dark without the addition of external catalysts (oxidant or reductant). In contrast, in the presence of reducing or oxidizing agents, the nuclease activities increased more than 10-fold. Mechanistic investigations revealed the participation of reactive oxygen species, which can be trapped by ROS radical scavengers and ROS sensors. In the same experimental conditions, the free ligands and CuCl2 did not display any DNA cleaving activity. This result indicates that the complexes, rather than their components, play a significant role in the nuclease reaction process and that DNA cleavage may be initiated in an oxidative pattern. The proposed mechanism was attributed to the in situ activation of molecular oxygen by the oxidation of the copper complexes. In the MTT cytotoxicity studies, the three Cu(II) complexes exhibited an antitumor activity against the HeLa, BEL-7402 and HepG2 tumor cell lines. The HeLa cells treated with Cu(II) complexes demonstrated marked changes in their nuclear morphology, which were detected by Hoechst 33258 nuclear staining and acridine orange/ethidium bromide (AO/EB) staining assays. Nuclear chromatin cleavage also was observed from alkaline single-cell gel electrophoresis (comet assay).

A series of chiral Ru(II) complexes bearing thiophene ligands were synthesized and characterized. Both Ru(II) complexes Δ/Λ-[Ru(bpy)2(pscl)](2+) (Δ/Λ-1) and Δ/Λ-[Ru(bpy)2(psbr)](2+) (Δ/Λ-2) (bpy=2,2'-bipyridine, pscl=2-(5-chlorothiophen-2-yl)imidazo[4,5-f][1,10]phenanthroline, psbr=2-(5-bromothiophen-2-yl)imidazo[4,5-f][1,10]phenanthroline) showed antitumor activities against A549, HepG2 and BEL-7402 tumor cell lines, especially HeLa tumor cell line. Moreover, Δ enantiomers were more active than Λ enantiomers, accounting for the different cellular uptake. In addition, with the extension of time, these enantiomers could finally accumulate in the nucleus, suggesting that nucleic acids were the cellular target of these enantiomers. The DNA-binding behaviors of complexes were studied using spectroscopic and viscosity measurements. Results suggested that four complexes could bind to DNA in an intercalative mode but no obvious DNA-binding selectivity between the enantiomers was observed. Topoisomerase inhibition and DNA religation assay confirmed that four complexes acted as efficient dual topoisomerase I and II poisons, DNA strand breaks had also been observed from alkaline single cell gel electrophoresis (comet assay). Δ-1 and Δ-2 inhibited the growth of HeLa cells through the induction of apoptotic cell death, as evidenced by the Alexa Fluor® 488 annexin V staining assays and flow cytometry analysis. The results demonstrated that Δ/Λ-1 and Δ/Λ-2 acted as dual topoisomerase I and II poisons and caused DNA damage that could lead to cell cycle arrest by apoptosis.

Three new tridentate copper(II) complexes [Cu(dthp)Cl(2)] (1) (dthp=2,6-di(thiazol-2-yl)pyridine), [Cu(dmtp)Cl(2)] (2) (dmtp=2,6-di(5-methyl-4H-1,2,4-triazol-3-yl)pyridine) and [Cu(dtp)Cl(2)] (3) (dtp=2,6-di(4H-1,2,4-triazol-3-yl)pyridine) have been synthesized and characterized. Crystal structure of complex 1 shows that the complex existed as distorted square pyramid with five co-ordination sites occupied by the tridentate ligand and the two chlorine anions. Ethidium bromide displacement assay, viscosity measurements, circular dichroism studies and cyclic voltammetric experiments suggested that these complexes bound to DNA via an intercalative mode. Three Cu(II) complexes were found to efficiently cleave DNA in the presence of sodium ascorbate, and singlet oxygen ((1)O(2)) and hydrogen peroxide were proved to contribute to the DNA cleavage process. They exhibited anticancer activity against HeLa, Hep-G2 and BEL-7402 cell lines. Nuclear chromatin cleavage has also been observed with AO/EB staining assay and the alkaline single-cell gel electrophoresis (comet assay). The results demonstrated that three Cu(II) complexes cause DNA damage that can induce the apoptosis of BEL-7402 cells.