Establishment of malignant pleural effusion model of Lewis lung cancer based on Micro Echo Technology / 中国生化药物杂志

Objective To establish a malignant pleural effusion model of Lewis lung cancer in C57BL/6 mice based on Micro Echo technology. Methods Single tumor cell suspension of Lewis lung cancer was injected into thoracic cavity.The pleural effusion was detected by Micro Echo technology.The volumes of effusion were quantified and the tumor cells were counted.Results After implanted of tumor cell, malignant pleural effusion can be detected by Micro Echo technology and observed after autopsy.Chemotherapy drugs such as Cyclophosphamide and Cisplatin can decrease the effusion volumes.Conclusion Pleural effusion model of Lewis lung cancer based on Micro Echo technology can be used to evaluate the efficacy of antitumor drugs.

Development of the high-throughput screening fluorescence polarization assay for HSP90 inhibitor / 中国生化药物杂志

Objective To establish the high-throughput screening fluorescence polarization assay for HSP90 inhibitor.Methods E.coli strain BL21 ( DE3) competent cells were transformed with pET24α( +)-HSP90αplasmid.The cell lysate supernatant was induced to product the soluble protein and purified with Ni-NTA agarose.Western blot analysis was used to identify whether the purified protein is HSP90α.The fluorescence polarization assay for screening HSP90 inhibitors was established and optimized using varying concentrations of recombinant HSP90 protein and molecular probe VER00051001.Meanwhile, the binding activity of GA and NVP-AUY922 for HSP90αwas measured by fluorescence polarization assay.Results HSP90αwas induced expression and purified successfully.The fluorescence polarization assay was performed using 80 nM probe VER00051001 and 2.01μg/mL HSP90α, with the Z factor of 0.83.GA and NVP-AUY922 competed with the probes VER00051001 for binding sites of HSP90, with IC50 of 55 nM and 13 nM, respectively.Conclusion A reliable model was established using fluorescence polarization assay for screening HSP90 inhibitors.

Effect of antiviral therapy on long-term prognosis of patients with decompensated hepatitis B cirrhosis / 临床肝胆病杂志

ObjectiveTo investigate the effect of antiviral therapy on the long-term prognosis of patients with decompensated hepatitis B cirrhosis. MethodsA total of 132 patients with decompensated hepatitis B cirrhosis who were hospitalized in The People′s Hospital of Lezhi County 3 times or more from May 2009 to May 2012 were enrolled in the study. According to the therapies, the patients were divided into control group (51 patients) and observation group (81 patients). All the patients were followed up for 5 years via outpatient service and by telephone. Child-Pugh score and Model for End-Stage Liver Disease (MELD) score were obtained from all patients, and the reduction in HBV DNA, HBV DNA clearance, and liver function parameters were observed. Clinical outcomes were compared at the end of follow-up between the two groups. The t-test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups. ResultsThe observation group had significantly lower incidence rate of liver cancer, rate of liver transplantation, and mortality rate than the control group (χ2=4.32, 4.33, and 4.71, all P＜0.001). After treatment, the observation group showed a significant reduction in HBV DNA level (t=20.60, P＜0.001) and had a significantly lower HBV DNA level than the control group (t=16.40, P＜0.05). After treatment, the observation group had a significantly higher cumulative clearance rate of HBV DNA than the control group (88.89% vs 6.67%, P＜0.05). After treatment, the observation group had a significantly higher serum albumin level and significantly lower levels of total bilirubin (TBil) and alanine aminotransferase (ALT) than the control group (t=6.77, 16.60, and 11.67, all P＜0.001), and both groups showed significant reductions in TBil and ALT after treatment (observation group: t=25.18 and 23.33, both P＜0.001; control group: t=6.15 and 7.20, both P＜0.001). The observation group showed a significant increase in serum albumin level after treatment (t=10.08, P＜0.001). After treatment, the observation group had significantly lower Child-Pugh score and MELD score than the control group (t=2.27 and 2.24, both P＜0.05), both groups showed significant reductions in Child-Pugh score and MELD score (observation group: t=9.18 and 8.17, P＜0.001 and P=0.03; control group: t=2.93 and 3.12, P=0.01 and 0.04). ConclusionLong-term antiviral therapy can improve the long-term prognosis of patients with decompensated hepatitis B cirrhosis and increase the 5-year survival rate.

The anti-tumor activity and molecular mechanisms of an Aurora kinase inhibitor ZLJ213 in suppressing colon cancer growth / 药学学报

The aim of this study is to evaluate anti-tumor activities and mechanism of a novel kinase inhibitor ZLJ213 which targeted Aurora A and vascular endothelial growth factor receptor (VEGFR) in vitro and in vivo against human colon cancer. Results showed that ZLJ213 inhibited cell proliferation and induced cell cycle arrest and apoptosis of HCT1 16 and SW48 cell lines. In HCT116-derived xenograft, ZLJ213 dosed at 100 mg · kg(-1) inhibited tumor growth by 73.24%. The IC50 of ZLJ213 on the expression of p-Aurora A was 0.258 µmol · L(-1) analyzed by ELISA. Under the concentration of 0.08 µmol · L(-1), ZLJ213 could inhibit the activities of Aurora A, Histone H3 and VEGFR of HCT116 and SW48 cell lines. Simultaneously, ZLJ213 induced activation of Caspase 3 and PARP cleavage. Above data suggested that ZLJ213 had the ability to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo in colon cancer, and down-regulate the expression of p-Aurora A and p-VEGFR. ZLJ213 might be a potential therapeutic agent against colon cancer.

Chemical constituents from Morus notabilis and their cytotoxic effect / 药学学报

Une new flavonoids named as notabilisin K (1), together with four known compounds, morusin (2), mulberrofuran A (3), neocyclomorusin (4) and mornigrol F (5) are separated from 95% ethanol extracts of the twigs of Morus notabilis. Compounds 2-5 are separated from this plant for the first time. Notabilisin I, notabilisin J exhibits certain effect against cells of HCT-116, HepG2 and A2780 with IC50 values ranging from 1.47 μmol x L(-1) to 5.46 μmol x L(-1). Morusin exhibits strong effect against five kinds of human cancer cells (BGC823, A2780, HCT-116, HepG2 and NCI-H1650) with IC50 values ranging from 0.74 μmol x L(-1) to 1.58 μmol x L(-1).

Synthesis of novel curcumin mimics and preliminary evaluation for their antitumor activity / 药学学报

Curcumin has been reported to possess antitumor activity with low toxicity. However, the clinical application of curcumin has been significantly limited by its instability and poor metabolic property. In order to overcome these limitations and discover novel small molecules with potential antitumor activity, 29 curcumin mimics were synthesized, which were confirmed by 1H NMR and HR-MS, and their cytotoxic property was evaluated against five human cancer cell lines in vitro. Compounds 16, 18 and 19 exhibited good cytotoxic property, their IC50 value were even below 5 micromol x L(-1) to some cancer cell lines, 5-9 times better than curcumin.

Anti-tumor activity and mechanism of T03 in vitro and in vivo / 药学学报

The purpose of this study is to investigate the activity and mechanism of a new anti-tumor agent T03. MTT and colony formation assay were performed to determine anti-proliferation activity of T03 in vitro. Antitumor activity was observed by Renca xenograft model in vivo. The effect of T03 on cell cycle and apoptosis were measured by FCM analysis. Western blotting was performed to investigate the expression level of proteins in HepG2 cell lines treated with T03. T03 had anti-tumor activity by inhibiting tumor cell growth and colony formation in vitro, especially on hepatocellular carcinoma cells (HCC). At the concentration of 10 micromol x L(-1), T03 induced cell apoptosis and cell cycle arrest in HCC. Moreover, it proved that T03 reduced the tumor weight with the rate of 42.30% without any obviously side effect in Renca xenograft model. At the concentration of 2.0 micromol x L(-1), T03 was able to reduce the level of p-c-Raf (Ser259), and thus blocked Raf/MEK/ERK and AKT signaling in HepG2 cell lines. The result suggested that T03 has the potential to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo, particularly active against HCC, indicating T03 and its analogues may serve as a new anti-cancer drug against hepatocellular carcinoma.

Development of a novel screening assay for inhibitors targeting HIF-1alpha and P300 interaction / 药学学报

Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.

Synthesis and in vitro cytotoxic activities of sorafenib derivatives / 药学学报

A series of novel sorafenib analogues were designed and synthesized. The cytotoxic activities of these compounds were tested in four tumor cell lines. Some of the compounds showed potent antiproliferative activity against the tested cell lines with IC50 = 4-20 micromol x L(-1). Some compounds demonstrated competitive antiproliferative activities to sorafenib against tested cancer cell lines. Among them, compound 7c demonstrated significant inhibitory activities on ACHN, HCT116 and MDA-MB-231 cell lines with IC50 values of 9.01, 4.97, 6.61 micromol x L(-1), respectively.