Tetrandrine, an activator of autophagy, induces autophagic cell death via PKC-α inhibition and mTOR-dependent mechanisms.

Emerging evidence suggests the therapeutic role of autophagic modulators in cancer therapy. This study aims to identify novel traditional Chinese medicinal herbs as potential anti-tumor agents through autophagic induction, which finally lead to autophagy mediated-cell death in apoptosis-resistant cancer cells. Using bioactivity-guided purification, we identified tetrandrine (Tet) from herbal plant, Radix stephaniae tetrandrae, as an inducer of autophagy. Across a number of cancer cell lines, we found that breast cancer cells treated with tetrandrine show an increase autophagic flux and formation of autophagosomes. In addition, tetrandrine induces cell death in a panel of apoptosis-resistant cell lines that are deficient for caspase 3, caspase 7, caspase 3 and 7, or Bax-Bak respectively. We also showed that tetrandrine-induced cell death is independent of necrotic cell death. Mechanistically, tetrandrine induces autophagy that depends on mTOR inactivation. Furthermore, tetrandrine induces autophagy in a calcium/calmodulin-dependent protein kinase kinase-β (CaMKK-β), 5′ AMP-activated protein kinase (AMPK) independent manner. Finally, by kinase profiling against 300 WT kinases and computational molecular docking analysis, we showed that tetrandrine is a novel PKC-α inhibitor, which lead to autophagic induction through PKC-α inactivation. This study provides detailed insights into the novel cytotoxic mechanism of an anti-tumor compound originated from the herbal plant, which may be useful in promoting autophagy mediated- cell death in cancer cell that is resistant to apoptosis.published_or_final_versio

Detection of Polarization in the Cosmic Microwave Background using DASI

We report the detection of polarized anisotropy in the Cosmic Microwave Background radiation with the Degree Angular Scale Interferometer (DASI), located at the Amundsen-Scott South Pole research station. Observations in all four Stokes parameters were obtained within two 3.4 FWHM fields separated by one hour in Right Ascension. The fields were selected from the subset of fields observed with DASI in 2000 in which no point sources were detected and are located in regions of low Galactic synchrotron and dust emission. The temperature angular power spectrum is consistent with previous measurements and its measured frequency spectral index is -0.01 (-0.16 -- 0.14 at 68% confidence), where 0 corresponds to a 2.73 K Planck spectrum. The power spectrum of the detected polarization is consistent with theoretical predictions based on the interpretation of CMB anisotropy as arising from primordial scalar adiabatic fluctuations. Specifically, E-mode polarization is detected at high confidence (4.9 sigma). Assuming a shape for the power spectrum consistent with previous temperature measurements, the level found for the E-mode polarization is 0.80 (0.56 -- 1.10), where the predicted level given previous temperature data is 0.9 -- 1.1. At 95% confidence, an upper limit of 0.59 is set to the level of B-mode polarization with the same shape and normalization as the E-mode spectrum. The TE correlation of the temperature and E-mode polarization is detected at 95% confidence, and also found to be consistent with predictions. These results provide strong validation of the underlying theoretical framework for the origin of CMB anisotropy and lend confidence to the values of the cosmological parameters that have been derived from CMB measurements.Comment: 20 pages, 6 figure

Aptamer-mediated inhibition of Mycobacterium tuberculosis polyphosphate kinase 2

Poster Presentations: P4

Aptamers against polyphosphate kinase 2 (PPK2) from mycobacterium tuberculosis as a potential approach of novel anti-tuberculosis therapy

Structure-Based Screening and Design of Ligands for Protein Targets, CUHK, Hong Kong, 14-19 December 2009

p27 Kip1 promotes migration of metastatic hepatocellular carcinoma cells

Background/Aim: p27 Kip1 (p27) is a member of the Cip/Kip family of cyclin/cyclin-dependent kinase inhibitors (CKIs), and its CKI-independent function regarding cell motility modulation has been discovered. However, it is controversial whether p27 promotes or inhibits cell migration. This study investigates the migration regulatory role of p27 in metastatic hepatocellular carcinoma (HCC) cells. Methods: RNA interference, RhoA-GTP pull-down assay, Western blots, immunostaining, transwell and wound-healing assays were used. Results: High levels of p27 and phosphorylated p27 (Ser10) were detected in metastatic HCC cells, MHCC97L and MHCC97H, when compared with nonmetastatic HCC cells, PLC and Hep3B. We hypothesized that p27 is responsible for metastasis-related migration in HCC cells and tested the hypothesis by using the p27 RNA interference approach. Increased RhoA activity was observed when p27 was knocked down in MHCC97L cells, which further led to stress fiber formation and decreased cell migration and wound healing. Moreover, high p27 and low stathmin expression of metastatic HCC cells indicated that migration inhibition by p27-stathmin interaction might not be the major regulatory pathway in metastatic HCC cells. Conclusion: p27 promotes cell migration in metastatic HCC cells through the regulation of RhoA activity. Copyright © 2008 S. Karger AG.link_to_subscribed_fulltex

Aptamer-mediated inhibition of mycobacterium tuberculosis polyphosphate kinase 2

Inorganic polyphosphate (polyP) plays a number of critical roles in bacterial persistence, stress, and virulence. PolyP intracellular metabolism is regulated by the polyphosphate kinase (PPK) protein families, and inhibition of PPK activity is a potential approach to disrupting polyP-dependent processes in pathogenic organisms. Here, we biochemically characterized Mycobacterium tuberculosis (MTB) PPK2 and developed DNA-based aptamers that inhibit the enzymes catalytic activities. MTB PPK2 catalyzed polyP-dependent phosphorylation of ADP to ATP at a rate 838 times higher than the rate of polyP synthesis. Gel filtration chromatography suggested MTB PPK2 to be an octamer. DNA aptamers were isolated against MTB PPK2. Circular dichroism revealed that aptamers grouped into two distinct classes of secondary structure; G-quadruplex and non-G-quadruplex. A selected G-quadruplex aptamer was highly selective for binding to MTB PPK2 with a dissociation constant of 870 nM as determined by isothermal titration calorimetry. The binding between MTB PPK2 and the aptamer was exothermic yet primarily driven by entropy. This G-quadruplex aptamer inhibited MTB PPK2 with an IC 50 of 40 nM and exhibited noncompetitive inhibition kinetics. Mutational mechanistic analysis revealed an aptamer G-quadruplex motif is critical for enzyme inhibition. The aptamer was also tested against Vibrio cholerae PPK2, where it showed an IC 50 of 105 nM and insignificant inhibition against more distantly related Laribacter hongkongensis PPK2. © 2011 American Chemical Society.link_to_subscribed_fulltex

Notch1-Snail1-E-cadherin pathway in metastatic hepatocellular carcinoma

Notch signaling, a critical pathway for tissue development, also contributes to tumorigenesis in many cancers, but its pathological function in liver cancer is not well defined. In our study, Notch1 expression and its clinicopathological parameters were evaluated in 82 human hepatocellular carcinoma (HCC) patients. Plasmid-based siNotch1 shRNA was transiently or stably transfected into metastatic HCC cells and subsequently evaluated for the effects on orthotopic liver tumor metastasis in a mouse model as well as the effects on downstream pathways. Aberrant high expression of Notch1 was significantly associated with metastatic disease parameters in HCC patients, such as tumor-node-metastasis Stages III-IV and tumor venous invasion. Knocking-down Notch1 reduced cell motility in vitro and orthotopic tumor metastasis from the liver to the lung in vivo in a mouse model. In metastatic HCC cells, abnormal expression of Notch1 was associated with increased expression of Snail1 and repressed expression of E-cadherin; the Notch1-Snail1-E-cadherin association can also be found in HCC patient tumors. Inhibition of Notch1 by shRNA abolished Snail1 expression, which further resulted in the re-establishment of repressed E-cadherin in metastatic HCC cells. Thus, abnormal Notch1 expression was strongly associated with HCC metastatic disease, which might be mediated through the Notch1-Snail1-E-cadherin pathway. Knock-down of Notch1 reversed HCC tumor metastasis in a mouse model. Therefore, these data suggest that effective targeting of Notch signaling might also inhibit tumor metastasis. Copyright © 2011 UICC.link_to_subscribed_fulltex