Circ_0005273 induces the aggravation of pancreatic cancer by targeting KLF12.

OBJECTIVE: The purpose of this study was to explore the potential influences of circ_0005273 and its downstream target KLF12 on the progression of pancreatic cancer. PATIENTS AND METHODS: Relative levels of circ_0005273 and KLF12 in paired pancreatic cancer tissues and normal tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Then, the differences in clinical indicators and prognosis (overall survival and progression-free survival) between pancreatic cancer patients expressing high and low levels of circ_0005273 were compared. After knockdown of circ_0005273 in AsPC-1 and CFPAC-1 cells, viability and migratory ability were assessed by cell counting kit-8 (CCK-8), transwell and wound healing assays. The regulatory effect of circ_0005273 on KLF12 was determined through Western blotting assay. Finally, the interaction between circ_0005273 and KLF12 was tested by dual-luciferase reporter assay. RESULTS: It was found that circ_0005273 was upregulated in pancreatic cancer tissues than that in normal tissues. Besides, pancreatic cancer patients expressing a high level of circ_0005273 had higher incidence rates of lymphatic metastasis and distant metastasis, as well as poor prognosis. Knockdown of circ_0005273 weakened the proliferative and migratory abilities of AsPC-1 and CFPAC-1 cells. KLF12 was the target gene binding to circ_0005273, showing a negative expression correlation between each other. Moreover, the protein level of KLF12 was downregulated by knockdown of circ_0005273. KLF12 was able to abolish the regulatory effects of circ_0005273 on the phenotypes of pancreatic cancer cells. CONCLUSIONS: Circ_0005273 drives proliferative and migratory abilities of pancreatic cancer cells via activating the KLF12, and it is able to predict lymphatic metastasis, distant metastasis and prognosis in pancreatic cancer patients.

MiR-4282 contributes to inhibit pancreatic cancer metastasis by negatively interacting with ABCB5.

OBJECTIVE: To explore the regulatory mechanism of microRNA-4282 (miR-4282) on influencing pancreatic cancer progression by targeting ABCB5. PATIENTS AND METHODS: MiR-4282 and ABCB5 levels in 58 cases of pancreatic cancer and paracancerous tissues were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The influences of miR-4282 on pathological indicators and prognosis in pancreatic cancer patients were analyzed. MiR-4282 overexpression model was established in PANC-1 and BxPC-3 cells by transfection of miR-4282 mimic. Transwell and wound healing assay were conducted to illustrate the role of miR-4282 in influencing cell functions of pancreatic cancer. Bioinformatics analysis and Dual-Luciferase reporter assay were carried out to ascertain the interaction between miR-4282 and ABCB5. RESULTS: MiR-4282 was downregulated in pancreatic cancer samples. Low level of miR-4282 predicted high incidences of lymphatic metastasis and distant metastasis, as well as poor prognosis in pancreatic cancer patients. Overexpression of miR-4282 remarkably inhibited migratory ability in PANC-1 and BxPC-3 cells. MiR-4282 was targeted by ABCB5 through specific binding sites. In pancreatic cancer tissues, ABCB5 level was negatively correlated to that of miR-4282. Overexpression of ABCB5 could abolish the inhibitory effects of overexpressed miR-4282 on the malignant progression of pancreatic cancer. CONCLUSIONS: MiR-4282 is able to inhibit the migratory ability in pancreatic cancer cells by negatively targeting ABCB5, which may become a promising pancreatic cancer biomarker.

Effects of miR-132 on proliferation and apoptosis of pancreatic cancer cells via Hedgehog signaling pathway.

OBJECTIVE: Micro ribonucleic acids (miRNAs) and Hedgehog (Hh) signaling pathway play key roles in the proliferation, migration and invasion of tumor cells. The aim of this study was to investigate the role of miR-132 and Hh signaling pathway in the proliferation and apoptosis of pancreatic cancer cells, and to investigate the possible underlying mechanism. MATERIALS AND METHODS: The expressions of miR-132 and Shh (a ligand of Hh) in clinical pancreatic cancer specimens and pancreatic cancer cell lines were determined by qRT-PCR. Meanwhile, the correlation between the two molecules was analyzed. Pancreatic cancer cell line (MiaPaCe-2a) was transfected with miR-132 mimics and inhibitor. The effects of miR-132 up- and down-regulation on the expressions of miR-132, Shh, Cyclin-D1, cleaved Caspase-3 and cleaved Caspase-9 were detected. In addition, the exact role of miR-132 in the proliferation, apoptosis and distribution of MiaPaCe-2a cells were investigated. RESULTS: The expression level of miR-132 in pancreatic cancer specimens and pancreatic cancer cell lines was significantly elevated when compared with that of control group. Meanwhile, miR-132 expression was negatively correlated with the expression level of Shh. Moreover, transfection with miR-132 mimics evidently up-regulated miR-132 expression. Moreover, miR-132 up-regulation significantly decreased the mRNA and protein expressions of Shh, facilitated the proliferation of MiaPaCe-2a cells, reduced the protein expressions of Cyclin-D1, cleaved Caspase-3 and cleaved Caspase-9, and suppressed cell apoptosis. On the contrary, miR-132 inhibitor transfection significantly inhibited the proliferative activity of MiaPaCe-2a cells, decreased the proportion of cells in G1 phase, and increased the proportion of cells in G2/M phase. CONCLUSIONS: MiR-132 promotes proliferation and inhibits apoptosis of pancreatic cancer cells through Hh signaling pathway.

Long non-coding RNA CCAT1 promotes the migration and invasion of prostate cancer PC-3 cells.

OBJECTIVE: To investigate the effect of long non-coding RNA CCAT1 on the proliferation, migration, and invasion of prostate cancer PC-3 cells. PATIENTS AND METHODS: The expression of CCAT1 was detected by Real-time PCR. The effect of CCAT1 down-regulation on the proliferation of PC-3 cells was observed by MTT assay. The regulatory of CCAT1 low-expression on the migration ability of PC-3 cells was investigated by transwell assay. The influence of decreased CCAT1 on the invasion ability of PC-3 cells was detected by  Matrigel invasion assay. RESULTS: Increased CCAT1 was significantly related to lymph node metastasis in prostate cancer. Low-expression of CCAT1 could suppress cell proliferation. Knockdown of CCAT1 inhibited the migration of PC-3 cells. Down-regulation of CCAT1 attenuated the invasion of PC-3 cells. CONCLUSIONS: CCAT1 promoted the growth and the metastasis of prostate cancer. Our findings might provide a potential target for the diagnosis and treatment of prostate cancer.

Lattice-distortion Induced Magnetic Transition from Low-temperature Antiferromagnetism to High-temperature Ferrimagnetism in Double Perovskites A2FeOsO6 (A = Ca, Sr).

High-temperature insulating ferrimagnetism is investigated in order to further reveal its physical mechanisms, as well as identify potentially important scientific and practical applications relative to spintronics. For example, double perovskites such as Sr2FeOsO6 and Ca2FeOsO6 are shown to have puzzling magnetic properties. The former is a low-temperature antiferromagnet while the latter is a high-temperature insulating ferrimagnet. In order to understand the underlying mechanisms, we have investigated the frustrated magnetism of A2FeOsO6 by employing density functional theory and maximally-localized Wannier functions. We find lattice distortion enhances the antiferromagnetic nearest-neighboring Fe-O-Os interaction, however weakens the antiferromagnetic interactions via the Os-O-O-Os and Fe-O-Os-O-Fe paths, so is therefore responsible for the magnetic transition from the low-temperature antiferromagnetism to the high-temperature ferrimagnetism as the decrease of the A(2+) ion radii. Also discussed is the 5d(3)-3d(5) superexchange. We propose that such superexchange is intrinsically antiferromagnetic instead of ferromagnetic as previously thought. Our work clearly illustrates the magnetic frustration can be effectively relieved by lattice distortion, thus paving the way for tuning of complex magnetism in yet other 3d-5d (4d) double perovskites.

Rotenone impairs autophagic flux and lysosomal functions in Parkinson's disease.

BACKGROUND: Rotenone is an environmental neurotoxin that induces accumulation of α-synuclein and degeneration of dopaminergic neurons in substantia nigra pars compacta (SNpc), but the molecular mechanisms are not fully understood. We investigated whether rotenone induced impairment of autophagic flux and lysosomal functions. METHODS: Autophagy flux, accumulation of α-synuclein, lysosomal membrane integrity and neurodegeneration were assessed in the rotenone-treated rat model and PC12 cells, and the effects of the autophagy inducer trehalose on rotenone's cytotoxicity were also studied. RESULTS: Rotenone administration significantly reduced motor activity and caused a loss of tyrosine hydroxylase in SNpc of Lewis rats. The degeneration of nigral dopaminergic neurons was accompanied by the deposition of α-synuclein aggregates, autophagosomes and redistribution of cathepsin D from lysosomes to the cytosol. In cultured PC12 cells, rotenone also induced increases in protein levels of α-synuclein, microtubule-associated protein 1 light chain 3-II, Beclin 1, and p62. Rotenone increased lysosomal membrane permeability as evidenced by leakage of N-acetyl-beta-d-glucosaminidase and cathepsin D, the effects were blocked by reactive oxygen species scavenger tiron. Autophagy inducer trehalose enhanced the nuclear translocation of transcription factor EB, accelerated the clearance of autophagosomes and α-synuclein and attenuated rotenone-induced cell death of PC12 cells. Meanwhile, administration of trehalose to rats in drinking water (2%) decreased rotenone-induced dopaminergic neurons loss in SNpc. CONCLUSIONS: These studies indicate that the lysosomal dysfunction contributes to rotenone's neurotoxicity and restoration of lysosomal function could be a new therapeutic strategy for Parkinson's disease.

(-)-Discretamine, a selective alpha 1D-adrenoceptor antagonist, isolated from Fissistigma glaucescens.

1. The selectivity of (-)-discretamine for alpha 1-adrenoceptor subtypes was investigated by use of functional and binding studies in rat vas deferens, spleen and aorta, and in cultured DDT1MF-2 and A10 cells. 2. In prostatic portions of rat vas deferens, the competitive antagonists (-)-discretamine, 5-methylurapidil (5-MU) and prazosin inhibited contractions to noradrenaline (NA) with pA2 values of 6.21, 8.71 and 9.27, respectively. The irreversible antagonist, chloroethylclonidine (CEC, 100 microM) failed to affect contractions to NA while nifedipine (1 microM) blocked them almost completely. 3. In rat spleen, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to phenylephrine with pA2 values of 6.44, 7.19 and 9.45, respectively. CEC (100 microM) significantly reduced the maximum contraction to phenylephrine while nifedipine (1 microM) did not affect it. 4. In rat aorta, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to NA with pA2 values of 7.60, 8.00 and 9.40, respectively. CEC also antagonized the contractions to NA in a competitive manner with a pA2 value of 6.10. 5. The specific binding of [3H]-prazosin to DDT1MF-2 and A10 cells was concentration-dependent and saturated at 3-5 nM with KD values of 0.24 +/- 0.02 and 0.20 +/- 0.02 nM, respectively. (-)-Discretamine,5-MU, CEC and prazosin inhibited specific [3H]-prazosin binding to DDTIMF-2 and AlO cells in a concentration-dependent manner with ICso values of 390.8 +/- 20.6, 43.6 +/- 3.9, 200.0 +/- 30.0 and 0.8 +/- 0.1 nM, respectively in DDTIMF-2 cells, and 25.0 +/- 3.2, 8.6 +/- 1.4, 1000.0 +/- 30.8 and 0.52 +/- 0.03 nM, respectively in AlO cells.6. Pretreatment of Al0 cells with CEC (10 MicroM) for 30 min and then washed out thoroughly, reduced specific [3H]-prazosin binding by 30%. The CEC-insensitive [3H]-prazosin binding was inhibited by(-)-discretamine with an IC50 value of 7.0 +/- 0.3 nM.7. 5-MU (100 nM), CEC (1 MicroM) and prazosin (10 nM) markedly inhibited NA (3 MicroM)-induced [3H]-inositol monophosphate formation in DDTIMF-2 and A1O cells, while (-)-discretamine (100 nM)inhibited NA-induced [3H]-inositol monophosphate formation only in AlO cells.8. In conclusion, (-)-discretamine is a selective alpha lD-adrenoceptor antagonist in vascular smooth muscle.Its selectivity among various al-adrenoceptor subtypes is alpha 1A: alpha1B: alpha1D =0.04:0.07:1.0.

Mechanisms of vasorelaxation induced by N-allylsecoboldine in rat thoracic aorta.

N-Allylsecoboldine was shown to be the most effective of several boldine derivatives that were tested for their vasorelaxing effect on the rat aorta. In KCl (60 mmol/l) medium, Ca2+ (0.03-3 mmol/l)-induced vasoconstriction was inhibited, concentration-dependently, by N-allylsecoboldine. The IC50 for N-allylsecoboldine was calculated to be about 4 mumol/l (for a Ca2+ concentration of 1 mmol/l). The vasorelaxant effect on KCl-induced responses was more pronounced at 60 mmol/l KCl than at 15 mmol/l KCl. Contraction of rat aorta in response to phenylephrine (0.01-100 mumol/l) was concentration-dependently inhibited by N-allylsecoboldine and by verapamil (3-30 mumol/l), while contraction in response to B-HT 920, serotonin or PGF2 alpha was not affected. This relaxing effect of N-allylsecoboldine persisted in endothelium-denuded aorta. In cultured A10 vascular smooth muscle cells, N-allylsecoboldine and verapamil displaced the binding of [3H]-prazosin (Ki values = 0.4 +/- 0.2 and 0.6 +/- 0.2 mumol/l, respectively). The increase of inositol monophosphate caused by phenylephrine in rat aorta was completely suppressed by chloroethylclonidine, but only slightly inhibited by N-allylsecoboldine and by verapamil. Glibenclamide or charybdotoxin did not affect the relaxation induced by N-allylsecoboldine of aortic rings precontracted with phenylephrine. Neither the cGMP nor the cAMP content was changed by N-allylsecoboldine. We conclude that N-allylsecoboldine relaxes the rat aorta by blocking Ca2+ channels and that it also has an antagonistic effect at alpha 1-adrenoceptors.

[UV-spectrophotometric analysis of the component of antondin injection by the Monte Carlo method of area fitting].

The Monte Carlo method is a kind of unique calculation method. It can resolve not only the question of define evaluation, but also that of random evaluation. The Monte Carlo area fitting method was applied to UV-spectrophotometry of the multicomponent complex preparations without previous separation. The area of each component of complex preparation within a suitable wavelength range of standard spectrogram was multiplied by certain coefficients, and then the products were obtained to fit the area of the measured multicomponent complex preparations. The method was used to simultaneous determination of barbital, amidopyrine and antipyrine in antondin injection. The experimental data were measured and treated by UV-190 spectrophotometer and microcomputer IBM-PC. The average recoveries of barbital, amidopyrine and antipyrine were 99.1 +/- 1.1% (CV), 99.6 +/- 1.3% (CV), and 99.7 +/- 1.0% (CV), respectively.