ABSTRACT
Several studies revealed a similar down-regulation of telomeric repeat binding factor 1 (TRF1) in tumors. We have previously reported the TRFl expression levels were down-regulation in non-small cell lung cancer (NSCLC). The regulation of TRFl localization is proposed to be important for the function and expression. The nuclear localization signal (NLS) and nuclear export signal (NES) are often important clues to localization of protein. The objective of the present study was to investigate the NLS and NES of TRFl in NSCLC patients. Thirty (30) patients with NSCLCs had undergone radical operations in The First Affiliated Hospital, College of Medicine, Zhejiang University. DNA sequences of NLSs and NESs were amplified by PCR. The PCR products were analyzed by DNA sequencing. There were four NLSs of the TRFl protein, including two monopartite and two bipartite NLSs. The NLSs sequences were included in 337KKERRVGTPQSTKKKKESRR356. The exon 8 and exon 9 of TRFl DNA were covered the NLS sequences. The sequences of predicted NESs were 11WMLDFLCLSL86 and 174NLLKLQALAV183, respectively. The exon 1, exon 3 and exon 4 of TRFl were covered the NES sequences. In NSCLCs, there was no a mutation, deletion, or substitution in NLS and NES of TRFl. We conclude that the NLS and NES sequences in NSCLCs patients did not have mutations. Down-expression of TRFl does not indicate gene mutation of NLS and NES in NSCLCs.
Subject(s)
Female , Humans , Male , Carcinoma, Non-Small-Cell Lung/genetics , Down-Regulation/genetics , Lung Neoplasms/genetics , Telomere-Binding Proteins/genetics , Telomeric Repeat Binding Protein 1/genetics , Exons , Gene Expression Regulation, Neoplastic , Nuclear Export Signals/genetics , Nuclear Localization Signals/genetics , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
The present study is to assess the prophylactic effect of quinacrine (QA) , an anti-malarial drug, against heatstroke in rats. Conscious rats were orally given equal volume normal saline or QA (dissolved in normal saline and final dosage for rats was 4.5, 9.0 and 18 mg x kg(-1)). An hour later rats were put into a warm water circulated hot chamber (41.0 +/- 0.5) degrees C. Rectal temperature (core temperature, T(co)) of rats in hot chamber was continuously monitored by a thermocouple. T(co) and survival time of rats showed that QA pre-treatment postponed the hyperthermia, and increased the survival time of rats in hot chamber. Primary striatum neurons' culture from new born rats was maintained with D-MEM and 10% FBS. After immuno-cytochemistry identification with antibodies against neural specific proteins, culture received 20 micromol x L(-1) QA only for 1 h and followed by 43.0 degrees C heat treatment for another hour, or 20 micromol x L(-1) QA for 1 h followed by 43.0 degrees C heat treatment for another hour. Control culture received heat treatment only. Cultures were labeled with the fluorescent indicator DPH and the relative membrane fluidity of neurons was measured with the help of fluorescent polarized spectrophotometer. [3H] Arachidonic acid (AA) labeled membrane of E. Coli cells was used as substrate to determine cPLA2 activity of neurons. Gas chromatography and mass spectrum were also employed to detect on the level of fatty acids level in rat striatum neurons. Results from cells indicated that inhibition of cPLA2, reduction the release of active fatty acids such as AA, and possibly, stabilization of the cell membrane which was disturbed by hot treatment, may contribute to the mechanism underlying heat protection and heatstroke preventive effects of quinacrine.
Subject(s)
Animals , Male , Rats , Cells, Cultured , Corpus Striatum , Pathology , Fatty Acids , Metabolism , Heat Stroke , Metabolism , Hot Temperature , Membrane Fluidity , Neurons , Metabolism , Physiology , Phospholipases A2 , Metabolism , Quinacrine , Pharmacology , Rats, WistarABSTRACT
<p><b>AIM</b>To investigate the effect of hyperthermia on apoptosis of cultivated striatum neurons in the rat.</p><p><b>METHODS</b>After 30 min hyperthermia in 43 degrees, the Ca2+ concentration, the mitochondria membrane potential of the neuron were detected by Laser Scanning Confocal Microscopy (LSCM). The apoptosis of striatum neurons was detected by TUNEL staining.</p><p><b>RESULTS</b>Heat stress at 43 degrees C for 40 min caused an increase in the Ca2+ concentration of striatum neurons and a decrease in the mitochondria membrane potential of the striatum neurons.</p><p><b>CONCLUSION</b>The striatum shows more apoptosis neurons after heat stress.</p>