1.Expression of HSP70 in peripheral lymphocytes of the patients with allergic rhinitis.
Lisi, LIU ; Chengfeng, XIAO ; Ming, ZHANG ; Lei, CHENG ; Efen, WANG ; Tangchun, WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(3):310-2
The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49 +/- 15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89 +/- 10.13 IOD, P < 0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR.
Gene Expression Regulation
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HSP70 Heat-Shock Proteins/*biosynthesis
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HSP70 Heat-Shock Proteins/blood
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HSP70 Heat-Shock Proteins/genetics
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Lymphocytes/*metabolism
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Rhinitis, Allergic, Seasonal/*blood
2.Identification of heat shock protein hsp70 family genes from Rana amurensis and its expression profiles upon infection.
Tingting LIU ; Jingjing GUO ; Zhaodong CHEN ; Yufen LIU ; Legang JING ; Peng LIU ; Wenge ZHAO
Chinese Journal of Biotechnology 2023;39(4):1710-1730
Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.
Heat-Shock Proteins/genetics*
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Phylogeny
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Amino Acid Sequence
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HSP70 Heat-Shock Proteins/metabolism*
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Stress, Physiological
3.Effects of occupational microwave irradiation on heat shock protein 70 expressions in rat hippocampus.
Yu-tong HAO ; Li-ping PEI ; Chun-hai CHEN ; Xue-sen YANG ; Guang-bin ZHANG ; Zhao-hui DENG ; Zheng-ping YU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2009;27(9):553-556
OBJECTIVETo study the change of heat shock protein (HSP)70 expression after exposure to occupational microwave in rats hippocampus, and explore the role of HSP70 in the mechanism of bio-effect of microwave irradiation.
METHODSThe animal model was established by whole body exposures in 90, 5 W/cm(2) microwave irradiation field for 20 min in rats. Changes of the mRNA of hsp70 expressions in rat hippocampus at different time were studied by RT-PCR, and the protein change by Western blot.
RESULTSThe mRNA and protein expression of hsp70 in rat hippocampus increased after 90 W/cm(2) and 5 W/cm(2) microwave irradiation for 20 min. The anal temperature and the value of SAR increased significantly. These changes were positively correlated with power and irradiation time of microwave. The results indicated that microwave irradiation led to HSP70 syntheses effectively.
CONCLUSIONMicrowave irradiation can obviously induce the thermal effect and activate HSP70, and initiate the endogenous protective mechanism of central nervous system.
Animals ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Hippocampus ; metabolism ; radiation effects ; Microwaves ; adverse effects ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar
4.Expression of heat stress protein 70 mRNA in patients with chronic obstructive pulmonary disease and its significance.
Jianping, ZHAO ; Jungang, XIE ; Yongjian, XU ; Zhenxiang, ZHANG ; Ning, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2005;25(1):20-3
The effects of cigarette smoke extract (CSE) on the expression of heat stress protein 70 (Hsp70) in human bronchi smooth muscle cells were investigated in vitro, and the changes in Hsp70 mRNA in the patients with chronic obstructive pulmonary disease and their significance were explored. Human bronchi smooth muscle cells were cultured with CSE at the different concentrations. The expression of Hsp70 mRNA and Hsp70 was detected by reverse translation-polymerase chain reaction (RT-PCR) and Western blotting respectively. Levels of Hsp70 mRNA and Hsp70 in lymphocytes from 20 patients with COPD and 20 healthy smoking control subjects were measured by RT-PCR and Western blotting. The results showed the expression of both Hsp70 mRNA and Hsp70 was decreased conformably in human bronchi smooth muscle cells treated with CSE at certain concentration in vitro. The A values of the Hsp70 mRNA expression were 0.24 +/- 0.11 and 0. 42 +/- 0.13 respectively in COPD patients and healthy smoking controls with the difference being significant (P < 0.01). There was also significant difference in the A values of the Hsp70 expression between COPD patients and healthy smoking controls (20.9 +/- 9.9 vs 44.8 +/- 15.3, P < 0.01). The levels of Hsp70 mRNA had strongly positive correlation with Hsp70 protein (r = 0.85, P < 0.01). It was suggested that the expression of Hsp70 mRNA was in concordance with the expression of Hsp70, which could provide a basis on the study of Hsp70 gene regulation and Hsp70 gene in the development of COPD.
Bronchi/metabolism
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Bronchi/pathology
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Cells, Cultured
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HSP70 Heat-Shock Proteins/*biosynthesis
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HSP70 Heat-Shock Proteins/genetics
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Muscle, Smooth/cytology
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Pulmonary Disease, Chronic Obstructive/*metabolism
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RNA, Messenger/biosynthesis
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RNA, Messenger/genetics
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Smoking
5.Construction of eukaryotic expression vector of glucose-regulated protein 75 gene deletion mutant and its expression in PC12 cells..
Wei-Wei GUO ; Ling YANG ; Xiao-Yu LIU ; Wen LIU ; Ji ZUO
Acta Physiologica Sinica 2009;61(6):533-538
Glucose-regulated protein 75 (Grp75) binds to p53 and inhibits its nuclear translocation, and thus plays a role in cell protection. To investigate whether the binding of Grp75 and p53 would influence the viability of cells, we constructed the eukaryotic expression vector of Grp75 deletion mutant. The deletion mutant gene was obtained by SOE-PCR (gene splicing by overlap extension) and then linked to the pcDNA3.0 vector. The constructed specific expression vector, pcDNA3.0/Grp75(Δ253-282), was identified by restriction enzymes and sequencing. Then we used liposome to transfect the specific vector into PC12 cells. The stable cell strain PC12/Grp75(Δ253-282)(+) was selected by G418 (1 mg/mL). Semi-quantitative RT-PCR and Western blot showed that Grp75 mRNA and protein expressions in PC12/Grp75(Δ253-282)(+) cells were higher than those in PC12 cells. The viability of cells undergoing 0 h, 3 h, 9 h, 18 h and 36 h of glucose deprivation respectively was measured by MTT assay. The results showed that the cell viability of PC12/Grp75(+) group was significantly higher than that of the other two groups, and the cell viability of PC12/Grp75(Δ253-282)(+) group was significantly higher than that of the PC12 group (P<0.05). Hoechst33324 staining was employed to detect cell apoptosis and the results were consistent with the MTT assay results. Western blot results indicated that the expression of p53 in PC12/Grp75(+) cells was lower than those in the other two groups, which might be due to the overexpression of Grp75. These results suggest that the protective role of Grp75 is partly associated with its binding to p53. The above results suggest that Grp75 deletion mutation could to some extent reduce the viability of cells.
Animals
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Gene Deletion
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Genetic Vectors
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HSP70 Heat-Shock Proteins
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genetics
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metabolism
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Membrane Proteins
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genetics
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metabolism
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Mutant Proteins
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genetics
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metabolism
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PC12 Cells
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Rats
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Transfection
6.Expression of HSP70 in peripheral lymphocytes of the patients with allergic rhinitis.
Lisi LIU ; Chengfeng XIAO ; Ming ZHANG ; Lei CHENG ; Efen WANG ; Tangchun WU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(3):310-312
The expression levels of heat shock protein 70 (HSP70) from peripheral lymphocytes of the patients with allergic rhinitis (AR) and the clinical implication were investigated. In the morning, 3 ml of fasting venous blood was taken out. The lymphocytes were isolated by using Ficoll-Hypaque and the expression of HSP70 in the lymphocytes was detected by using Western blot. In the AR patients the HSP70 level (41.49 +/- 15.77 integrated optical density, IOD) were significantly higher than that in the control group (23.89 +/- 10.13 IOD, P < 0.05). Western blot demonstrated that HSP70 bands in AR patients were more intensive than those in the control group. It was concluded that the elevated HSP70 level in peripheral lymphocytes of the AR patients might contribute to the development of AR.
Adult
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Female
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Gene Expression Regulation
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HSP70 Heat-Shock Proteins
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biosynthesis
;
blood
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genetics
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Humans
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Lymphocytes
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metabolism
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Male
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Rhinitis, Allergic, Seasonal
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blood
7.Cloning and expression analysis of HSP70 gene from Dendrobium officinale under low temperature stress.
Dong-Bin LI ; Han-Hui GAO ; Jin-Ping SI ; Yu-Qiu ZHU
China Journal of Chinese Materia Medica 2013;38(20):3446-3452
OBJECTIVETo investigate HSP70 gene expression from Dendrobium officinale under low temperature stress, which will provide the molecular biological foundation for breeding the low temperature resistant strain.
METHODHSP70 gene full length cDNA was cloned by rapid amplification of cDNA ends (RACE) on the basis of HSP70 gene fragment sequences, and the structure and function of HSP70 gene were deduced. The expression of HSP70 under low temperature stress was detected by RT-PCR.
RESULTThe full length of HSP70 gene cDNA was 2 296 bp containing a 1 944 bp open reading frame (ORF) that encoded a protein of 647 amino acids. Its amino acids sequence had typical HSP70 characteristics and high homology with other plant's HSP70. Cold stress expression analysis showed that expression of the HSP70 gene could be induced by low temperature.
CONCLUSIONThe HSP70 gene of D. officinale was successfully cloned and reported for the first time which proved that the expression could be induced by low temperature. The cloning of HSP70 gene provides a stable foundation for further study of D. officinale cultivation and the breeding of the cold resistance strains.
Amino Acid Sequence ; Cloning, Molecular ; Cold Temperature ; Dendrobium ; classification ; genetics ; metabolism ; HSP70 Heat-Shock Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment
8.Influence of electroacupuncture on the mRNA of heat shock protein 70 and 90 in brain after cerebral ischemia/reperfusion of rats.
Ning SUN ; Jing SHI ; Liling CHEN ; Xiaochun LIU ; Xinmin GUAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2003;23(2):112-115
To explore the anti-apoptotic role of electroacupuncture (EA) and its molecular mechanisms after cerebral ischemia/reperfusion (IR) of rats, by using animal model of middle cerebral artery occlusion (MCAO), the changes of the cleavage of PARP were observed by Western blot and the mRNA of heat shock protein (Hsp) 70 and Hsp90 beta detected by competitive RT-PCR after cerebral IR and EA treatment. The results were as follows: (1) The cleavage of PARP was increased in ischemic hippocampus, and EA treatment could attenuate the level of the cleavage remarkably; (2) The mRNA expression of Hsp70 was increased in the ischemic cortex and hippocampus and was further increased after EA treatment; (3) The mRNA expression of Hsp90 beta was decreased in ischemic cortex and hippocampus and the decrease was relatively slight after EA treatment. The above results demonstrated EA treatment could protect neurons from apoptosis after cerebral IR. One of the molecular mechanisms was the promotion of the inducible expression of Hsp70 and the improvement of the inhibition of the expression of Hsp90.
Animals
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Apoptosis
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Brain
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metabolism
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Brain Ischemia
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genetics
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metabolism
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Electroacupuncture
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Female
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HSP70 Heat-Shock Proteins
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biosynthesis
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genetics
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HSP90 Heat-Shock Proteins
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biosynthesis
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genetics
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Male
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RNA, Messenger
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biosynthesis
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genetics
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Reperfusion Injury
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genetics
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metabolism
9.What we know about ST13, a co-factor of heat shock protein, or a tumor suppressor?
Zheng-zheng SHI ; Jia-wei ZHANG ; Shu ZHENG
Journal of Zhejiang University. Science. B 2007;8(3):170-176
This article is to summarize the molecular and functional analysis of the gene "suppression of tumorigenicity 13" (ST13). ST13 is in fact the gene encoding Hsp70 interacting protein (Hip), a co-factor (co-chaperone) of the 70-kDa heat shock proteins (Hsc/Hsp70). By collaborating with other positive co-factors such as Hsp40 and the Hsp70-Hsp90 organizing protein (Hop), or competing with negative co-factors such as Bcl2-associated athanogen 1 (Bag1), Hip facilitates may facilitate the chaperone function of Hsc/Hsp70 in protein folding and repair, and in controlling the activity of regulatory proteins such as steroid receptors and regulators of proliferation or apoptosis. Although the nomenclature of ST13 implies a role in the suppression of tumorigenicity (ST), to date available experimental data are not sufficient to support its role in cancer development, except for the possible down-regulation of ST13 in gastric and colorectal cancers. Further investigation of this gene at the physiological level would benefit our understanding of diseases such as endocrinological disorders, cancer, and neurodegeneration commonly associated with protein misfolding.
Adenosine Triphosphate
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metabolism
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Animals
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Carrier Proteins
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chemistry
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genetics
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physiology
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Cloning, Molecular
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HSP70 Heat-Shock Proteins
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metabolism
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Humans
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Protein Folding
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Tumor Suppressor Proteins
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chemistry
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genetics
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physiology
10.Effects of heat shock protein 70-2 gene polymorphisms on the transcription of HSP 70-2 mRNA and the translation of HSP 70 protein in lung cancer.
Hao-quan LU ; Yu-zhen WANG ; Peng-hui SUN ; Shou-pei LIANG ; Jie LI ; Xiao-long WANG ; Dong XU ; Wu YAO ; Yi-ming WU ; Fang ZHOU
Chinese Journal of Preventive Medicine 2012;46(5):452-455
OBJECTIVEThis study aimed to investigate the effects of gene polymorphism of heat shock protein 70-2 (HSP 70-2) 1267A/G on the mRNA level HSP 70-2 mRNA and the protein level HSP 70 in human lung cancer.
METHODSForty six lung cancer patients diagnosed histopathologically between February and August 2008 from a hospital in zhengzhou were enrolled as the subjects in this study. Gene polymorphism of HSP 70-2 1276A/G in 46 patients with lung cancer was detected by PCR-RFLP. The mRNA levels of HSP 70-2 mRNA and the protein levels of HSP 70 in lung tissue and para-cancerous tissues of these subjects were determined by RT-PCR and Western blotting respectively.
RESULTSThe expression levels of HSP 70-2 mRNA (1.02 ± 0.30) and HSP 70 protein (0.44 ± 0.12) in the lung cancer tissues was significantly higher than that in para-cancerous tissues (0.19 ± 0.04, 0.12 ± 0.02). The relative levels of HSP 70-2 mRNA in the subjects with AA genotype (1.32 ± 0.22) were significantly higher than the patients with AG genotype or GG genotype (0.95 ± 0.17, 0.70 ± 0.16) at the site of 1267 (A/G) (P < 0.01); however, the relative protein levels of HSP 70 were 0.47 ± 0.13 (AA genotype), 0.42 ± 0.11 (AG genotype), 0.45 ± 0.11 (GG genotype), respectively, which showed no statistically significant difference (P > 0.05).
CONCLUSIONThe polymorphism of HSP 70-2 1267 (A/G) is highly associated with the transcription level of HSP 70-2 mRNA, but not with the expression level of HSP 70 protein.
Adenocarcinoma ; genetics ; metabolism ; pathology ; Female ; Genotype ; HSP70 Heat-Shock Proteins ; genetics ; Humans ; Lung ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Neoplasm Staging ; Polymorphism, Single Nucleotide ; RNA, Messenger ; genetics