ABSTRACT
OBJECTIVE@#To investigate the effect of ursane triterpenoids 3β,19α-dihydroxyursu-12-ene-23,28-dicarboxylic acid (Rotundioic acid, RA) on the sensitivity of adriamycin-resistant K562 cells (K562/ADM Cell) anti-tumor drug, and to explore the effect and mechanism of RA on the multidrug resistance of K562/ADM cells.@*METHODS@#CCK-8 method was used to detect the effect of RA on the sensitivity of K562 cells and K562/ADM cells to anti-tumor drug. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression level of mRNA and the protein in K562 and K562/ADM cells, and the effect of RA on the expression of MDR1 mRNA and P-gp in K562/ADM cells was also detected; Western blot was used to detect the expression of p-JNK, p-p38 and p-ERK1/2 in K562/ADM cells.@*RESULTS@#RA could increased the sensitivity of K562/ADM cells to adriamycin(the reversal factor was 1.61 times), the difference showed statistically significantly (P<0.05); the resistance factor of K562/ADM to ADM was 41.76 times. The expression of MDR1 mRNA in K562 cells was extremely low, and the protein product P-glycoprotein (P-gp) was almost not expressed; MDR1 mRNA and P-gp in K562/ADM cells were highly expressed; RA could down-regulate the expression levels of MDR1 and P-gp in K562/ADM cells. In addition, RA could upregulate the phosphorylation levels of p38 and ERK1/2 in K562/ADM cells, but it has no effect on the expression of p-JNK.@*CONCLUSION@#RA may participate in the regulation of MAPK signaling pathway by upregulating the expression levels of p-p38 and p-ERK1/2 in K562/ADM cells, and thus inhibit the transcription and translation levels of MDR1, and finally reverse the multidrug resistance of leukemia cells.
Subject(s)
Humans , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1 , Drug Resistance, Multiple , Drug Resistance, Neoplasm , K562 CellsABSTRACT
CRISPR/Cas genome editing technology is a newly developed powerful tool for genetic manipulation, which can be used to manipulate the genome at specific locations precisely, to restore the function of genetic defect cells, and to develop various disease models. In recentl years, with the advances of precise genome manipulation, CRISPR/Cas technology has been applied to many aspects of diseases research and becomes an unique tool to investigate gene function and discover new therapeutic targets for genetic diseases. Nowadays, CRISPR/Cas technology has been a hot research point in agriculture, graziery, biotechnology and medicine. This review focuses on the recent advances in CRISPR/Cas technology and its application in hematological diseases.
Subject(s)
Humans , CRISPR-Cas Systems , Gene Editing , Genetic Techniques , Genome , Hematologic Diseases , PhenotypeABSTRACT
<p><b>OBJECTIVE</b>To explore the expression and significance of NLR family, pyrin domain containing 3 (NLRP3), apoptosis associated speck like protein containing a CRAD (ASC) and absent in melanoma 2 (AIM2) of patients with acute leukemia.</p><p><b>METHODS</b>The petipheral blood samples of 19 patients with ALL and 41 patients with ANLL as the AL group (each 20 cases of newly diagnosed, relapsed and complete remission group) and 20 cases of non-hematologic malignancies as the control group were collected from July 2013 to July 2014 in the First Affiliated Hospital of Gannan Medical University. The expression levels of NLRP3, ASC and AIM2 in peripheral blood plasma were determined by ELISA.</p><p><b>RESULTS</b>The expression levels of NLRP3, ASC and AIM2 in plasma of control and AL complete remission groups were significantly higher than those in newly diagnosed and relapsed groups, and were with statistical significance (P < 0.05), but there were no statistical signifirance between ALL and ANLL groups (P > 0.05).</p><p><b>CONCLUSION</b>The expression of NLRP3, ASC and AIM2 is down-regulated in the patients with acute leukemia, which maybe play a role of anti-leukemia, and provide a laboratory evidence for diagnosis and treatment of patients with acute leukemia.</p>
Subject(s)
Humans , Acute Disease , CARD Signaling Adaptor Proteins , Carrier Proteins , Blood , Genetics , Case-Control Studies , Cytoskeletal Proteins , Blood , Genetics , DNA-Binding Proteins , Blood , Genetics , Leukemia , Blood , Genetics , Leukemia, Myeloid, Acute , Blood , Genetics , NLR Family, Pyrin Domain-Containing 3 ProteinABSTRACT
Long non-coding RNA (LncRNA) is defined as a class of transcripts more than 200 nucleotides in length and without the protein-coding function. It has been found for years, however, that little is known about the potential role of LncRNA in humans. But recent studies showed that LncRNA can regulate the coding-gene expression and participate in effects of human body. Accumulating evidence demonstrated that LncRNA are involved in cancer incidence, development and progression.With further exploration on the mechanisms of tumors, the relationship between the long non-coding RNA and hematological malignancies increasingly become a hot research. This review focuses on the mechanisms of LncRNA in hematological malignancies.
Subject(s)
Humans , Hematologic Neoplasms , Genetics , RNA, Long Noncoding , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To explore the expression of high mobility group box protein 1 (HMGB1) and nuclear factor-kappa B (NF-κB) in patients with acute leukemia and its significance.</p><p><b>METHOD</b>20 samples of bone marrow and peripheral blood from each acute leukemia groups (newly diagnozed, relapsed and complete remission groups) and 20 samples as control from patients with no-hematologic malignancies were collected. The expression level of HMGB1 in peripheral blood plasma was determined by ELISA; HMGB1 and NF-κB level in mononuclear cells were examined by RT-PCR. Western blot was used to determine HMGB1 and NF-κB protein levels. HMGB1 and NF-κB in bone marrow smears were determined by immnohistochemistry method (IHC).</p><p><b>RESULTS</b>The expression level of HMGB1 obviously increased in patients of newly diagnosed and relapsed groups, as compared with control group there was statistical significance (P < 0.05), but there was no obvious difference in expression level of HMGB1 between complete remission group and control group (P > 0.05). The expression level of HMGB1 and NF-kB in monnuclear cells of bone marrow in newly-diagnosed group and relapsed group was significantly higher than that in control group (P < 0.05), but the expression levels of HMGB1 and NF-kB in complete remisson group did not change (P > 0.05). The results of immnohistochemistry method indicated that the possitive expression of HMGB1 and NF-kB maily was found in bone marrow smears of newly diagnosed and relapsed groups.</p><p><b>CONCLUSION</b>HMGB1 is overexpressed in acute leukemia, which may be involved in the occurrence and development of acute leukemia by activating the NF-κB signaling pathway, HMGB1 may be a important index for observing therapeutic effectiveness and predicting recurrence of acute leukemia.</p>
Subject(s)
Humans , Acute Disease , Blotting, Western , Bone Marrow , Case-Control Studies , HMGB1 Protein , Metabolism , Leukemia , Diagnosis , Metabolism , NF-kappa B p50 Subunit , Metabolism , Remission Induction , Signal TransductionABSTRACT
<p><b>OBJECTIVE</b>To investigate the structure and function of the N-terminal region (NTR) of death receptor 5 (DR5).</p><p><b>METHODS</b>A series of deletions of the DR5 extracellular domain (DR5-ECD) proteins were expressed in E.coli. and purified by affinity chromatography. The binding ability of these deletant proteins to AD5-10, a mouse anti-human DR5 monoclonal antibody, was evaluated by immunoblotting and ELISA.</p><p><b>RESULTS</b>Recombinant DR5-ECD proteins containing the NTR were recognized and bound by AD5-10, while the other deletant proteins without the NTR failed to interact with AD5-10.</p><p><b>CONCLUSION</b>There is an AD5-10 targeting site in the NTR of DR5, which may play a role in developing novel immunotherapies for cancers.</p>