Effect of Carvacrol on the Biological Behavior of Leukemia Cells and Its Mechanism / 中国实验血液学杂志

OBJECTIVE@#To explore the effect of carvacrol on the biological behavior of leukemia cells and its regulation to circ-0008717/miR-217 molecular axis.@*METHODS@#Human acute lymphoblastic leukemia cells Molt-4 were cultured in vitro, and different concentrations of carvacrol were added to the cells. si-NC and si-circ-0008717 were transfected into Molt-4 cells (si-NC group, si-circ-0008717 group). pcDNA, pcDNA-circ-0008717, anti-miR-NC, anti-miR-217 were transfected into Molt-4 cells and then added to carvacrol-treated cells (carvacrol+pcDNA group, carvacrol+pcDNA-circ-0008717 group, carvacrol+anti-miR-NC group, carvacrol+anti-miR-217 group). MTT, plate clone formation experiment, and flow cytometry were used to detect the viability of the cell, colony formation number, and apoptosis rate of cells, respectively. The RT-qPCR method was used to detect the expression levels of circ-0008717 and miR-217. The dual luciferase reporter gene experiment was used to detect the targeting relationship between circ-0008717 and miR-217.@*RESULTS@#After carvacrol treatment, the cell viability decreased significantly (r=-0.9405), expression level of circ-0008717 decreased (r=-0.9117), colonies formed number decreased (r=-0.9256), while the cell apoptosis rate increased (r= 0.8464), and the expression level of miR-217 increased (r=0.9468). Compared with the si-NC group, the expression level of miR-217 in si-circ-0008717 group increased (P＜0.001), the cell apoptosis rate increased (P＜0.001), while cell viability decreased (P＜0001), the number of colonies formed decreased (P＜0.001). Compared with the carvacrol+pcDNA group, the cell viability of the carvacrol+pcDNA-circ-0008717 group increased (P＜0.001), the number of colonies formed increased (P＜0.001), while the cell apoptosis rate decreased (P＜0.001). circ-0008717 could target miR-217. The cell viability of the carvacrol+anti-miR-217 group increased (P＜0.001), and the number of colonies formed increased (P＜0.001), while the cell apoptosis rate decreased (P＜0001) as compared with the carvacrol+anti-miR-NC group.@*CONCLUSION@#Carvacrol can promote the expression of miR-217 by down-regulating the expression of circ-0008717, thereby reducing the proliferation and cloning ability of leukemia cells and promoting cell apoptosis.

Application of Real-time Quantitative PCR in Detecting Atypical BCR/ABL mRNA Transcripts in Chronic Myelocytic Leukemia / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To detect atypical BCR/ABL mRNA transcript by real-time quantitative PCR in CML patients without e13a2/e14a2,e19a2 or e1a2 transcripts, and investigate its value of clinical application.</p><p><b>METHODS</b>Twelve cases of CML with positive for t(9;22) translocation, but negative for common major and minor breakpoint cluster regions comfirmed by chromosome karyotyping or FISH analysis, were collected from July 2012 to December 2015. These 12 cases were then detected for b2a3(e13a3), b3a3(e14a3), e6a2, e8a2 and e1a3 fusion variants by real-time quantitative PCR.</p><p><b>RESULTS</b>Among 12 cases 4 variant transcripts were detected, including e1a3 in 1 case (8.33%), e8a2 in 2 cases (16.67%), b2a3 in 5 cases (41.67%) and b3a3 in 4 cases (33.33%), with total positivity of 100%, moreover b2a3 and b3a3 were predominant.</p><p><b>CONCLUSION</b>The detecting atypical BCR/ABL mRNA transcripts by real-time quantitative PCR is suitable for the diagnosis of CML negative for P210, P190 and P230 by standard real-time PCR test, and this detection is still the standard and economic method for monitoring minimal residual disease in CML patients with variants of BCR/ABL fusion gene.</p>

Regulatory effect of curcumin on p300 and HDAC1 in B-NHL cells / 中国实验血液学杂志

The purpose of this study was to investigate the effect of curcumin on proliferation of B-NHL Raji cell line and explore the relationship between this effect and regulatory expression of p300 and HDAC1 transcription. The in vitro cultured Raji cells were treated with curcumin at various concentrations (6.25-50 micromol/L) and at different time points (0, 6, 12, 24 and 48 hours), the inhibitory ratio of cell growth was measured by MTT assay, the cell apoptosis rate was detected by flow cytometry with Annexin V-FITC/PI double staining, the changes of p300 and HDAC1 mRNA expression and protein level in Raji cells were determined by RT-PCR and Western blot. The results showed that the curcumin could inhibit Raji cell proliferation in significant time-and concentration-dependent manners, IC50 at 24 hours was 25 micromol/L; the curcumin could induce apoptosis of Raji cells in concentration-dependent manner, apoptosis rate was 14.38%-61.18%. The curcumin significantly inhibited activity and expression of p300 and HDAC1. At IC50 concentration, expression of p300 and HDAC1 mRNA and protein level decreased with time-dependent manner, difference between tested and control groups was significant (P < 0.05). It is concluded that the curcumin can inhibit proliferation of B-NHL Raji cells and promote apoptosis of those cells. Curcumin can inhibit the activity and expression of the transcriptional co-activator p300 and HDAC1, which may be involved in its pharmacological mechanisms on B lymphoma cells.