Background: Acute myeloid leukemia (AML) is a common hematopoietic tumor with extremely high morbidity and mortality. This study was designed to explore candidate genes that were related to the poor prognosis of AML patients and analyze their relationship with drug sensitivity. Methods: Microarray databases were performed to screen the differentially expressed genes (DEGs). DAVID 6.8 was used for further functional enrichment analysis. The protein-protein interaction (PPI) network was constructed through STRING website and Cytoscape tool. Then, we analyzed and explored the mRNA transcription level, prognosis correlation, and drug sensitivity of the candidate genes in AML via multiple acknowledged databases including the GEPIA, BloodSpot, EMBL-EBI, UALCAN, LinkedOmics, and GSCALite databases. Results: A total of 181 up-regulated DEGs were screened. Three candidate genes (MAP2K3, LST1, and CYTH4) related to poor outcomes of AML patients were identified. Meanwhile, the high expression levels of the three genes were verified in AML patients and AML cell lines, the expression differences of three genes at AML different subtypes were demonstrated. Drug sensitivity analysis displayed the expression levels of MAP2K3, LST1, and CYTH4 were negatively related to drug resistance, indicating that the three genes were sensitive to certain small-molecule drugs (including targeted drugs and non-targeted drugs). Conclusion: In summary, MAP2K3, LST1, and CYTH4 may be potential prognostic indicators for AML, and may be associated with the sensitivity of certain small molecule drugs.

Background Acute myeloid leukemia (AML) is a common hematopoietic malignancy and have unsatisfactory prognosis. Our study aimed to identify hub genes in AML and explore potential biomarkers through integrated bioinformatics. Methods Microarray datasets were analyzed to screen the differentially expressed genes (DEGs). Functional enrichment analysis was performed, and protein-protein interaction (PPI) network was generated by the STRING (11.0) database and Cytoscape (3.7.2) software. Hub genes were screened and verified through GEPIA2 and GEO microarray database. Sensitivity of AML cell lines with high expression of hub genes to the small-molecule drugs were identified using GSCA Lite. Results A total of 456 DEGs were identified and top 100 genes were screened out, of which six genes (FLT3, PF4, CD163, MRC1, CSF2RB, PPBP) were upregulated in AML and individually had a worse prognosis by the overall survival (OS) analysis. AML cell lines with FLT3-overexpression and CSF2RB-overexpression were sensitive to most small-molecule drugs, while, AML cells with CD163-overexpression were only sensitive to a few drugs. However, sensitivity to Erlotinib was correlated with high expression of PF4 and PPBP. Conclusions In summary, FLT3, PF4, CD163, MRC1, CSF2RB, PPBP may be potential biomarkers and potential sensitive small-molecule drugs were correlated with overexpression of the biomarkers in AML.

Nasal-type NK/T-cell lymphoma (nasal-type NKTL) is one of the most lethal cancers for. Our study aimed to identify hub differentially expressed genes (DE- genes) and their upstream microRNAs between nasal-type NK/T-cell lymphoma (NKTL) tumor samples and normal nasal tissues through integrated bioinformatics. The 503 DE-genes and 106 DE-miRNAs were identified between NKTL and human normal nasal samples. GO and KEGG analysis were significantly enriched in meiotic recombination, regulation of syncytium formation by plasma membrane fusion, deubiquitination, enriched in meiotic recombination, regulation of syncytium formation by plasma membrane fusion, and stem cell division. And 11 differential expression hub genes and their upstream microRNAs were identified between nasal-type NKTL and normal nasal samples. In summary, after a series of analyses, we found that 11 hub DE-genes and their upstream DE-miRNAs (CDC27- miR-548c-3p, FREM2- miR-373*, ARHGAP29-miR-548c-3p, QSER1-miR-548c-3p, CD3EAP-miR-149*, SF3A1- miR-548c-3p, AQP4-miR-29b, ZFP36L2-miR-142-3p, SRP72-miR-16, TSC22D2-miR-16, TSC22D2-let-7f, DOCK5-miR-16) between nasal-type NKTL and normal nasal samples. They are highly likely to be serve as promising biomarkers in nasal-type NKTL.

Long non-coding RNA plasmacytoma variant translocation 1 (PVT1) has been reported to be associated with oncogenesis. However, the functional role of PVT1 in Burkitt lymphoma has not yet been addressed. The purpose of the present study was to investigate the effect of PVT1 knockdown by small interfering RNA (siRNA) on the proliferation of Burkitt lymphoma Raji cells and to explore its possible mechanism of action. An effective siRNA targeting PVT1 was screened and the corresponding short hairpin RNA (shRNA) was reconstructed into a lentiviral vector. Cell proliferation and cell cycle distribution were assessed by Cell Counting kit-8 assay and flow cytometry, respectively. Protein expression levels of c-Myc, cyclin-dependent kinase inhibitor1A (CDKN1A, P21) and cyclin E1 (CCNE1) were detected by western blotting. A polymerase chain reaction (PCR) array was used to analyse the expression of genes associated with the cell cycle. PVT1 knockdown markedly suppressed proliferation, and induced cell cycle arrest at the G0/G1 phase in Raji cells. Protein expression levels of c-Myc and CCNE1 were reduced, whereas P21 protein expression was markedly increased following downregulation of PVT1 in Raji cells. The cell cycle PCR array revealed that 54 genes were upregulated and 26 genes were downregulated in Raji cells following PVT1 knockdown. Reverse transcription-quantitative PCR demonstrated that cyclin G2 (CCNG2), CDKN1A, Retinoblastoma-like 2 (RBL2, p130), HUS1 checkpoint homolog, cyclin dependent kinase inhibitor 3 (CDKN3) and cyclin dependent kinase inhibitor 1B (CDKN1B) expression were upregulated, whereas the expression levels of CCNE1, cyclin D1 (CCND1) and cell division cycle 20 (CDC20) were downregulated in Raji cells with PVT1 knockdown. In conclusion, PVT1 knockdown may inhibit the proliferation of Raji cells by arresting cells in G0/G1 phase. Furthermore, inhibition of cell proliferation may be associated with a reduction inc-Myc expression and alterations in the expression levels of cell cycle-associated genes.

Background This study aimed to investigate the effects of microRNA19a (miR-19a) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of acute myeloid leukemia cells (HL60). Methods In experiment 1, HL60 cells were divided into the blank control group, the blank transfection group, the scrambled oligonucleotide (SODN) group and the ASODN group. MiR-19a ASODN and SODN were independently transferred into HL60 cells. The miR-19a expression was detected by real-time quantitative RT-PCR (qRT-PCR) after 48-h and 72-h transfection; CCK8 assay was used to detect the proliferation inhibition rate at 48 and 72 h; Hoechst 33258 staining was performed to examine apoptotic cells at 48 h; the apoptosis rate was detected by flow cytometry after AnnexinV/PI staining at 48 and 72 h; the protein expression of E2F1 and Bim was detected by Western blotting at 48 h. In experiment 2, cells were divided into the Ara-C group, the SODN + Ara-C group and the ASODN + Ara-C group. The cell proliferation inhibition rate at 48 and 72 h and apoptosis rate at 72 h were assessed as mentioned above. Results MiR-19a expression in the miR-19a ASODN group was lower than in the SODN group and the blank control group (P<0.05). MiR-19a ASODN significantly inhibited the growth of HL60 cells (P<0.05) and increased their apoptosis, and the apoptosis rate peaked at 48 h. The protein expression of E2F1 and Bim in the ASODN group was higher than in the blank control group, blank transfection group and SODN group. In addition, Ara-C further inhibited the growth and induced the apoptosis of miR-19a ASODN-transfected cells (P<0.05) in a time dependent manner. The growth inhibition rate and apoptosis rate in the ASODN + Ara-C group were higher than the sum of those in both Ara-C group and ASODN group. Conclusions miRNA-19a ASODN can inhibit the proliferation and induce apoptosis of HL60 cells and may exert synergistic effects with Ara-C on HL60 cells.

Objective: The effects of microRNA-20a (miR-20a) antisense oligonucleotides (ASODNs) on the proliferation and apoptosis of K562 cells were investigated, and the effects of these ASODNs in combination with imatinib on K562 cells were preliminarily observed. Methods: miR-20a ASODNs and scrambled oligonucleotides (SODNs) were chemically synthesized, and the later was used as the control. miR-20a ASODNs were transfected into K562 cells using Lipofectamine 2000 transfection reagent, and the expression of miR-20a was detected using real-time quantitative RT-PCR (qRT-PCR). The CCK8 assay was performed to detect the inhibition of the cell growth rate. The cells were stained by Hoechst 33258 to detect apoptotic cell morphology. Annexin V/PI double staining was used to detect the cell apoptosis rate using flow cytometry. The protein expression levels of E2F1, P21, and Bim in the K562 cell line were detected using western blotting. Results: The qRT-PCR results showed that the expression level of miR-20a in K562 cells transfected with miR-20a ASODNs was lower than those in the normal control, SODN and blank transfection groups (p < 0.05). miR-20a ASODNs significantly inhibited the growth of K562 cells as compared to the controls (p < 0.05). The Hoechst staining results showed morphological changes, suggesting apoptosis. The cell apoptosis rates in the ASODN group was (13.9 ± 1.5)%, which was significantly higher than that in the normal control group (1.84 ± 0.21)%, blank transfection group (3.21 ± 0.32)%, and SODN group (3.72 ± 0.44)% (p < 0.05). The protein expression of E2F1 and P21 in K562 cells transfected with miR-20a ASODNs were higher, while the level of Bim protein was significantly lower than that in the control groups. When miR-20a ASODNs were combined with imatinib, the growth of K562 cells was significantly inhibited as compared to the ASODN treatment alone, imatinib alone, and SODN+imatinib groups (p < 0.05). Conclusions: miR-20a ASODNs could induce apoptosis and inhibit the proliferation of K562 cells. In addition, imatinib combined with miR-20a ASODNs can increase the inhibitory effect on K562 cell proliferation.

Objective To screen the effective small interfering RNAs (siRNAs) targeting against long non-coding RNA (lncRNA) PVT1 (plasmacytoma variant translocation 1), and to observe its influence on the proliferation of human leukemic cell K562.Methods Four siRNA sequences targeting against PVT1 (PVT1-siRNA1, 2, 3, 4) and scramble control sequence (SC-siRNA) were designed and synthesized.Using HiperFect transfection reagent, all of the siRNAs were transfected into K562 cells.The experiment was divided into siRNA1 group, siRNA2 group, siRNA3 group, siRNA4 group, SC-siRNA group, idling group and untransfected group.The PVT1 RNA expression level was assessed by real-time RT-PCR.The cell proliferation was determined by CCK8 assay and the cell cycle was detected by Flow cytometry.Results The expression level of PVT1 RNA of K562 cells in siRNA4 group was significantly reduced compared with SC-siRNA group, idling group and untransfected group 24 and 48 hours after transfection (P<0.01), while there was no significant difference among siRNA1 group, siRNA2 group, siRNA3 group and untransfected group (P>0.05).The cell proliferation of K562 cells in the siRNA4 group was significantly inhibited as compared with SC-siRNA group and idling group 24, 48 and 72 hours after transfection (P<0.01).The cells in the siRNA4 group were arrested at G1 phase 48 and 72 hours after transfection (P<0.01).Conclusion PVT1-siRNA can inhibit the proliferation of K562 cells through arresting the cell cycle at G1 phase.

AIM:Bcl11a is closely associated with B-cell lymphoma and B-cell chronic lymphocytic leukemia. However, little is known about the expression character of Bcl11a gene in B-cell acute lymphoblastic leukemia (B-ALL). In previous study, our results showed there was a clear dysregulation in the global gene expression of the Bcl11a-suppressed B lymphoma cells. The aim of the study is to further evaluate the role of Bcl11a and the expression level of its related genes Mdm2, Pten in B-ALL patients.METHODS:The relative mRNA expression levels of Bcl11a and Mdm2, Pten in blood mononuclear cells of B-ALL patients were determined by real-time quantitative reverse transcript-polymerase chain reaction (qRT-PCR) with SYBR Green Dye. Peripheral blood mononuclear cells from healthy volunteers served as control. Glyceraldehyde-3-phosphate dehydrogenase (Gapdh) was used as reference.RESULTS:There was higher Bcl11a mRNA expression in initial diagnosed B-ALL than the healthy control, but there is no statistical significance (P > 0.05). The expression level of Bcl11a and Mdm2 in initial diagnosed B-ALL patients was statistically higher compared to those with complete remission (CR; P < 0.05). The expression levels of Bcl11a and Mdm2, Pten in B-ALL patients with CR were decreased significantly when compared with the healthy control (P < 0.05). The correlation analyses revealed that the Bcl11a expression level was positively correlated with the Mdm2 and Pten relative expression level in B-ALL patients. There is also positive correlation between Mdm2 and Pten expression.CONCLUSIONS:Bcl11a transcript levels were significantly downregulated in B-ALL patients with CR. The decreased expression characteristics of Bcl11a and Mdm2, Pten might imply the CR of B-ALL patients.

Objective We aimed to evaluate the expression pattern of the genes BIM, BCL-6, and c-MYC in adult patients at initial diagnosis of B-cell acute lymphoblastic leukemia（B-ALL）.Methods Relative m RNA levels of BIM, BCL-6, and c-MYC in peripheral blood mononuclear cells（PBMCs） from B-ALL patients were determined by quantitative reverse-transcription polymerase chain reaction（q RT-PCR） using SYBR Green dye. PBMCs from healthy volunteers served as a control. GAPDH was used as a reference gene.Results Relative expression of BIM, BCL-6, and c-MYC m RNA in B-ALL patients was significantly lower than in healthy controls（P ＜ 0.05）. Furthermore, this result was observed for both newly diagnosed B-ALL patients and those incomplete remission（CR）（P ＜ 0.05）. There were no statistically significant differences in the expression levels of BIM, BCL-6, and c-MYC between these B-ALL patient groups（P ＞ 0.05）. Spearman’s rank correlation analyses revealed the expression level of BIM to be positively correlated with that of BCL-6 in B-ALL patients.Conclusion Expression of the genes BIM, BCL-6, and c-MYC is decreased in adult B-ALL patients. Moreover, the expression pattern of these genes may be similar in such patients at initial diagnosis and following CR. The expression characteristics of BIM, BCL-6, and c-MYC may constitute useful markers for the diagnosis of adult B-ALL.

Objective To study the influence of down-regulating long non-coding RNA(lncRNA)plasmacytoma variant translocation 1(PVT1) andMYC-induced long noncoding RNA(MINCR)on the proliferation of lymphoma cells Raji and the probable mechanism.Methods Small interfering RNA(siRNA)targeting against PVT1 and MINCR as well as scramble control siRNA(SC-siRNA)were designed and synthesized chemically.The experiment was divided into PVT1-siRNA group, MINCR-siRNA group, PVT1-siRNA+MINCR-siRNA group, scramble control group and untransfected cells group.Using HiperFect transfection reagent,all of the siRNAs were transfected into Raji cells.The MINCR RNA expression level was detected by real-time RT-PCR.The cell proliferation, the cell cycle and the expression of c-Myc protein were detected by CCK8 assay, Flow cytometry and Western blot, respectively.Results The expression level of MINCR RNA ofMINCR-siRNA group was significantly reduced compared with that of the scramble control group and untransfected cells group(P<0.05).The cell proliferation was significantly inhibited after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells (P<0.05),which was more obvious than that in PVT1-siRNA group and MINCR-siRNA group(P<0.05).Consistently,the cells were arrested in G1 phase after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells (P<0.05),and it was more obvious compared with PVT1-siRNA group and MINCR-siRNA group(P<0.05).The expression of c-Myc protein was significantly decreased after PVT1-siRNA combining with MINCR-siRNA transfected into Raji cells(P<0.05),which was more obvious compared with PVT1-siRNA group and MINCR-siRNA group(P<0.05).Conclusion Down-regulating PVT1 and MINCR can enhance the inhibition of proliferation of Raji cells.

Objective To investigate the role of miR-155 in radio-resistance of lymphoma cell line Raji cells.Methods The expression of miR-155 was detected by quantitative real-time PCR, cell growth was detected by CCK-8 assay, cell apoptosis was detected by flow cytometry, and the protein expressions of PTEN and p-AKT were detected by Western blot.Results The Raji cells had high levels of miR-155 RNA and p-AKT protein had a very low level of PTEN protein.Cell apoptosis was induced by 4.0 Gy X-ray irradiation after 48 h.The transfection of cells with miR-155 siRNA not only silenced miR-155, but also significantly decreased p-AKT level, increased the expression of pten gene and protein level, inhibited cell proliferation, and increased the radiation sensitivity of Raji cell, the apoptosis rate of siRNA + irradiation group(36.78％ ± 1.35％) was higher than that of irradiation group (t =12.572, P ＜ 0.05).Conclusions High levels of miR-155 was involved in radiation resistance of lymphoma Raji cells and this activity is likely modulated by the PTEN/PI3K/AKT signaling pathway.

By investigating the anti-adipogenic effects of WEHI-3 cells - a murine acute myelomonocytic leukemia cell line - we sought to improve the efficiency of hematopoietic stem cell transplantation (HSCT). Analysis of Oil Red O staining and the expression of adipogenic genes, including PPARγ, C/EBPα, FAS and LPL, indicated that WEHI-3 cells significantly inhibited 3T3-L1 mouse preadipocyte cells from differentiating into adipocytes. In vivo, fat vacuoles in mice injected with WEHI-3 cells were also remarkably reduced in the murine bone marrow pimelosis model. Moreover, the key gene in the Rho signaling pathway, ROCKII, and the key gene in the Wnt signaling pathway, β-catenin, were both upregulated compared with the control group. siRNA-mediated knockdown of ROCKII and β-catenin reversed these WEHI-3-mediated anti-adipogenic effects. Taken together, these data suggest that WEHI-3 cells exert anti-adipogenic effects and that both ROCKII and β-catenin are involved in this process.

Objective The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II (BMPR2), E1A binding protein p300 (EP300), transforming growth factor-β2 (TGFβ2), and tumor necrosis factor, and alpha-induced protein 3 (TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcript-polymerase chain reaction (qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as reference. Results The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control (P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control (P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group (P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3 (r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control (P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines (P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells (P<0.05). Conclusion Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.

目的 设计靶向B细胞淋巴瘤/白血病11A基因的小干扰RNA(BCL1 1A-siRNA),并观察其对人胚胎肾细胞株HEK293细胞增殖的影响.方法 利用siRNA Target Finder工具,针对BCL11A设计合成siRNA319、siRNA585、siRNA2292、siRNA475.将HEK293细胞接种不含抗生素的培养基中,分为阳性对照组(PC组)、阴性对照组(NC组)、BCL11A-siRNA319组(si319组)、BCL11A-siRNA585组(si585组)、BCL11A-siRNA2292组(si2292组)、BCL11A-siR-NA475组(si475组)、空转组、细胞组.通过LipofectamineTM 2000,si319组将siRNA319、si585组将siRNA585、si2292组将siRNA2292、si475组将siRNA475分别转染HEK293细胞;转染后6h倒置荧光显微镜观察细胞转染情况并用流式细胞仪检测转染效率;转染后24、48、72 h,采用实时定量RT-PCR法观察BCL11A-siRNA对BCL11A基因表达的影响;CCK8法观察HEK293细胞体外增殖情况.结果 转染后24h,si585组、si2292组的HEK293细胞BCL11A基因表达降低,72 h逐渐恢复;si585组、si2292组细胞增殖受抑,与NC组、空转组、细胞组相比,P均＜0.05.结论 BCL11A-siRNA585及BCL11 A-siRNA2292可降低HEK293细胞中BCL1 1A基因表达,抑制细胞增殖.

Background: B cell chronic lymphocytic leukemia/lymphoma 11 A (BCL11A) is associated with human B cell malignancy initiation. Our previous study has shown that downregulation of BCL11A mRNA by small interfering RNA (siRNA) is capable of inducing apoptosis in the SUDHL6 cell line. To further explore the effects of BCL11A siRNA on the enhanced cytotoxicity of a chemotherapeutic drug, we investigated the effects of BCL11A siRNA combined with vincristine (VCR) on SUDHL6 cell proliferation and apoptosis.Methods: Chemically synthesized BCL11A siRNA was transfected into SUDHL6 cells using the HiPerFect Transfection Reagent in combination with VCR. Cell proliferation was measured by the CCK8 assay. The morphology of apoptotic cells was observed with Hoechst 33258 staining. The rate of cell apoptosis was determined by annexin V-fluorescein isothiocyanate/propidium iodide double staining using fluorescence-activated cell sorting (FACS) analysis.Results: After BCL11A siRNA plus VCR treatment, cell proliferation was significantly decreased in comparison with VCR or BCL11A siRNA treatment alone and negative control siRNA plus VCR treatment (P <0.05). The apoptotic rate of BCL11A siRNA plus VCR treated cells was significantly increased compared with BCL11A siRNA and VCR treatment alone and negative control siRNA plus VCR treatment (P <0.05).Conclusions: The combination of BCL11A siRNA and VCR increases apoptosis in SUDHL6 cells. Our study implies that BCL11A siRNA in combination with VCR may be a useful approach for improving effective treatment for B cell lymphoma.

Our previous study has shown that downregulation of B-cell chronic lymphocytic leukemia (CLL)/lymphoma11A ( BCL11A ) gene by small interfering RNA (siRNA) resulted in the growth inhibition and apoptosis of B cell lymphoma cell line SUDHL6. To gain further insight into the molecular mechanisms of this process and identify the differentially expressed genes in SUDHL6 cells after BCL11A downregulation, the global gene expression profile was identified and analyzed using the Affymetrix HG-U133 Plus 2.0 array. Twenty-one differentially expressed genes were validated and analyzed from the BCL11 A siRNA-treated SUDHL6 cells. There was a significant dysregulation in the global gene expression of the BCL11A -suppressed SUDHL6 cells. There were 1903 genes differentially expressed with >2-fold changes between the BCL11 A siRNA- and negative control-transfected cells. Of these, there were 916 upregulated genes and 987 downregulated genes. The differential genes are involved in various molecular functions and signaling pathways. QRT-PCR validation of the selected differentially expressed genes demonstrated there was a good correlation with the microarray analysis. There was a significant deregulation of expression in the apoptosis-related genes such as BCL-2 , BCL2L11 and involved in TGFβ, MAPK, WNT signaling pathways after BCL11A was downregulated in SUDHL6 cells. Our results show that the suppression of BCL11A by RNA interference altered gene expression profile of SUDHL6 cells. The apoptosis-related genes BCL-2 , BCL2L11 and the gene alterations in TGFβ, MAPK, WNT signaling pathways might be important in BCL11A siRNA-induced apoptosis of SUDHL6 cells, suggesting BCL11A is involved in gene networks associated with apoptosis.

[ABSTRACT]AIM:ToinvestigatetheeffectofmiR-155-specificsiRNAaloneorincombinationwithcytosinear-abinoside (Ara-C) on the growth and apoptosis of Burkitt lymphoma Raji cells .METHODS: miR-155-specific siRNA and/or Ara-C were used to treat the cells .Quantitative real-time polymerase chain reaction was used to detect the expres-sion of miR-155.The growth of the cells was analyzed by CKK-8 assay.The cell apoptosis was determined by flow cytome-try.RESULTS:The miR-155 expression level of the cells transfected with miR-155 siRNA was significantly lower than that in the 2 control groups .Ara-C or miR-155 siRNA alone inhibited the growth of Raji cells in a dose-depend manner . miR-155 siRNA combined with Ara-C produced more inhibition of cell proliferation (P<0.05).After treatment for 48 h, the apoptotic rate of Raji cells in miR-155 siRNA+Ara-C group [(38.4 ±1.4)%] was higher than that in Ara-C group [(16.5 ±0.3)%] and miR-155 siRNA group [(14.6 ±0.3)%], with statistically significant difference (P<0.05). The expression of caspase-3 in Ara-C+miR-155 siRNA group was increased significantly as compared with Ara-C group and miR-155 siRNA group.CONCLUSION:miR-155-specific siRNA enhances the chemosensitivity of Raji cells to Ara-C by inducing apoptosis through the caspase-3 pathway .

Our previous study has shown that downregulation of B‐cell chronic lymphocytic leukemia (CLL)/lymphoma11A ( BCL11A ) gene by small interfering RNA (siRNA) resulted in the growth inhibition and apoptosis of B cell lymphoma cell line SUDHL6. To gain further insight into the molecular mechanisms of this process and identify the differentially expressed genes in SUDHL6 cells after BCL11A downregulation, the global gene expression profile was identified and analyzed using the Affymetrix HG‐U133 Plus 2.0 array. Twenty‐one differentially expressed genes were validated and analyzed from the BCL11 A siRNA‐treated SUDHL6 cells. There was a significant dysregulation in the global gene expression of the BCL11A ‐suppressed SUDHL6 cells. There were 1903 genes differentially expressed with >2‐fold changes between the BCL11 A siRNA‐ and negative control‐transfected cells. Of these, there were 916 upregulated genes and 987 downregulated genes. The differential genes are involved in various molecular functions and signaling pathways. QRT‐PCR validation of the selected differentially expressed genes demonstrated there was a good correlation with the microarray analysis. There was a significant deregulation of expression in the apoptosis‐related genes such as BCL‐2 , BCL2L11 and involved in TGFβ, MAPK, WNT signaling pathways after BCL11A was downregulated in SUDHL6 cells. Our results show that the suppression of BCL11A by RNA interference altered gene expression profile of SUDHL6 cells. The apoptosis‐related genes BCL‐2 , BCL2L11 and the gene alterations in TGFβ, MAPK, WNT signaling pathways might be important in BCL11A siRNA‐induced apoptosis of SUDHL6 cells, suggesting BCL11A is involved in gene networks associated with apoptosis.

OBJECTIVE:B-cell lymphoma 2 (Bcl-2) is an important member of the Bcl-2 family of proteins that regulate the induction of apoptosis. This study aims to investigate whether Bcl-2 small interfering RNA (siRNA) combined with miR-15a oligonucleotides (ODN) could enhance methotrexate (MTX)-induced apoptosis in Raji cells.METHODS:Chemically synthesized miR-15a ODN and Bcl-2 siRNA were transfected in Raji cells by using a HiPerFect Transfection Reagent and then combined with MTX. Expression levels of Bcl-2 protein were detected by Western blot. Cell proliferation was determined by CCK8 assay. The rate of cell apoptosis was determined by Annexin V/PI double staining. The morphology of apoptotic cells was observed by Hoechst-33 258 staining.RESULTS:After the cells were transfected with miR-15a ODN combined with Bcl-2 siRNA, Bcl-2 protein levels were evidently decreased. CCK8 assay showed that cell proliferation was significantly decreased and was significantly lower in miR-15a ODN combined with Bcl-2 siRNA plus MTX group than in miR-15a ODN with methotrexate group, Bcl-2 siRNA with MTX group, and single MTX group (P<0.05). Hoechst 33258 staining revealed numerous apoptotic cells. AnnexinV/PI double staining showed that the apoptotic rates were (13.13±1.60)%, (34.47±2.96)%, (32.87±3.48)%, and (45.47±2.16)% in MTX, Bcl-2 siRNA plus MTX, miR-15a ODN plus MTX, and miR-15a ODN combined with Bcl-2 siRNA plus MTX groups, respectively. Among these groups, the apoptotic rate of miR-15a ODN combined with Bcl-2 siRNA plus MTX group was the highest; this apoptotic rate was also significantly different from that of miR-15a ODN or Bcl-2 siRNA plus MTX (P<0.05).CONCLUSIONS:Bcl-2 siRNA combined with miR-15a ODN could enhance MTX-induced apoptosis in Raji cells. Bcl-2 siRNA and miR-15a combined with MTX may be a useful approach to improve the treatment effects on lymphoma.