Effect of miR-202 on the growth of multiple myeloma cells via regulating B cell-activating factor and the underlying mechanism / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the regulating effect of miR-202 on B cell-activating factor, and check whether the regulation influences the growth of multiple myeloma cells.</p><p><b>METHODS</b>The potential binding sites of BAFF for miR-202 were predicted using bioinformatics software. Luciferase reporter gene analysis was used to evaluate the regulatory effect of miR-202 on BAFF. Human multiple myeloma U266 cells were transfected with has-miR-202-mimics, has-miR-202-inhibitor, siBAFF and their negative controls, respectively. After above treatments, BAFF mRNA and protein levels were detected by real-time PCR and Western blot analysis, and the proliferation and apoptosis in the multiple myeloma (MM) cells were examined by WST-1 and annexin V-FLUOS assay, respectively.</p><p><b>RESULTS</b>The BAFF mRNA expression levels in the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.040 ± 0.057, 0.573 ± 0.073, 1.205 ± 0.097 and 0.368 ± 0.052, respectively. BAFF mRNA expressions in U266 cells transfected with has-miR-202-3P-mimics and siBAFF were significantly decreased compared with that in the untransfected group (P < 0.05). The BAFF protein expression level of each group was consistent with the mRNA assay result. The absorbance value in 450 nm of the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.063 ± 0.052, 0.714 ± 0.045, 0.936 ± 0.066 and 0.764 ± 0.053, respectively. In comparison with the untransfected group, the absorbance value at 450 nm of has-miR-202-3P-mimics and siBAFF transfected groups was significantly reduced (P < 0.05). The cell apoptosis rates of untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 26.2%, 49.6%, 21.1% and 30.7%, respectively. Therefore, the cell apoptosis rate of has-miR-202-3P-mimics transfected group was significantly increased than that of the untransfected group (P < 0.05). p-JNK protein expression level was decreased in the has-miR-202-3P-mimics transfected cells.</p><p><b>CONCLUSIONS</b>MiR-202 can inhibit the proliferation and induce apoptosis in MM cells via regulating BAFF. JNK/SAPK signaling pathway is involved in the regulation of BAFF by miR-202.</p>

Abnormal expression of APRIL in colorectal cancer cells promotes tumor growth and metastasis / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the effects of a proliferation-inducing ligand (APRlL) on colorectal cancer (CRC) cell growth and migration, and to observe the role of APRIL in CRC biological behavior.</p><p><b>METHODS</b>The siRNA plasmid vector targeting APRIL gene (APRIL-siRNA) was transfected into human colorectal cancer SW480 cells and recombinant human APRIL (rhAPRIL) was used to stimulate human colorectal cancer HCT-116 cells. Cell proliferation activity was analyzed using cell counting kit-8 (CCK-8), cell cycle was detected by flow cytometry, and the protein expression of cyclin D1, p21 and Bcl-2 was detected by Western blot analysis. Tumor cell migration and invasion were measured by Transwell chambers. RT-PCR was applied to examine the mRNA expression level of MMP-2 and MMP-9. APRIL-siRNA was used to transfect directly SW480 cells, which were injected subcutaneously into nude mice, then the tumor growth and metastasis were observed.</p><p><b>RESULTS</b>Cell proliferation ability of APRIL-siRNA-transfected SW480 cells was drastically repressed, and the percentage of G0/G1 phase cells was significantly increased (t = 4.12, P < 0.05), accompanied with depressed cyclin D1, Bcl-2 expression and elevated p21 expression. Cell proliferation ability of rhAPRIL-stimulated HCT-116 cells was promoted with a decreased G0/G1 phase ratio (t = 3.31, P < 0.05). cyclin D1 and Bcl-2 protein expression was up-regulated while p21 was down-regulated by rhAPRIL stimulation. Metastatic and invasive capacities of APRIL-siRNA-transfected SW480 cells were significantly inhibited compared with their respective controls (both P < 0.05), accompanied with the deregulated MMP-2 and MMP-9 mRNA expression. Metastatic and invasive capacities of rhAPRIL-stimulated HCT-116 cells were promoted with up-regulated MMP-2 and MMP-9 mRNA expression(both P < 0.05). Tumor growth in the group transfected with APRIL-siRNA appeared to be slower than that in the control groups and the expression of MMP-2, MMP-9 in tumor tissues was depressed in the APRIL-siRNA group.</p><p><b>CONCLUSIONS</b>APRIL facilitates tumor growth and metastasis, and is associated with carcinogenesis and prognosis. Our findings suggest that APRIL might be used as a novel target for the intervention and therapy of colorectal cancer.</p>

Expression of B lymphocyte stimulator and its receptors in multiple myeloma cells: a mechanism for the cell growth and survival / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of B lymphocyte stimulator (BlyS) and its receptors in multiple myeloma (MM) cells, and to explore the relationship between BLyS and the development of human multiple myeloma.</p><p><b>METHODS</b>Flow cytometry, RT-PCR and Western blot were used to examine the expression of BLyS and its receptors in MM (KM3 and CZ1) cells. Fluorescence immunocytochemical method and confocal laser scanning technique were applied to the localization of BLyS in KM3 cell. WST proliferation assay was used to examine the effect of BLyS on MM cells growth and survival. Linear correlation analysis was used to detect LDH and beta 2-microglobulin (beta2M) levels with BLyS protein and mRNA expressions in MM patients.</p><p><b>RESULTS</b>(1) BLyS and its receptors were expressed in MM cells. (2) BLyS protein was localized on the KM3 plasma membrane. (3) BLyS promoted survival and proliferation of MM cells. (4) MM patients had significantly higher expression levels of BLyS [77.42% (24/31)] BLyS mRNA [93.55% (29/31)], which were significantly correlated with the levels of LDH and beta 2-microglobulin (beta2M).</p><p><b>CONCLUSION</b>BLyS and its receptors in MM cell lines and MM patient bone marrow might have a potential role in the growth and survival of malignant plasma cells.</p>

Measurement of B-lymphocyte stimulator mRNA expression in children with infectious mononucleosis by real-time fluorescence quantitative method / 中国当代儿科杂志

<p><b>OBJECTIVE</b>B cell multiplication plays a key role in infections mononucleosis. The present study was designed to detect the expression of B-lymphocyte stimulator (BLyS) mRNA in peripheral blood using real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) in children with infectious mononucleosis in order to explore the role of BLys in this disorder.</p><p><b>METHODS</b>Specific primers and TaqMan probes of BLyS were designed, and fluorescence of the PCR products were detected continuously during amplification. According to the standard curves created by plasmid DNA, the expression level of target genes in clinical samples were calculated using Stata Software version 8.0, and the results were presented as the ratio of copies of target gene mRNA to beta2 microglobulin (beta2M) mRNA copies. BLyS mRNA expression in peripheral blood was measured by RFQ-PCR in 18 children with infectious mononucleosis and the results were compared with those measured in 15 healthy controls.</p><p><b>RESULTS</b>The range of target gene mRNA detected by REQ-PCR was from 109 ng/L to 101 ng/L. The coefficient of variation for intra-experimental and inter-experimental reproducibility ranged from 1.88% to 5.89% and 6.32% to 12.34%, respectively. BLyS mRNA expression in peripheral blood in children with infectious mononucleosis were significantly higher than that in controls (1.65+/-0.10 vs 0.56+/-0.08; P < 0.01).</p><p><b>CONCLUSIONS</b>RFQ-PCR has a high sensitivity and reproducibility for the measurement of BLyS mRNA expression. BLyS may be involved in the development of infectious mononucleosis.</p>