Clinical significance of serum free light chain in patients with multiple myeloma / 中国实验血液学杂志

Multiple myeloma (MM) is a malignant disorder characterized by the proliferation of a single clone of plasma cells that can produce excessive amounts of serum free light chain (sFLC). sFLC plays an important role in MM diagnosis and disease monitoring. The quantitative immuno-nephelometric assay is sensitive and specific means for sFLC testing. The aim of this study was to investigate the levels of sFLC in multiple myeloma and the relationship between sFLC and serum total light chain (sTLC). sFLC in 45 newly diagnosed patients were detected by immuno-nephelometric assay, and then the ratio of free kappa to free lambda for every sample was calculated. Meanwhile, sTLC was also determined in these patients. The results showed that the difference of sFLC levels between MM patients and the normal controls was significant (tΚ = 8.86, P < 0.001; tλ = 15.48, P < 0.001;tΚ/λ = 5.54,P < 0.005). No correlation between sFLC and sTLC was found in MM patients. It is concluded that the level of sFLC in MM patients is significantly higher than that in normal controls. sFLC and its ratio may be served as a indicator for diagnosis of MM. sTLC can not replace the role of sFLC.

Expression level of microRNA-92a and its clinical significance in multiple myeloma patients / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between the expression level of microRNA 92a (miR-92a) and del(13q14) and the prognosis of MM patients, and to explore the pathway that miR-92a involved.</p><p><b>METHODS</b>Bone marrow samples from 53 newly diagnosed MM patients were collected, del(13q14) was analyzed by interphase fluorescence in situ hybridization in sorted CD138 positive plasma cell. The expression of miR-92a in plasma cells was measured by quantitative real-time PCR. The expression of c-jun was detected by Western blot in miR-92a transfected MM cell lines (LP-1, U266 and JJN3).</p><p><b>RESULTS</b>Of the 53 MM patients, del(13q14) was detected in 31 (58.4%) patients. The median levels of miR-92a in MM patients with or without del(13q14) were 27.36±2.61 and 21.87±15.98, respectively (P>0.05). With the median follow-up of 13.5 (0.5-72.5) months, the median duration of progression-free survival of patients with high expression level of miR-92a was significantly shorter than those with low expression level of miR-92a (4.5 months vs 14.0 months, P=0.006). Overexpression of miR-92a in MM cell lines induces time-dependent down-regulation of c-jun.</p><p><b>CONCLUSIONS</b>High expression of miR-92a was associated with poor prognosis in MM patients. The expression level of miR-92a was not associated with del(13q14), and the effect of miR-92a on the progress of MM might be involved in c-jun pathway.</p>

Investigation of 1q21 amplification in patients with multiple myeloma using I-FISH and cIg-FISH / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the prevlance of 1q21 amplification in patients with multiple myeloma (MM) and its correlation with the progression and prognosis of the disease.</p><p><b>METHODS</b>1q21 amplification was detected in 48 patients with MM using cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization analysis (cIg-FISH) and interphase fluorescence in situ hybridization (I-FISH) analysis combined with CD138 immunomagnetic cell sorting (MACS).</p><p><b>RESULTS</b>1q21 amplification (≥ 3 red signals) was detected in 26/48(54.2%) cases by cIg-FISH and 31/48 (64.6%) cases by I-FISH combined with CD138 MACS. There was a good consistency between the two methods (P>0.05). The mortality of patients with 1q21 amplification was significantly higher than those without (P< 0.05). No significant difference was detected in terms of sex, age, Durie-Salmon stage, subgroup and international staging system (ISS) stage between patients with 1q21 amplification and those without (P>0.05).</p><p><b>CONCLUSION</b>The frequency of 1q21 amplification in MM is high. There was also an association between the amplification and poor prognosis. cIg-FISH is consistent with CD138 MACS combined with I-FISH.</p>

Detection of complex chromosomal aberrations in patients with multiple myeloma using multiplex fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To explore the value of multiplex fluorescence in situ hybridization (M-FISH) in the detection of the complex chromosomal aberrations (CCAs) in multiple myeloma (MM).</p><p><b>METHODS</b>M-FISH was used in 10 MM patients with CCAs detected by conventional cytogenetics (CC) using R-banding to refine the rearrangement of CCAs and identify the characteristics of marker chromosome.</p><p><b>RESULTS</b>M-FISH confirmed the 29 structural aberrations shown by CC analysis, and also confirmed the specific source of 21 types of chromosomal aberration, which were not detected by CC analysis. Among them, t(2;15)(q33;q22), t(6;7)(q23;q34), t(8;11) (q24;q23), t(1;14)(q10;q32) and t(X;1)(q26;q25) were new chromosomal aberrations. The median survival time of 9 MM patients with CCAs was 23 months and evidently shorter than that of MM patients without CCAs, with the mean survival time being 34 months.</p><p><b>CONCLUSION</b>M-FISH could refine CCAs in MM patients, find or correct the missed or misidentified abnormalities analyzed by CC. It has provided one of the essential methods for the research of chromosomal aberrations in MM.</p>

Investigation of the molecular changes in patients with multiple myeloma by fluorescence in situ hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the incidence and prognosis of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in patients with multiple myeloma (MM).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (I-FISH) with four different specific probes for the regions containing 1q21, 13q14.3 (D13S319), 14q32 and TP53 gene were performed in 43 MM patients.</p><p><b>RESULTS</b>Among the 43 MM patients, 1q21 amplification was observed in 28 (65.1%) cases, 13q14 deletion in 30 (69.7%) cases, TP53 gene deletion in 8 (18.6%) cases, and IgH translocation in 29 (67.4%) cases. The mortality of MM patients with 1q21 amplification, 13q14 deletion or TP53 gene deletion was higher than those without them.</p><p><b>CONCLUSION</b>There is high frequency of 1q21 amplification, 13q14 deletion, TP53 gene deletion and IgH translocation in multiple myeloma, and 1q21 amplification, 13q14 deletion and TP53 gene deletion are poor prognosis factors for MM patients.</p>

Investigation of chromosome 1 aberrations in patients with multiple myeloma using cIg-FISH method and its significance / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the incidence of 1q21 amplification and 1p12 deletion, and analyze the correlation between these aberrations with disease progression, prognosis and outcome in patients with multiple myeloma (MM).</p><p><b>METHODS</b>Cytoplasm light chain immunofluorescence with simultaneous interphase fluorescence in situ hybridization (cIg-FISH) was used to detecte the 1q21 amplification and 1p12 deletion in 48 patients with MM.</p><p><b>RESULTS</b>1q21 amplification (≥ 3 red signals) was determined in 26 of 48(54.2%) cases. The mortality of patients with 1q21 amplification was significantly higher than that of those lacking 1q21 amplification (P < 0.05). The sex, age, D-S stage, subgroup and ISS stage between patients with and without 1q21 amplification had no significant difference (P > 0.05). There was a significant difference in D-S stage and mortality between patients with 3 and with 4 copies of 1q21 (P < 0.05). No significant difference in sex, age, subgroup, ISS stage, and isotype was found between them (P > 0.05). 1p12 deletion (< 2 green signals) was found in 14 of 48 (29.2%) cases. There was no significant difference in sex, age, D-S stage, ISS stage, isotype, subgroup, and mortality between patients with and without 1p12 deletion.</p><p><b>CONCLUSION</b>The frequency of chromosome 1 aberrations in multiple myeloma is high and 1q21 amplification is a poor prognosis factor.</p>

microRNA-21 and microRNA-30b expression in multiple myeloma / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression of miR-21 and miR-30b in multiple myeloma (MM).</p><p><b>METHODS</b>Peripheral blood mononuclear cells from patients with MM were cultured at 2.5 x 10(6) cells/ml in alpha-MEM supplemented with 10% of fetal bovine serum, antibiotics, RANKL (50 ng/ml), and macrophage colony-stimulating factor (25 ng/ml) for 10 to 14 days to obtain osteoclasts with bone-resorbing activity. Primary myeloma cells were purified from 12 MM patients. Of them, 8 samples were cocultured with osteoclasts and 4 as noncocultured control. The expression of miR-21 and miR-30b was detected by real-time PCR.</p><p><b>RESULTS</b>The viability of MM cells recovered from cocultures was higher than those of noncocultured control. After cocultured with osteoclasts, primary myeloma cells from eight patients exhibited a 1.3- 5.9-fold increase in miR-21 expression and 1.38- 4.32-fold decrease in miR-30b expression compared with controls. In highly purified plasma cells from 3 healthy subjects, 12 MM patients and 11 MM cell lines, the expression of miR-21 was 1.9 +/- 0.8, 6.5 +/- 4.9 and 35.1 +/- 36.2, respectively; the expression of miR-30b was 13.6 +/- 1.8, 7.2 +/- 6.3 and 4.5 +/- 1.9, respectively.</p><p><b>CONCLUSIONS</b>miR-21 acts as an oncogene and miR-30b a tumor suppressor gene in MM.</p>

Role of bone marrow microenvironment in regulation of AP-1 gene expression in multiple myeloma cells / 中国实验血液学杂志

This study was aimed to investigate the role of bone marrow microenvironment in the regulation of activator protein 1 (AP-1) expression in multiple myeloma (MM) cells. The primary myeloma cells (CD138(+) cells) from 8 patients with MM were sorted by using immunomagnetic beads and were cocultured with osteoclasts in alpha-MEM supplemented with 10% fetal bovine serum, antibiotics, RNAKL (50 ng/ml) and macrophage colony-stimulating factor (25 ng/ml) for 10 to 14 days at 2.5 x 10(6) cells/ml. The expression levels of genes c-jun, junD fos and fosB were detected by real-time PCR. The results showed that the osteoclasts were observed after coculture of mononuclear cells in peripheral blood of MM patients with osteoclasts for 10 - 14 days. As compared with control (without coculture with osteoclasts), the viability of MM cells cocultured with osteoclasts obviously increased, the expression levels of c-jun, junD, fos and fosB decreased to 25.7% - 1.66%, 68.49% - 8.54%, 10.35% - 0.19% and 36.63% - 3.44% of the control respectively. It is concluded that the bone marrow microenvironment can inhibit the expression of c-jun, junD, fos and fosB promote myeloma cell proliferation and maintain cell survival.

Fluorescence in situ hybridization on bone marrow smear in the detection of cytogenetic aberrations of multiple myeloma / 中国实验血液学杂志

This study was aimed to establish the technique of interphase fluorescence in situ hybridization (I-FISH) used on smear of bone marrow directly, and to develop a new method for detection of the molecular cytogenetics in multiple myeloma (MM). After a series of treatment, fixation and digestion of the bone marrow smear as the carrier, the chromosome 8 centromere probe were used in I-FISH for molecular cytogenetics detection. At the same time, differences were compared in the results between the new method and the conventional I-FISH. The results showed that there was no statistically significant difference of proportion of various signals in non-hematologic malignancies when detected with the two methods (p>0.05). In bone marrow smear I-FISH, 8 out of 19 cases (42.1%) had abnormality of chromosome 8, including 5 cases with -8 (26.3%) and 3 cases with +8 (15.8%). It is concluded that the I-FISH on smear of bone marrow is characterized by convenience, economy and accuracy. Therefore, it can be used for research of molecular cytogenetics in MM.

Correlation between chromosome 13q14 deletion and 1q abnormality in multiple myeloma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the correlation between chromosome 13q14 deletion [del(13q14)] and chromosome 1q abnormality in multiple myeloma (MM).</p><p><b>METHODS</b>The bone marrow plasma cells of 48 previously untreated MM patients were purified by CD138 and magnetic cell sorting system, and interphase fluorescence in situ hybridization (I-FISH) was applied to detect the del(13q14) with D13S319 probe and the abnormalities of chromosome 1q with CEP1 SpectrumOrange probe in sorted MM cells.</p><p><b>RESULTS</b>Among the 48 MM patients, del(13q14) was observed in 22(45.8%) cases, the abnormalities of chromosome 1q were observed in 23 (47.9%) cases, among which 2 were 1q deletion and 21 were 1q duplication. The chromosome 1q abnormality was detected in 16 of the 22 cases of MM with del(13q14) and in 7 of the 26 cases of MM without del(13q14), and there was significant difference between the two groups (chi-square was 10.02, P was less than 0.01).</p><p><b>CONCLUSION</b>There is high frequency of chromosome 13q14 deletion and 1q abnormality in multiple myeloma. The chromosome 1q abnormalities are highly associated with 13q14 deletion.</p>