Expressions of bcl-6, lpp and miR-28 genes in diffuse large B cell lymphoma cell lines / 中国实验血液学杂志

This study was purposed to explore the expressions of bcl-6, lpp and miR-28 genes in diffuse large B cell lymphoma (DLBCL) cell lines at the levels of gene and protein, and the relationship between them. Northern blot was used to detect bcl-6 and lpp mRNA expression in 8 DLBCL cell lines. Solution hybridization was used to measure miR-28 expression, and Western blot was performed for BCL-6 and LPP protein determinations. The results showed that the expression of bcl-6 mRNA was higher in the cell lines with Ig/BCL-6 translocation (Oc1-ly8, MD903, CTB-1, and MD901), and negative in those without Ig/BCL-6 translocation (HRC57 and K231). The expression of lpp mRNA in CTB-1 cell line was negative. MiR-28 was positive in all cell lines, and the expression levels from high to low were in line as follows: K231, CTB-1, MD903, HRC57, MD901, RCK8, OC1-LY8 and BEVA. BCL-6 protein was also positive in all of cell lines, and the expression levels from high to low were as follows: RCK8, BEVA, MD901, CTB-1, MD903, OC1-LY8, HRC57 and K231. LPP protein was negative in K231 cells, and the expression levels in other cells from high to low were line up as follows: HRC57, OC1-LY8, BEVA, RCK8, CTB-1, MD901 and MD903. The expression levels of bcl-6 and lpp mRNA were not consistent with expression levels of protein. It is concluded that the gene expression levels of bcl-6, lpp and miR-28 are different in various DLBCL cell lines. The expression levels of bcl-6 and lpp mRNA are not parallel with expression levels of protein. The roles of bcl-6, lpp and miR-28 in initiation and development of DLBCL need further investigation.

Lipoprotein lipase and serum thymidine kinase level in chronic lymphocytic leukemia and their correlations with other prognostic factors / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate lipoprotein lipase (LPL) and serum thymidine kinase (TK) levels in chronic lymphocytic leukemia (CLL) and their correlations with other prognostic factors.</p><p><b>METHODS</b>LPL expression level in peripheral blood samples of 58 CLL patients was detected by semi-quantitative reverse transcription PCR (RT-PCR). Serum TK1 level in 39 CLL patients was detected by enhanced chemiluminescence (ECL) and TK monoclonal antibody (Anti-TK mAb). IgVH mutation status was detected by multiplex PCR and sequencing of purified PCR products. The expression of ZAP-70 protein and CD38 were determined by flow cytometry . Panel probes and fluorescence in situ hybridization (FISH) were used to detect cytogenetic aberrations.</p><p><b>RESULTS</b>The median LPL expression level in CLL was 0.26 (0 -6.29), while undetectable in normal controls. LPL expression level was significantly correlated with IgVH mutation status, Binet stages, CD38 and cytogenetic aberrations. Patients with unmutated IgVH genes had higher LPL than those with IgVH mutations (P = 0.010). Patients in Binet stage B and C had higher LPL than those in stage A (P = 0.011). LPL level was higher in patients with CD38 > or = 30% (P = 0.001). Higher LPL level was found in patients with unfavorable cytogenetic aberrations (deletion in 17p13 or 11q22) than those with favorable cytogenetics (deletion in 13q as the sole abnormality) (P = 0.002). LPL level was not significantly correlated with sex, age, and ZAP-70 protein (P > 0.05). The level of TK1 was higher in CLL patients than that in normal control (P < 0.05). Patients with higher level of absolute lymphocyte count (ALC), lactate dehydrogenase (LDH), unmutated IgVH genes and ZAP-70 had higher levels of TK1 than those with lower level of ALC, LDH, mutated IgVH genes and ZAP-70 (P = 0.018, P = 0.018, P = 0.030 and P = 0.038, respectively). TK1 level had no correlation with sex, age, Binet stages, CD38, and cytogenetic aberrations (P > 0.05).</p><p><b>CONCLUSIONS</b>LPL expression and serum TK1 levels significantly correlate with other CLL prognostic factors and could be predictive markers for IgVH mutation status. LPL and serum TK1 might be applied to the assessment of prognosis in CLL patients.</p>

Prognostic significance of p53 and ATM gene deletion in patients with chronic lymphocytic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the prognostic significance of p53 and ATM gene deletion in patients with chronic lymphocytic leukemia (CLL).</p><p><b>METHODS</b>Interphase fluorescence in situ hybridization (FISH) and probes of LSI p53 and LSI ATM were used to detect p53 and ATM gene deletion in 80 patients with CLL. p53 and ATM gene deletion and their association with some prognostic factors were analyzed. The Kaplan-Meier method was used to calculate survivals, and results were compared using the Log-rank test. Multivariate COX regression analysis was used to assess associations between survival and potential risk factors.</p><p><b>RESULTS</b>Out of the 80 CLL patients, p53 gene deletion was found in 14 (17.5%) cases, ATM gene deletion in 9 (11.3% ) cases, and both the two genes deletion in 3 (3.8%) cases. There was no significant differences of p53 and ATM gene deletion rates in sex, age, Binet stages, peripheral lymphocyte count, and the levels of LDH, beta2-MG, and ZAP-70. The p53 and ATM gene deletion rates were higher in the group of CD38 high expression than that in the group of low expression (P=0.025 and P=0.001). Among 41 patients who received fludarabine containing protocol, none of the nine cases with p53 and/or ATM gene deletion achieved complete response (CR), while 12 of 32 (37.5%) cases without the gene deletion achieved CR. The survival time was shorter in patients with p53 gene deletion (P<0.01). There was no association between outcome and ATM gene deletion (P=0.556). On multivariate analysis by COX regression, p53 gene deletion and CD38 expression (P=0.014 and P=0.017, respectively) were found to be independent factors in predicting survival.</p><p><b>CONCLUSION</b>CLL patients with p53 and/or ATM gene deletion had poor therapeutic effect, and hence poor prognosis.</p>

DNA sequencing of miR-17-92 cluster at chromosome 13q31-q32 in mantel cell lymphoma cell lines / 中国实验血液学杂志

This study was aimed to explore the characteristics of miR-17-92 cluster at chromosome 13q31-q32 in B cell malignant lymphoma and to investigate the changes of miR-17-92 cluster in B cell lymphoma at genome DNA level and its influence on expression of miR-17-92 cluster and relation with lymphoma occurrence. PCR and DNA sequencing were used to detect miR-17-92 cluster at chromosome 13q31-q32 in mantel cell lymphoma (MCL) cell lines Rec1, G519 and Z138. The results showed that DNA sequence of miR-17-92 cluster at chromosome 13q31-q32 in MCL cell lines were normal. It is concluded that there is no abnormal change in DNA sequence of miR-17-92 cluster which is not the mechanism for miR-17-92 cluster overexpression in mantel cell lymphoma cell lines.

Prognostic significance of lactate dehydrogenase and beta2-microglobulin in chronic lymphocytic leukemia / 中国实验血液学杂志

To evaluate the prognostic value of lactate dehydrogenase (LDH) and beta2-microglobulin (beta2-MG) in chronic lymphocytic leukemia (CLL), a total of 141 cases of CLL had been investigated retrospectively. The Kaplan-Meier method was used to estimate the overall and failure-free survival distributions. Cox regression was used in univariate and multivariate analysis of potential predictors for overall survival. In multivariate analysis, the expression levels of LDH and beta2-MG were divided into 3 groups: (1) elevation of both LDH and beta2-MG levels; (2) elevation of LDH or beta2-MG levels alone; (3) normal levels of both LDH and beta2-MG. The results showed that serum LDH and beta2-MG levels of patients in Binet C were significantly higher than those in Binet A (p=0.034 and p=0.035). The level of serum beta2-MG was not correlated with lymphocyte count (p=0.756). Binet C and high LDH level were associated with significantly shorter overall survival. beta2-MG was not proved to have any association with overall survival. The overall survival time in group of elevation of both LDH and beta2-MG levels was shorter than that in group of normal levels of both LDH and beta2-MG. It is concluded that serum LDH level and Binet stage are important prognostic factors for CLL.