弥漫大B细胞淋巴瘤细胞系中bcl-6、lpp和miR-28基因表达的研究

Expressions of bcl-6,lpp and miR-28 Genes in Diffuse Large B Cell Lymphoma Cell Lines

本研究从基因和蛋白水平探讨弥漫大B细胞淋巴瘤(DLBCL)细胞系中Bcl-6、LPP和miR-28表达,及其之间的相互关系。应用Northern blot检测8个DLBCL细胞系(CTB-1、MD901、OC1-LY8、BEVA、RCK8、MD903、HRC57和K231)bcl-6和lpp的mRNA水平,液相杂交法检测miR-28表达,Western blot检测BCL-6和LPP的蛋白水平。结果显示,bcl-6基因易位类型涉及Ig基因细胞系(Oc1-ly8、MD903、CTB-1和MD901),其bcl-6mRNA的表达较强,而在未涉及Ig基因易位的细胞系(HRC57和K231)中未见表达。lpp mRNA在CTB-1细胞系中表达阴性。miR-28在各细胞系中均有表达,表达强度高到低依次为:K231、CTB-1、MD903、HRC57、MD901、RCK8、OC1-LY8和BEVA。BCL-6蛋白在各细胞系表达由强到弱依次为:RCK8、BEVA、MD901、CTB-1、MD903、OC1-LY8、HRC57和K231;而LPP蛋白在K231细胞系中未见表达,余各细胞系表达由强到弱依次为:HRC57、OC1-LY8、BEVA、RCK8、CTB-1、MD901和MD903。各细胞系中bcl-6和lpp mRNA水平与蛋白的表达并不一致,即高mRNA水平并不一定导致蛋白的表达增加。结论:不同DLBCL细胞系具有不同的BCL-6、LPP和miR-28蛋白表达水平,各细胞系中bcl-6和lpp mRNA表达水平与它们表达的蛋白量不平行,有关bcl-6、lpp和miR-28基因在DLBCL发生、发展中的作用机制有待进一步探讨。

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： RCKS, 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.