Expression of connexin 32 and connexin 43 in acute myeloid leukemia and their roles in proliferation
Expression of connexin 32 and connexin 43 in acute myeloid leukemia and their roles in proliferation
复制标题
急性髓系白血病中连接蛋白 32 和连接蛋白 43 的表达及其在增殖中的作用。
DOI：
10.3892/ol.2012.884
复制
发表时间：
2012-11-01
期刊：
影响因子：
2.9
通讯作者：
Cui, Guohui
中科院分区：
文献类型：
--
作者：
Yi, Sha;Chen, Yan;Cui, Guohui
连接蛋白（Cxs）是一个保守的跨膜蛋白家族，在多细胞生物中通过缝隙连接参与细胞间通讯。然而，Cxs在急性髓系白血病（AML）中的作用知之甚少。本研究探讨急性髓系白血病细胞增殖与连接蛋白43（Cx43）和连接蛋白32（Cx 32）mRNA和蛋白表达的关系。采用生长曲线法观察细胞增殖情况，MTT法检测细胞增殖率。流式细胞仪分析细胞周期和细胞增殖指数。RT-PCR检测差距连接基因Cx43和Cx 32的mRNA表达。Western blot和免疫荧光法检测细胞系中Cx43和Cx 32蛋白的表达。OCI-AML 3和OCIM 2的倍增时间分别为48 h和36 h。在OCIM 2中，S期细胞百分比为59.47± 9.6%，增殖率为78.12±8.9%;而在OCI-AML 3中，S期细胞百分比为24.95± 5.8%，增殖率为35.21± 6.7%。在mRNA水平上，两种细胞系均表达Cx43和Cx 32，且两种细胞系Cx43和Cx 32 mRNA表达无显著差异。在蛋白质水平上，Cx43的表达有显著差异，但Cx 32的表达无显著差异。OCIM 2细胞的增殖能力高于OCI-AML 3细胞，Cx43蛋白免疫印迹和荧光强度均高于OCI-AML 3细胞。结果提示，Cx43在AML细胞中的高表达可能对AML细胞的增殖能力起重要作用。
Connexins (Cxs), a conserved family of trans-membrane proteins, function in the organization of cell-cell communicatin via gap junctions in multicellular organisms. However, the role of Cxs in acute myeloid leukemia (AML) is poorly understood. In this study, we investigated the relationship between cell proliferation and expression of connexin 43 (Cx43) and connexin 32 (Cx32) mRNA and proteins in acute myeloid leukemia in vitro. Proliferation was observed using a growth curve and the rate of proliferation was detected by MTT assay in the acute myeloid leukemia cell lines OCI-AML3 and OCIM2. Cell cycle and cell proliferation index were assessed by flow cytometry analysis. The mRNA expression of the gap junction genes Cx43 and Cx32 was detected by RT-PCR. The expression of Cx43 and Cx32 proteins in the cell lines was analyzed by western blot analysis and immunofluorescence. The doubling time of OCI-AML3 and OCIM2 was 48 h and 36 h, respectively. In OCIM2, the percentage of cells in the S phase fraction was 59.47±9.6%, and the proliferation rate was 78.12±8.9%; however, in OCI-AML3, the percentage of cells in the S phase was 24.95±5.8%, and the proliferation rate was 35.21±6.7%. At the mRNA level, both cell lines expressed Cx43 and Cx32, and there was no significant difference in the expression of Cx43 and Cx32 mRNA in the two cell lines. At the protein level, there was a significant difference in the expression of Cx43, but not of Cx32. The proliferation ability of OCIM2 was higher than OCI-AML3, and OCIM2 exhibited higher Cx43 western blot and fluorescence intensities compared with OCI-AML3. The results suggest that a higher expression of Cx43 in AML cells may play a significant role in the proliferation ability.