ABSTRACT
Objective:To investigate the expression of LncRNA ANCR in human glioma tissues and its relationship with malignant proliferation of cells.Methods:The samples of 10 normal brain tissue,13 low-grade and 45 high-grade gliomas were regarded as normal group, low-grade group and high-grade group, which were collected from neurosurgery department in Linyi Central Hospital, and the expression of ANCR and potential interaction molecule eIF4B was detected by reverse transcription polymerase chain reaction (RT-PCR) in vitro. Lentivirus transfection in vitro was used to construct the U251 shRNA ANCR and control cell line in human high-grade gliomas as control, test 1 and test 2 group cells in the study. QPCR detect the expression level ANCR, eIF4B and Myc mRNA in cells. Western blot was used to detect the expression of eIF4B and c-Myc protein, CCK-8 assay was used to detect the relative proliferation ability of cells, and the colony formation assay was used to observe the change of cell clone formation. SPSS 21.0 was used for statistical analysis, analysis of variance was used for inter group comparison, and SNK-q pairwise comparison method was used for intra group comparison.Results:The expressions of ANCR mRNA in high-grade glioma tissues, low-grade gliomas and normal brain tissues were 0.710±0.125, 2.033±0.312 and 3.408±0.296. The expressions of eIF4B mRNA in high-grade glioma tissues, low-grade gliomas and normal brain tissues were 0.176±0.019, 0.268±0.022 and 0.426±0.028. The expression of ANCR and eIF4B in high-grade glioma tissues was higher than that in low-grade gliomas and normal brain tissues ( P<0.001). The expression of ANCR in low-grade glioma tissues was higher than that in normal brain tissues ( P=0.013). There was a significant positive correlation between the expression of ANCR and eIF4B in glioma tissues ( P<0.001) ; The expressions of ANCR mRNA in Control, test1 and test2 were 1.000±0.021, 0.202±0.057 and 0.300±0.016. The expressions of eIF4B mRNA were 1.000±0.078, 0.452±0.012 and 0.526±0.037, and the expressions of c-Myc mRNA were 1.000±0.053, 0.688±0.067 and 0.564±0.089. the expressions of ANCR, eIF4B and c-Myc mRNA and protein in test1 and test2 cells were significantly lower than those in the control group ( P<0.01) ; the proliferation of test1 and test2 groups were significantly decreased at 72h and 96h, and the ability of colony formation was significantly decreased ( P<0.001) . Conclusion:The expression of ANCR was significantly up-regulated in high-grade glioma tissues and positively correlated with the expression of eIF4B. Interference with ANCR in vitro could mediate the decrease of the expression of eIF4B and c-Myc mRNA and protein molecules, thereby inhibiting the proliferation of glioma cells.
ABSTRACT
Human acute leukemia (AL) is a clonal malignancy with abnormal hematopoietic stem cells. Clinically, AL is very difficult to cure due to its sudden onset and short course of disease progression. Previous studies have shown that eukaryotic initiation factor 4B (eIF4B) plays a critical role in the development of chronic leukemia. However, the involvement of eIF4B in human acute leukemia is still largely unknown. Therefore, we studied eIF4B function and its regulatory mechanism in human acute leukemia. We found that phosphorylation levels of eIF4B in acute leukemia cells were significantly reduced in response to treatment with either LY294002 (PI3K inhibitor), AKTi (AKT inhibitor) or SMI-4A (Pim inhibitor). Co-treatment with inhibitors targeting JAK/STAT5/Pim and PI3K/AKT/mTOR signaling dramatically promoted apoptosis of acute leukemia cells by downregulating eIF4B phosphorylation. Furthermore, in vitro and in vivo functional experiments showed that eIF4B played an important anti-apoptosis role in the acute leukemia cells by regulating the expression of anti-apoptotic proteins Bcl-2 and Bcl-XL. In contrast, silencing eIF4B inhibited the growth of acute leukemia cells as engrafted tumors in nude mice. Taken together, our results indicate the synergistic role of JAK/STAT5/Pim and PI3K/AKT/mTOR signaling pathways in regulating eIF4B phosphorylation in acute leukemia, and highlight eIF4B as a candidate therapeutic target for treatment of acute leukemia.