| Objective: To determine the expression level of long chain non-coding RNA PAN3-AS1（lnc RNA PAN3-AS1）in patients with AML,and to analyze its effects on the clinical features,therapeutic efficacy and prognosis of AML patients.Furthermore,to observe the effect of PAN3-AS1 on the biological function of AML cell lines,and to explore the correlation between PAN3-AS1 and PAN3.Methods:1.Bone marrow specimens of 45 patients with AML and 25 patients with iron deficiency anemia enrolled in the Department of Hematology of first people’s Hospital of Jining City from February 2020 to March 2022 were collected.Mononuclear cells were isolated and total RNA was extracted.The expression of PAN3-AS1 was detected by qRT-PCR,and the difference of PAN3-AS1 expression between the two groups was compared.According to the median value of PAN3-AS1 expression,AML patients were divided into high expression group and low expression group.The differences of clinical features,therapeutic efficacy and prognosis between the two groups were compared.2.HL-60 cell line was cultured in vitro.Si RNA interference technique knock down the expression of PAN3-AS1,and lentivirus transfection technique was used to overexpress PAN3-AS1.Then CCK8 proliferation test,transwell test and flow cytometry were used to detect the effects of PAN3-AS1 on the proliferation,migration,invasion,apoptosis and cell cycle biological function of HL-60 cell line.3.qRT-PCR method was used to detect the expression of PAN3-AS1 and PAN3 in bone marrow mononuclear cells of patients with AML,and the correlation between the two expression levels was analyzed.To further detect the expression level of PAN3 in AML cells after PAN3-AS1 overexpression.Results:1.The expression of PAN3-AS1 in patients with AML and its effect on clinical features1.1Compared with the control group,the expression of PAN3-AS1 was significantly increased in newly diagnosed AML patients（P=0.011）.1.2 Compared with the low expression group,the patients with high expression of PAN3-AS1 had a higher proportion of bone marrow blasts（P=0.040）and were more likely to express CD4,CD9 and CD36（P=0.027、0.031、0.027）,and low expression of CD7（P=0.009）.And the high expression group were more likely to have FLT3-ITD mutation（P=0.017）.1.3 There was no significant difference in the efficacy and prognosis of induction therapy between the low expression group and the high expression group of PAN3-AS1（P>0.05）.2.Effect of PAN3-AS1 on biological characteristics of AML cell line2.1 CCK8 assay of cell proliferation showed that compared with the control group,the proliferation ability of HL-60 cells in the PAN3-AS1 interference group decreased（P<0.05）,while the cell proliferation ability in the PAN3-AS1 overexpression group was stronger than that in the control group（P<0.05）.2.2Transwell migration and invasion assay showed that the migration and invasion ability of HL-60 cells in PAN3-AS1 interference group was lower than that in control group（P<0.05）,while the migration and invasion ability in PAN3-AS1 overexpression group was stronger than that in control group（P<0.05）.2.3 Flow cytometry showed that the proportion of apoptosis in PAN3-AS1 overexpression group was significantly lower than that in control group,and the cells were blocked in S phase（P<0.05）.3.Correlation between PAN3-AS1 and PAN3 expressionThere was a positive correlation between the expression of PAN3-AS1 and PAN3 in bone marrow samples of patients with AML（r=0.704,P<0.001）,and the expression of PAN3 in AML cells increased after overexpression of PAN3-AS1（P<0.05）.Conclusion:1.PAN3-AS1 was highly expressed in AML patients,and the high expression group had higher proportion of bone marrow primordial cells,higher expression of CD4,CD9,CD36 and low expression of CD7.And the high expression group were more likely to have FLT3-ITD mutation2.PAN3-AS1 is highly expressed in AML cell line,and can promote cell proliferation,migration,invasion,inhibit apoptosis,relieve cell cycle arrest and play the role of oncogenes.3.PAN3-AS1 may participate in the occurrence and development of AML by regulating the expression of PAN3. |
