To Investigate The Effects Of HOXA5 Gene On The Proliferation Of AML Cells By The CRISPR-Cas9 Gene Editing Technology

Objectives: The Homeobox A5(HOXA5)gene acts as an anti-oncogene in gastric cancer,hepatocellular carcinoma,cervical cancer,and ovarian cancer,and as an oncogene in esophageal squamous cell carcinoma and glioblastoma.The specific mechanism of HOXA5 gene in tumors remains controversial.The relevant researches on the role of HOXA5 gene in acute myeloid leukemia(AML)is few.The project based on the expression of HOXA5 gene in acute myeloid leukemia intends to construct acute myeloid leukemia cell lines with stable HOXA5 gene knockout through CRSIPR-Cas9 gene editing technology,and to determine the effect of HOXA5 gene knockout on the proliferation of acute myeloid leukemia cells.These studies preliminarily explore the role of HOXA5 gene in the pathogenesis of acute myeloid leukemia.Methods: 1.Clinic trial: Quantitative real-time PCR(q RT-PCR)and Western blot were utilized to detect the expression of HOXA5 in bone marrow mononuclear cells of hematological system non-tumor patients and acute myeloid leukemia patients.2.Cell experiment: A stable acute myeloid leukemia cell line with HOXA5 gene knockout was constructed via CRISPR-Cas9 gene editing technology.The transfected cells were screened via the puromycin screening experiment.Finally,the expression of HOXA5 m RNA and protein levels were detected by q RT-PCR and Western blot,and KO-HOXA5-THP-1 cells were obtained.CCK-8 assay was utilized to explore the proliferation of THP-1 cells and KO-HOXA5-THP-1 cells.Results: 1.The high expression of HOXA5 was reflected in m RNA and protein levels in bone marrow mononuclear cells of acute myeloid leukemia patients compared with non-tumor hematological patients.(P <0.05).2.The acute myeloid leukemia cell line with stable knockout of HOXA5 gene(KO-HOXA5-THP-1)can be obtained via CRISPR-Cas9 gene editing technology.Theproliferation ability of the cell line is distinctly inhibited compared with THP-1 cells(P<0.05).Conclusions: The high expression of m RNA and protein levels of HOXA5 gene is in acute myeloid leukemia cells.Knockout of HOXA5 gene by CRISPR-Cas9 gene editing technology significantly affects the proliferation of acute myeloid leukemia cells.The study provides a new potential therapeutic target for the precision treatment of acute myeloid leukemia.