The invention also discloses application of serine integration factor 2 in MLL fusion gene leukemia
A serine and leukemia technology, applied in the field of biomedicine, can solve the problems of easy recurrence, complicated pathogenesis, poor prognosis, etc., and achieve the effect of prolonging the survival period and inhibiting cell proliferation
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Embodiment 1
[0036] Example 1 Serine Integration Factor 2 (SERINC2) Expression Analysis
[0037] The present invention discovers a gene SERINC2 that is specifically highly expressed in MLL-R leukemia and lowly expressed in normal physiological conditions and other types of leukemia. This gene encodes five different transmembrane proteins ( figure 1 A). Among them, transcript 1 is an 11 transmembrane protein ( figure 1 B). Its nucleotide sequence is shown in SEQ ID NO:1; its amino acid sequence is shown in SEQ ID NO:2.
[0038] In order to further confirm the expression specificity of SERINC2 in MLL-R leukemia, we collected a batch of bone marrow samples from the First Affiliated Hospital of Sun Yat-sen University for detection and analysis, including 51 cases of MLL wild-type (MLL-wt) leukemia samples, And 26 cases of MLL-R leukemia samples. All sample collection was approved by the Ethics Committee of Sun Yat-Sen University and informed consent was obtained from the patients. SERINC...
Embodiment 2
[0042] Example 2 Functional identification of SERINC2 in MLL-R leukemia
[0043] In order to solve the core problem of SERINC2 participating in the regulation of MLL-R leukemia, we intend to further study the function of SERINC2 on MLL-R leukemia. Therefore, we selected MLL-R leukemia cell lines, RS4-11 (MLL-AF4) and MV4-11 (MLL-AF4) as the research objects. Two different siRNA sequences were designed for SERINC2 by siRNA interference technology.
[0044] Forward sequence of siRNA-1: 5'-GGTCAGCCTATCCAGTAT dTdT-3' (SEQ ID NO: 5);
[0045] Reverse sequence of siRNA-1: 5'-CCAGTCGGGATAGGTCATA dTdT-3' (SEQ ID NO: 6);
[0046] Forward sequence of siRNA-2: 5'-TGACGCTCACCAACTGGTA dTdT-3' (SEQ ID NO: 7);
[0047] Reverse sequence of siRNA-2: 3'-ACTGCGAGTGGTTGACCAT dTdT-5' (SEQ ID NO: 8).
[0048] In the above two MLL-R leukemia cell lines, SERINC2 was knocked out respectively, and the effect of SERINC2 knockdown and expression in MLL-R leukemia was detected by qRT-PCR, and the cell...
Embodiment 3
[0059] Example 3 SERINC2 regulates the mTOR pathway
[0060] SERINC2 is a protein of the serine integrator family, which was identified and named in 2005 and proved to be related to serine integration in lipid synthesis. The present invention finds that SERINC2 is localized on autophagy lysosomes ( Figure 5A). SERINC2 is a member of the amino acid transporter family, therefore, we speculate that on the autophagosome membrane, SERINC2 mediates the activation of the mTOR pathway by sensing amino acid changes. To verify our speculation, we knocked down SERINC2 using shRNA in RS4-11 and MV4-11, and detected the phosphorylation level of S6K protein directly downstream of mTOR. We used amino acid starvation re-induction experiments, in which 1640 medium without amino acids was used for starvation for 50 minutes, and medium containing all amino acids was added for induction for 30 minutes. First of all, this study first found that SERINC2 is localized on autolysosomes and its loc...
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