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Study on the Effect of JARID1B Gene Targeting SiRNA in HL-60 Cell Line

Author: HanHuiDan
Tutor: MaXuDong
School: Fujian Medical
Course: Internal Medicine
Keywords: RNA interference JARID1B HL-60 histone methylation histone acetylation
CLC: R733.71
Type: Master's thesis
Year: 2011
Downloads: 92
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Abstract


Objective: To study JARID1B gene affecting the cell proliferation, apoptosis and histone modulation in HL-60 cells line, we design small interfering RNA（siRNA） targeting JARID1B gene.Methods:⑴Four siRNA segments targeting JARID1B gene were designed and transfected into HL-60 cells by LipofectamineTM2000. The optimal segment was screened by RT-PCR.⑵Cell growth affected by JARID1B siRNA was determined by MTT. Cell apoptosis was measured by Flow cytometry.⑶The expression of BCL-2, procaspase-9, procaspase-3, P27 and C-myc and the expression of histone methylation of H3K4 and histone acetylation of H3, H4 were detected by Western blot. Results:⑴The optimal segment is screened out from four siRNA segmentsdesigned for JARID1B. The sequence is Sense5’-GGAGCUAUUCAAUUAACUATT -3’Anti-sense5’-UAGUUAAUUGAAUAGCUCCAG-3’.⑵JARID1B siRNA upregula -tes P27 and surpresses the proliferation in HL-60 cells. The expression of Bcl-2, procaspase-9,procaspase-3,C-myc is lower and cells apoptosis is induced.⑶JARID1B siRNA upregulates histone methylated H3K4 remarkbly and histone acetylation of H3 slightly. The change of histone acetylation of H4 is not seen.Conclusions:⑴LipofectamineTM2000 transfected siRNA for JARID1B gene into HL-60 cells has high efficiency.⑵JARID1B siRNA inhibites cell growth and induces cell apoptosis in HL-60 cell line. It might be a new therapeutic target in human leukemia.⑶JARID1B siRNA upregulates histone methylated H3K4 and acetylation of H3. It doesn’t affect histone acetylation of H4. The mechanism of JARID1B effect on histone acetylation need to be further studied.

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CLC: > Medicine, health > Oncology > Hematopoietic and lymphoid neoplasms > Leukemia > Acute leukemia
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