INVESTIGATION OF APOPTOSIS INDUCTION USING A SUITABLE SIRNA EXPRESSION VECTOR IN CULTURED K562 CELL LINE

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JAVADI HAMID REZA; ZAREI MAHMOUDABADI A.; KAMALI MAHDI; HOJATI ZAHRA; NAJAFI ALI

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Journal: KOWSAR MEDICAL JOURNAL Year:2008 | Volume:13 | Issue:3 Page(s): 185-196

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Title

INVESTIGATION OF APOPTOSIS INDUCTION USING A SUITABLE SIRNA EXPRESSION VECTOR IN CULTURED K562 CELL LINE

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Keywords

SIRNAQ3

CMLQ3

PRNA.H1-1/NEOQ3

BCR/ABLQ3

APOPTOSISQ3

Abstract

Objective: Design of a suitable SIRNA against BCR/ABL and using an expressing vector (pRNA-H1.1-Neo) for expression of SIRNA in transfected K562 cell line and evaluate of APOPTOSIS induction value.Matherials and methods: select an suitable sequence from chimeric BCR/ABL mRNA and cloned in expression vector (pRNAH1.1/Neo) as a ShRNA expressing DNA. Then evaluate the cloning by using PCR and sequencing of insert. Cloned vector was transfected to K562 by lipofectamin 2000 protocol and a miss match SIRNA and uncloned vector used as ngative control. The efficacy of transfected method was investigated using GFP expressing vecapoptosis was investigated in transfected cells using cell death detection ELISA kit.Results: The PCR results, electrophoresis and sequencing shown the accuracy of cloned vector and GFP expression with green fleurence confirmed the transfection method with efficacy equal 44%. The APOPTOSIS value in transfected cells with cloned vector in compared with control cells was significantly increased in 24,48 and 72h after transfection. The APOPTOSIS induction was time dependent and 72h after transfection it was 36±3.2% (p<0.0) but in normal ,control vector and negative control cells was respectively, %5.05±0.23, %5.2±0.86 and %4.8±0.6 which was constant with time. Wherase, in cells which transfected with cloned vector, the induction of APOPTOSIS rate was timely and comparable with reduction in BCR/ABL mRNA.Conclusion: The expression pRNA. H1-1/Neo vector was suitably coloned with BCR/ABL ShRNA andintracellular SIRNA was expressed which reduced BCR/ABL mRNA and therefore induced APOPTOSIS in K562 cell line. Therefore, specific SIRNA could be attention as a new molecular trapeiutic method for CML.

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APA: Copy

JAVADI, HAMID REZA, ZAREI MAHMOUDABADI, A., KAMALI, MAHDI, HOJATI, ZAHRA, & NAJAFI, ALI. (2008). INVESTIGATION OF APOPTOSIS INDUCTION USING A SUITABLE SIRNA EXPRESSION VECTOR IN CULTURED K562 CELL LINE. KOWSAR MEDICAL JOURNAL, 13(3), 185-196. SID. https://sid.ir/paper/32764/en

Vancouver: Copy

JAVADI HAMID REZA, ZAREI MAHMOUDABADI A., KAMALI MAHDI, HOJATI ZAHRA, NAJAFI ALI. INVESTIGATION OF APOPTOSIS INDUCTION USING A SUITABLE SIRNA EXPRESSION VECTOR IN CULTURED K562 CELL LINE. KOWSAR MEDICAL JOURNAL[Internet]. 2008;13(3):185-196. Available from: https://sid.ir/paper/32764/en

IEEE: Copy

HAMID REZA JAVADI, A. ZAREI MAHMOUDABADI, MAHDI KAMALI, ZAHRA HOJATI, and ALI NAJAFI, “INVESTIGATION OF APOPTOSIS INDUCTION USING A SUITABLE SIRNA EXPRESSION VECTOR IN CULTURED K562 CELL LINE,” KOWSAR MEDICAL JOURNAL, vol. 13, no. 3, pp. 185–196, 2008, [Online]. Available: https://sid.ir/paper/32764/en

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