2013•白血病·淋巴瘤Requires access

Effects of antisense oligonucleotides targeting microRNA-17 on human leukemic K562 cells

Jun-chun Zhou, Jiahua Liu, Jian‐Mei Li

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Abstract

Objective To explore the inhibitory effect of anti-miRNA-17 oligonucleotide on leukemic K562 cells.Methods K562 cells were transfected with anti-miRNA-17 oligonucleotide,cell viability was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliunbromide (MTT) assay.Apoptosis was detected by flow cytometry,expression of miRNA-17 in K562 cells was measured by real-time PCR.Results MTT results showed transfection of antisense nucleic acid significantly decreased cell proliferation activity,after 24,48,72 h they were respectively 0.8719±0.001,0.7102±0.002,0.5507±0.001,the difference being statistically significant (P < 0.05) when compared to the random control (t =182.575,269.77,660.4) or control group (t =537.98,571.20,1230.51).FCM test results showed that after 48 h of transfecting antisense nucleic acid apoptosis rate was (20.14 ± 0.01) %,and was statistically significant compared to the randomized control or blank control group (t =2347.6,2568.2,P < 0.01).Fluorescence real-time quantitative PCR confirmed antisense nucleic acid significantly decreased the relative expression level of miR-17 in K562 cells (0.07).The difference was statistically significant compared to the random group or blank group (1,1.01) (t =148.63,147.04,P < 0.05).Conclusion Targeted inhibition of miR-17 with oligonucleotide can suppress K562 cell growth and induce apoptosis. Key words: microRNA-17; Antisense oligonucleotides; K562 cells; Apoptosis

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Objective To explore the inhibitory effect of anti-miRNA-17 oligonucleotide on leukemic K562 cells.Methods K562 cells were transfected with anti-miRNA-17 oligonucleotide,cell viability was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliunbromide (MTT) assay.Apoptosis was detected by flow cytometry,expression of miRNA-17 in K562 cells was measured by real-time PCR.Results MTT results showed transfection of antisense nucleic acid significantly decreased cell proliferation activity,after 24,48,72 h they were respectively 0.8719±0.001,0.7102±0.002,0.5507±0.001,the difference being statistically significant (P < 0.05) when compared to the random control (t =182.575,269.77,660.4) or control group (t =537.98,571.20,1230.51).FCM test results showed that after 48 h of transfecting antisense nucleic acid apoptosis rate was (20.14 ± 0.01) %,and was statistically significant compared to the randomized control or blank control group (t =2347.6,2568.2,P < 0.01).Fluorescence real-time quantitative PCR confirmed antisense nucleic acid significantly decreased the relative expression level of miR-17 in K562 cells (0.07).The difference was statistically significant compared to the random group or blank group (1,1.01) (t =148.63,147.04,P < 0.05).Conclusion Targeted inhibition of miR-17 with oligonucleotide can suppress K562 cell growth and induce apoptosis. Key words: microRNA-17; Antisense oligonucleotides; K562 cells; Apoptosis

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Available abstract

Objective To explore the inhibitory effect of anti-miRNA-17 oligonucleotide on leukemic K562 cells.Methods K562 cells were transfected with anti-miRNA-17 oligonucleotide,cell viability was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra-zoliunbromide (MTT) assay.Apoptosis was detected by flow cytometry,expression of miRNA-17 in K562 cells was measured by real-time PCR.Results MTT results showed transfection of antisense nucleic acid significantly decreased cell proliferation activity,after 24,48,72 h they were respectively 0.8719±0.001,0.7102±0.002,0.5507±0.001,the difference being statistically significant (P < 0.05) when compared to the random control (t =182.575,269.77,660.4) or control group (t =537.98,571.20,1230.51).FCM test results showed that after 48 h of transfecting antisense nucleic acid apoptosis rate was (20.14 ± 0.01) %,and was statistically significant compared to the randomized control or blank control group (t =2347.6,2568.2,P < 0.01).Fluorescence real-time quantitative PCR confirmed antisense nucleic acid significantly decreased the relative expression level of miR-17 in K562 cells (0.07).The difference was statistically significant compared to the random group or blank group (1,1.01) (t =148.63,147.04,P < 0.05).Conclusion Targeted inhibition of miR-17 with oligonucleotide can suppress K562 cell growth and induce apoptosis. Key words: microRNA-17; Antisense oligonucleotides; K562 cells; Apoptosis

Key concepts: K562 cells, Apoptosis, Molecular biology, Transfection, Oligonucleotide, Flow cytometry, Nucleic acid, microRNA

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