Objective:The aim of the research was to study whether microRNA-15a(miR-15a) oligonucleotide could inhibit cell growth and enhance cytarabine(Ara-C)-induced apoptosis in Raji cells.Methods:Transfecting miR-15a oligonu...Objective:The aim of the research was to study whether microRNA-15a(miR-15a) oligonucleotide could inhibit cell growth and enhance cytarabine(Ara-C)-induced apoptosis in Raji cells.Methods:Transfecting miR-15a oligonucleotide into Raji cells with LipofectamineTM 2000,and then combined with Ara-C.IC50 value and cell proliferation were detected by CCK8 assay;the expression levels of Bcl-2 mRNA and protein were evaluated by RT-PCR and indirect immuno-fluorescence.The apoptotic cells were observed by Hoechst Dyeing;AnnexinV/PI double dyeing method was used to detect the cell apoptotic rate by Flow Cytometry(FCM).Results:After Raji cells were transfected with miR-15a oligonucleotide for 48 h,Bcl2 protein expression levels obviously decreased,however,there was no difference in Bcl-2 mRNA levels,as compared with the control group and blank group(P < 0.05).CCK8 assay showed that miR-15a oligonucleotide decreased the cell growth at 24,48 and 72 h,moreover,miR-15a oligonucleotides combined with Ara-C obviously decreased the cell growth than miR-15a group,Ara-C group and scrambled oligonucleotides(SODN) + Ara-C group.Meanwhile,miR-15a oligonucleotides combined with Ara-C significantly decreased IC50 of Ara-C(10.41 μg/mL),which were obviously lower than those of Ara-C group(15.43 μg/mL) and SODN plus Ara-C group(14.92 μg/mL).Plenty of apoptotic cells could be seen with Hoechst dyeing.AnnexinV/PI double dying assays by FCM indicated that the cell apoptotic rates in earlier period and late period of miR-15a + Ara-C group were 20.93% and 25.27%,respectively,which were obviously higher than those of miR-15a group,Ara-C group and SODN plus Ara-C group.Conclusion:miR-15a oligonucleotides can inhibit cell growth and enhance Ara-C-induced apoptosis in Raji cells.展开更多
基金Supported by the grants from the Natural Science Program Foundation of the Guangdong Province (No.04010446)the Overseas Chinese Affairs Office of the State Council Key Discipline Construction Fund (No.51205002)
文摘Objective:The aim of the research was to study whether microRNA-15a(miR-15a) oligonucleotide could inhibit cell growth and enhance cytarabine(Ara-C)-induced apoptosis in Raji cells.Methods:Transfecting miR-15a oligonucleotide into Raji cells with LipofectamineTM 2000,and then combined with Ara-C.IC50 value and cell proliferation were detected by CCK8 assay;the expression levels of Bcl-2 mRNA and protein were evaluated by RT-PCR and indirect immuno-fluorescence.The apoptotic cells were observed by Hoechst Dyeing;AnnexinV/PI double dyeing method was used to detect the cell apoptotic rate by Flow Cytometry(FCM).Results:After Raji cells were transfected with miR-15a oligonucleotide for 48 h,Bcl2 protein expression levels obviously decreased,however,there was no difference in Bcl-2 mRNA levels,as compared with the control group and blank group(P < 0.05).CCK8 assay showed that miR-15a oligonucleotide decreased the cell growth at 24,48 and 72 h,moreover,miR-15a oligonucleotides combined with Ara-C obviously decreased the cell growth than miR-15a group,Ara-C group and scrambled oligonucleotides(SODN) + Ara-C group.Meanwhile,miR-15a oligonucleotides combined with Ara-C significantly decreased IC50 of Ara-C(10.41 μg/mL),which were obviously lower than those of Ara-C group(15.43 μg/mL) and SODN plus Ara-C group(14.92 μg/mL).Plenty of apoptotic cells could be seen with Hoechst dyeing.AnnexinV/PI double dying assays by FCM indicated that the cell apoptotic rates in earlier period and late period of miR-15a + Ara-C group were 20.93% and 25.27%,respectively,which were obviously higher than those of miR-15a group,Ara-C group and SODN plus Ara-C group.Conclusion:miR-15a oligonucleotides can inhibit cell growth and enhance Ara-C-induced apoptosis in Raji cells.
