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作 者:冯晶晶[1] 雷炜[1] 姚如永[2] 赵园园[1] 阎超[1]
机构地区:[1]青岛大学医学院附属医院肿瘤科,山东青岛266003 [2]青岛大学医学院附属医院中心实验室,山东青岛266003
出 处:《现代生物医学进展》2012年第18期3446-3449,共4页Progress in Modern Biomedicine
摘 要:目的:研究靶向survivin的(小分子干扰RNA)siRNA和(氟尿嘧啶)5-FU联用对肝癌细胞HepG2的增殖抑制及凋亡的影响。方法:将HepG2细胞分为空白对照组、阴性对照组、5-FU处理组、siRNA转染组、5-FU+siRNA转染组。转染采用脂质体法。RT-PCR法检测HepG2细胞survivinmRNA转录水平;MTT法检测靶向survivin的siRNA和5.FU对HepG2细胞增殖的抑制作用;流式细胞术检测HepG2细胞凋亡情况。结果:空白对照组、阴性对照组、5-FU处理组survivinmRNA表达无明显变化(P〉0.05),siRNA转染组、5-FU+siRNA转染组survivinmRNA表达明显下降(F=280.326,q=4.72-7.34,P〈0.05)。5-FU+siRNA转染组增殖抑制率为51.58%±1-35%,与其它各组相比抑制率明显增高(F=280.326,q=5.27-9.84,P〈0.05)。5-Fu+siRNA组与其它各组相比细胞凋亡率明显增高(F=13568.68,q=110.47-327.16,P〈0.01)。结论:将靶向survivin的siRNA和5一Fu联合应用可以显著抑制肝癌细胞survivin基因表达,并协同抑制HepG2细胞增殖,共同发挥诱导细胞凋亡作用。Objective: To investigate the anti-proliferation effect of (small interference RNA) siRNA against survivin combined with(fluorouracil) 5-FU on human liver cancer cell line HepG2. Methods: HepG2 cells cultures were divided into five groups: blank con- trol group, negative control group, 5-FUgroup, siRNA-survivin group, siRNA-survivin+5-FU group. Lipofectamine TM 2000 was used to transfect HepG2 cell. The expression of survivin mRNA was detected by RT-PCR. Inhibition rate of each group on HepG2 was assayed by MTT method, and apoptosis was analyzed by flow cytometry respectively. Results: The experiment of RT-PCR showed that the ex- pression of survivin mRNA in siRNA-survivin group and siRNA-survivin+5-FU group were lower than that in other groups(P〈0.05), but its levels in blank control group, negative control group and 5-FU group had no significant change (P〉0.05). The MTT assay indicated that the antiproliferation rate of siRNA-survivin+5-FU group was 51.38%+ 1.35. The antiproliferation rate of this group was higher than that in any other groups(P〈0.05). Flow cytometry results indicated that the apoptosis rate of siRNA-survivin+5-FU group was also higher than any other groups (P〈0.05). Conclusion: siRNA against survivin combined with 5-FU can specifically suppress its expression, inhibit the proliferation of HepG2 and induce apoptosis in common.
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