Liu Yang, Zhou Xiaofang, Wen Jinsong, Deng Shishan, Liu Hai
Department of Otolaryngology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
Department of Otolaryngology, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China. Email:
Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2014 Apr;49(4):326-9.
To study the effect of annexin A5 on the apoptosis of laryngeal cancer cells.
Special siRNAs were used to knock annexinA5 down in Hep-2 cell, and RT-PCR and Western blot were applied to identify the efficacy of RNA interference. The flow cytometry assay was performed to detect the Hep-2 cell apoptosis.
RT-PCR analysis showed that the relative mRNA expression of annexin A5 in siRNA group, negative control group, Lipofectamine 2000 group and blank control group were 0.70 ± 0.03, 1.18 ± 0.05, 1.17 ± 0.06 and 1.23 ± 0.07. The relative mRNA expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t = -14.77, t = -13.23, t = -12.99, P < 0.05).In Western blot assay, the trend of protein expression level was consistent with the mRNA expression levels of annexin A5. The relative levels of proteins in siRNA group, negative control group, Lipofectamine 2000 group and blank control group were shown 1.21 ± 0.03, 3.88 ± 0.06, 3.87 ± 0.02 and 3.95 ± 0.08. The relative protein expression of annexin A5 in siRNA group was significantly decreased than contrast groups(t = -70.34, t = -150.62, t = -56.32, P < 0.05). At the same time in flow cytometry the apoptotic rate of siRNA group, negative control group, Lipofectamine 2000 group and blank control group were 4.43% ± 0.12%, 13.67% ± 0.22%, 13.66% ± 0.12% and 13.35% ± 0.13%, the difference between the siRNA group and contrast groups was statistically significant(t = -62.50, t = -14.16, t = -11.47, P < 0.05).So after RNA interference, expression of annexin A5 decreased, and the results in the apoptosis inhibition of Hep-2 cell.
Annexin A5 promotes apoptosis of Hep-2 cells, and it may be a potential therapeutic target for the laryngeal cancer.
研究膜联蛋白A5对喉癌细胞凋亡的影响。
使用特异性小干扰RNA（siRNAs）敲低Hep-2细胞中的膜联蛋白A5，应用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法（Western blot）鉴定RNA干扰效果。采用流式细胞术检测Hep-2细胞凋亡情况。
RT-PCR分析显示，siRNA组、阴性对照组、脂质体2000组和空白对照组中膜联蛋白A5的相对mRNA表达量分别为0.70±0.03、1.18±0.05、1.17±0.06和1.23±0.07。siRNA组中膜联蛋白A5的相对mRNA表达量较对照组显著降低（t=-14.77，t=-13.23，t=-12.99，P<0.05）。蛋白质免疫印迹分析中，膜联蛋白A5的蛋白质表达水平趋势与mRNA表达水平一致。siRNA组、阴性对照组、脂质体2000组和空白对照组中蛋白质的相对水平分别为1.21±0.03、3.88±0.06、3.87±0.02和3.95±0.08。siRNA组中膜联蛋白A5的相对蛋白质表达量较对照组显著降低（t=-70.34，t=-150.62，t=-56.32，P<0.05）。同时，流式细胞术检测显示，siRNA组、阴性对照组、脂质体2000组和空白对照组的凋亡率分别为4.43%±0.12%、13.67%±0.22%、13.66%±0.12%和13.35%±0.13%，siRNA组与对照组之间的差异具有统计学意义（t=-62.50，t=-14.16，t=-11.47，P<0.05）。因此，RNA干扰后，膜联蛋白A5表达降低，结果导致Hep-2细胞凋亡受到抑制。
膜联蛋白A5促进Hep-2细胞凋亡，可能是喉癌潜在的治疗靶点。
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