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敲低膜联蛋白A7对人肝癌HepG2细胞Bcl-2和Bax表达的影响
Influence of annexin A7 knockdown on expression of Bcl-2 and Bax of HepG2 cells
目的 探讨敲低膜联蛋白A7表达对人肝癌细胞系HepG2细胞凋亡及凋亡相关蛋白Bcl-2和Bax表达的影响。方法 将HepG2细胞接种于6孔板，分为3组：siRNA干扰组、阴性对照组和空白对照组，其中干扰组只转染靶向膜联蛋白A7的siRNA，阴性对照组只转染阴性对照siRNA，空白对照组只加转染试剂。通过Western blotting鉴定膜联蛋白A7在转染后48h可被最大程度的抑制，于是在转染后48h采用流式细胞术检测各组细胞凋亡率，通过免疫组织化学、Western blotting和RTPCR检测Bcl-2和Bax蛋白及mRNA的表达。 结果 与阴性对照组和空白对照组相比，siRNA干扰组细胞凋亡率显著增高（P<0.05），Bcl-2蛋白和mRNA的表达均显著降低（P<0.05），而Bax蛋白和mRNA均未发生显著变化（P>0.05）。结论 敲低膜联蛋白A7可促进HepG2细胞的凋亡，降低Bcl-2和Bax的比值。
Objective To investigate the influence of annexin A7 knockdown on apoptosis and expression of Bcl-2 and Bax of HepG2 cells. Methods HepG2 cells cultured in six-well plates were divided into 3 groups, siRNA interference group, negative control group and blank control group. The siRNA interference group was transfected with the siRNA which target annexin A7, the negative control group was transfected with negative siRNA, in the blank control group only transfection reagent was added. Western blotting was used to identify the siRNA targeted to annexin A7 can highly surpress annexin A7 expression in 48h after transfection. Flow cytometry was used to detect the apoptotic rate. Immunohistochemistry and Western blotting were applied to observe the expression of Bcl-2 and Bax protein; RT-PCR was performed to detect the expression of Bcl-2 and Bax mRNA. Results The apoptotic rate increased and the expression of Bcl-2 decreased significantly compared with the negative control group and the blank control group(P<0.05), while the expression of Bax protein and mRNA had no significant difference when compared with the negative control group and the blank control group(P>0.05). Conclusion Annexin A7 knockdown can promote apoptosis of HepG2 cells, reduce the ratio of Bcl-2 and Bax.
膜联蛋白A7 / HepG2细胞 / Bcl-2 / Bax / 流式细胞术 / 人
Annexin A7 / HepG2 cell / Bcl-2 / Bax / Flow cytometry / Human
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