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目的探讨敲低膜联蛋白A7表达对人肝癌细胞系Hep G2细胞凋亡及凋亡相关蛋白Bcl-2和Bax表达的影响。方法将Hep G2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RT-PCR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Hep G2细胞的凋亡,降低Bcl-2和Bax的比值。
Objective To investigate the effect of knockdown of annexin A7 on the apoptosis and the expression of Bcl-2 and Bax in human hepatocellular carcinoma cell line Hep G2. Methods Hep G2 cells were seeded on a 6-well plate and divided into three groups: siRNA interference group, negative control group and blank control group, in which the interfering group transfected siRNA targeting annexin A7 only, negative control group transfected only negative Control siRNA, blank control group only transfection reagent. Annexin A7 was identified by Western blotting at 48h after transfection can be inhibited to a maximum extent, so 48h after transfection using flow cytometry to detect the apoptosis rate of each group, by immunohistochemistry, Western blotting and RT-PCR Detection of Bcl-2 and Bax protein and mRNA expression. Results Compared with the negative control group and the blank control group, the apoptosis rate of siRNA interference group was significantly increased (P <0.05), the expression of Bcl-2 protein and mRNA were significantly decreased (P <0.05), while Bax protein and mRNA were No significant change (P> 0.05). Conclusion Knockdown of annexin A7 can promote the apoptosis of Hep G2 cells and decrease the ratio of Bcl-2 to Bax.