转染APOBEC3A基因对宫颈癌Hela细胞生物学行为的影响

Effects of Human APOBEC3A on Biological Behaviors of Cervical Cancer Hela Cells

【目的】探讨载脂蛋白B mRNA编辑酶催化多肽样蛋白3A(APOBEC3A)表达对宫颈癌Hela细胞体外增殖、凋亡及迁移能力的影响。【方法】根据Hela细胞是否转染APOBEC3A真核表达质粒,分A3A特异性转染组(A3A组),空白质粒转染组(阴性对照组,M45)和未转染组(空白组,Hela)。利用Western blotting检测A3A特异性转染组A3A蛋白的表达;而后使用CCK8法、流式细胞仪分析法和异质粘附、Transwell法分别检测A3A对Hela细胞增殖、凋亡、粘附及迁移等生物学行为的影响。【结果】A3A组与阴性对照组比较,Hela细胞的增殖、粘附、迁移能力下降,凋亡率增加,差异均有统计学意义(P<0.05);阴性对照组与空白组比较,增殖能力、凋亡率、粘附及迁移能力无明显差异(P>0.05)。【结论】A3A基因可能具有抑制Hela细胞增殖、粘附、迁移并诱导凋亡的功能,为研究A3A基因在宫颈癌中的作用及其生物治疗提供了理论基础。

[Objective] To investigate the effect of human apolipoprotein B mRNA-editing catalytic polypeptide-like protein 3A on the biological behaviors （such as growth,proliferation,apoptosis,migration） of cervical cancer Hela cells.[Methods] The plasmid containing APOBEC3A gene was transfected into Hela cells （A3A）.Blank plasmid-transfected Hela cells （M45/neo） and untransfected Hela cells were used as controls.The expression of A3A protein was detected by Western blotting.The impact of A3A on the biological behaviors of cervical cancer Hela cells,including cell proliferation,cell cycle,apoptosis,and migration,was subsequently analyzed using CCK8 assay,flow cytometry,and a Transwell chamber migration assay.[Result] After transfection,the protein of A3A was expressed successfully in the A3A-transfected group by the detection of Western blotting.The cell viability,proliferation capacity,and migration ability of Hela cells in the A3A-transfected group were significantly lower than those of Hela cells in the negative control group and the blank control group （P ＜ 0.05）.The percentage of apoptotic Hela cells in the A3A-transfected group was significantly higher than those of Hela cells in the negative control group and the blank control group （P ＜ 0.05）.There was no significant difference in the viability,proliferation capacity,apoptosis,and migration ability of Hela cells between the negative control group and the blank control group （P ＞ 0.05）.[Conclusion] Transfection of A3A can inhibit the proliferation and the growth of Hela cells,thereby suppressing the migration ability of Hela cells and promoting their apoptosis.This study can provide an experimental basis for future biological therapies for human cervical cancer by A3A.