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目的:研究根皮素对体外培养人宫颈癌He La细胞增殖、凋亡及侵袭的影响,并探讨其可能的作用机制。方法:采用MTT法检测不同浓度及不同时间根皮素对He La细胞增殖的抑制作用,采用流式细胞仪分析根皮素对He La细胞凋亡的影响,应用Millicell小室检测根皮素处理后细胞侵袭能力的变化,采用Western blot方法检测He La细胞Survivin及caspase-3蛋白表达水平,应用RT-PCR方法检测MMP-2 mRNA及MMP-9 mRNA的表达。结果:根皮素能抑制He La细胞增殖,在一定浓度范围内呈时间和剂量依赖性(P<0.05);不同浓度根皮素处理He La细胞后能诱导其凋亡,与对照组比较差异均有统计学意义(P<0.05);Millicell小室侵袭实验结果显示:根皮素处理后细胞的穿膜数明显减少(P<0.05);随着根皮素浓度的增加,Survivin蛋白表达水平逐渐下降(P<0.05),caspase-3蛋白的表达水平逐渐增加(P<0.05);MMP-2 mRNA及MMP-9 mRNA的表达随着根皮素浓度的增加逐渐下降(P<0.05)。结论:根皮素可显著抑制人宫颈癌He La细胞增殖,可能通过抑制Survivin蛋白表达并激活caspase-3而诱导凋亡,通过下调MMP-2及MMP-9的表达而降低He La细胞的侵袭能力。
Objective: To investigate the effects of phloretin on proliferation, apoptosis and invasion of Hela cells in vitro and to explore its possible mechanism. Methods: MTT assay was used to detect the inhibitory effect of phloretin on the proliferation of HeLa cells at different concentrations and different times. The effects of phloretin on the apoptosis of HeLa cells were analyzed by flow cytometry. The cell invasion ability was detected by Western blot. The expression of Survivin and caspase-3 protein in HeLa cells was detected by Western blot. The expressions of MMP-2 mRNA and MMP-9 mRNA were detected by RT-PCR. Results: Phloretin could inhibit the proliferation of HeLa cells in a time and dose-dependent manner (P <0.05). The apoptosis of HeLa cells was induced by different concentrations of phloretin, compared with the control group (P <0.05). The results of Millicell chamber invasion assay showed that the number of transmembrane cells in the cells treated with phloretin decreased significantly (P <0.05). Survivin protein expression gradually increased with the increase of the concentration of phloretin (P <0.05). The expression of caspase-3 protein increased gradually (P <0.05). The expression of MMP-2 mRNA and MMP-9 mRNA decreased gradually with the increase of phloretin concentration (P <0.05). CONCLUSION: Phloretin can significantly inhibit the proliferation of human cervical cancer HeLa cells, which may induce apoptosis by inhibiting the expression of Survivin and activating caspase-3, and reduce the invasion of HeLa cells by down-regulating the expressions of MMP-2 and MMP-9 ability.