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山萘酚对人肝癌Bel-7402细胞增殖、迁移、侵袭及凋亡的影响
Effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells
文章发布日期:2020年11月13日  来源:  作者:仲富瑞,程宦立,张浩,等  点击次数:2697次  下载次数:31次

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【摘要】:目的 探讨山萘酚对人肝癌Bel-7402细胞增殖、迁移、侵袭及凋亡的影响,并探讨其相关分子机制。 方法 将体外培养的肝癌Bel-7402细胞随机分为对照组和低、中、高浓度实验组,实验组分别用低、中、高浓度山萘酚(25、50、100 μmol/L)处理,对照组予以等量二甲基亚砜处理。用CCK-8法检测山萘酚对Bel-7402细胞活力的影响;平板克隆检测山萘酚对细胞克隆形成能力的影响;细胞划痕和Transwell小室侵袭实验检测山萘酚对细胞迁移和侵袭的影响;Western Blot检测凋亡及周期相关蛋白表达情况。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 对照组和低、中、高浓度实验组的细胞活力在处理24 h后分别为(100.00±2.72)%、(75.70±2.42)%、(62.79±2.45)%、(43.41±2.11)%,与对照组比较,各实验组细胞活力均明显下降(P值均<0.05)。各组细胞克隆形成数分别为923.3±35.2、682.7±24.4、464.0±22.0、327.3±14.0,与对照组比较,各实验组细胞克隆形成能力均明显下降(P值均<0.05)。处理24 h后,各组的相对迁移率分别为(100.00±1.11)%、(63.33±1.16)%、(51.72±3.23)%、(37.18±2.71)%,穿膜细胞数目分别为212.0±3.0、134.0±2.0、71.0±2.0、34.0±1.0,与对照组比较,各实验组相对迁移率和穿膜细胞数均明显下降(P值均<0.05)。处理48 h后,与对照组比较,低、中、高浓度实验组抗凋亡蛋白Bcl-2的表达水平均明显降低(P值均<0.05),促凋亡蛋白Bax表达水平均明显升高(P值均<0.05),周期蛋白CyclinD1表达水平均明显降低(P值均<0.05)。结论 山萘酚可抑制人肝癌Bel-7402细胞的增殖、迁移和侵袭能力,促进其凋亡,作用机制可能与调控凋亡蛋白Bax/ Bcl-2以及下调周期蛋白CyclinD1的表达有关。
【Abstract】:Objective To investigate the effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells and related molecular mechanism. Methods Hepatoma Bel-7402 cells cultured in vitro were randomly divided into control group and low-, middle-, and high-concentration experimental groups. The experimental groups were treated with low-, middle-, and high-concentration kaempferol (25, 50, and 100 μmol/L), and the control group was treated with an equal volume of dimethyl sulfoxide. CCK-8 assay was used to observe the effect of kaempferol on the viability of Bel-7402 cells; plate colony formation assay was used to evaluate the effect of kaempferol on cell colony formation ability; wound healing assay and Transwell chamber were used to observe the effect of kaempferol on cell migration and invasion; Western blot was used to measure the expression of apoptosis- and cycle-related proteins. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. Results After 24 hours of treatment, the cell viability was 100.00%±2.72% in the control group and 75.70%±2.42%, 62.79%±2.45%, and 43.41%±2.11%, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell viability (all P<0.05). The number of cell colonies was 923.3±35.2 in the control group and 682.7±24.4, 464.0±22.0, and 327.3±14.0, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell colony formation ability (all P<0.05). After 24 hours of treatment, the relative migration rate was 100.00%±1.11% in the control group and 63.33%±1.16%, 51.72%±3.23%, and 37.18%±2.71%, respectively, in the low-, middle-, and high-concentration experimental groups, and the number of transmembrane cells was 212.0±3.0 in the control group and 134.0±2.0, 71.0±2.0, and 34.0±1.0, respectively, in the low-, middle-, and high-concentration experimental groups; compared with the control group, the experimental groups had significant reductions in relative migration rate and number of transmembrane cells (all P<0.05). After 48 hours of treatment, compared with the control group, the low-, middle-, and high-concentration experimental groups had a significant reduction in the expression of the anti-apoptotic protein Bcl-2 (all P<0.05), a significant increase in the expression of the pro-apoptotic protein Bax (all P<0.05), and a significant reduction in the expression of C<italic/>yclinD1 (all P<0.05). Conclusion Kaempferol can inhibit the proliferation, migration, and invasion of human hepatoma Bel-7402 cells and promote the apoptosis of such cells, possibly by regulating the apoptosis proteins Bax and Bcl-2 and downregulating the expression of CyclinD1.
【关键字】:山萘酚; 肝肿瘤; 细胞增殖; 细胞运动; 细胞凋亡
【Key words】:kaempferol; liver neoplasms; cell proliferation; cell movement; apoptosis
【引证本文】:ZHONG FR, CHENG HL, ZHANG H, et al. Effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells[J]. J Clin Hepatol, 2020, 36(12): 2725-2729. (in Chinese)
仲富瑞, 程宦立, 张浩, 等. 山萘酚对人肝癌Bel-7402细胞增殖、迁移、侵袭及凋亡的影响[J]. 临床肝胆病杂志, 2020, 36(12): 2725-2729.

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