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Received:April 03, 2020 Published Online:January 20, 2021
Received:April 03, 2020 Published Online:January 20, 2021
中文摘要: 目的 探讨蜂毒肽对人口腔鳞状细胞癌(OSCC)细胞系Scc-15、Tca8113、Cal-27增殖性、迁移性的影响及其作用机制。
方法 以用四种不同浓度梯度(2、4、8、16 μg/ml)的蜂毒肽分别处理的OSCC细胞系为蜂毒肽组,以不含药的培养液处理的细胞系为对照组。采用MTT实验检测蜂毒肽对OSCC细胞系Scc-15、Tca8113、Cal-27增殖的抑制作用;划痕实验观察蜂毒肽对Scc-15细胞迁移能力的抑制效果;Transwell实验验证蜂毒肽对Scc-15细胞侵袭能力的抑制作用;Western blot检测蜂毒肽作用于Scc-15细胞后核因子(NF)κB、半胱氨酸天冬氨酸蛋白酶(Caspase)-3、Caspase-9的表达情况。
结果 MTT检测结果示,相对于对照组,2、4、8、16 μg/ml蜂毒肽组的Scc-15、Tca8113、Cal-27细胞增殖存活率均明显降低,且呈浓度依赖性(P均<0.01)。划痕实验结果示,在细胞划痕培养6、12和24 h时相对于对照组,8、16 μg/ml蜂毒肽组的Scc-15细胞划痕愈合率均明显降低,且呈浓度依赖性(P均<0.01)。Transwell侵袭实验结果示,对照组与16 μg/ml蜂毒肽组穿过滤过膜的Scc-15细胞数分别为(46.0±2.3)个/视野和(21.0±1.7)个/视野,蜂毒肽组显著少于对照组(P<0.01)。Western blot结果示,相对于对照组,加入蜂毒肽后,Scc-15细胞的NF-κB蛋白表达量显著下降(P<0.01)、Caspase-3和Caspase-9蛋白表达量显著升高(P均<0.01)。
结论 蜂毒肽可显著抑制OSCC细胞的增殖、迁移及侵袭能力,还可促进OSCC细胞凋亡,其作用机制可能与NF-κB的生成减少相关。
Abstract:Objective To investigate the effects of Melittin (MEL) on the proliferation and migration of human oral squamous cell carcinoma (OSCC) cell lines Scc-15,Tca8113,and Cal-27 and its mechanism.
Methods OSCC cell lines respectively treated with four different concentrations of MEL (2,4,8,16 μg/ml)were served as MEL group.Cells without treatment of MEL were served as control group.MTT assay was used to measure the inhibitory effect of MEL on the proliferation of OSCC cell lines Scc-15,Tca8113 and Cal-27.
Scratch test and Transwell test were used to detect the inhibitory effects of MEL on migration and invasion of Scc-15 cells respectively.
Western blot was used to detect the expressions of nuclear factor (NF) κB,Caspase-3 and Caspase-9 in Scc-15 cells treated with MEL.
Results MTT assay results showed that compared with control group,the proliferation and survival rates of Scc-15,Tca8113 and Cal-27 cells in 2-,4-,8-,16- μg/ml MEL groups significantly decreased in a concentration-dependent manner (all P<0.01).Scratch test showed that scratch healing rates of Scc-15 cells in 8 and 16 μg/ml MEL groups were significantly lower than that in control group in a concentration-dependent manner at 6-,12- and 24-h of cell scratch(all P<0.01).Transwell invasion assay showed that the number of Scc-15 cells penetrating the membrane-filtered in 16 μg/ml MEL group was significantly less than that in control group ［(21.0±1.7) vs (46.0±2.3),P<0.01].Western blot results showed that compared with control group,NF-κB expression level decreased in Scc-15 cells,and expression levels of Caspase-3 and Caspase-9 increased in MEL group （all P<0.01）.
Conclusion MEL can significantly inhibit the proliferation,migration and invasion of OSCC cells and promote the apoptosis of OSCC cells,which may be related to the reduction of NF-κB production.
文章编号： 中图分类号：R739.8 文献标志码：A
基金项目:黑龙江省自然科学基金(H2018041)
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