紫甘蓝提取物对X线照射后舌癌细胞双向作用的实验研究

Experimental study of bidirectional effect of purple cabbage extract on tongue cancer cells after X-ray irradiation

目的:探讨紫甘蓝提取物(purple cabbage extract,PCE)对X线照射后舌癌细胞的双向作用。方法:将对数生长期的人舌鳞状细胞癌Tca8113细胞及人正常口腔黏膜HOK细胞分为Tca8113细胞对照组(Tca CON)、Tca8113细胞10μmol/L PCE组(Tca PCE-10μmol/L)、Tca8113细胞50μmol/L PCE组(Tca PCE-50μmol/L)、Tca8113细胞照射组(Tca IR)、Tca8113细胞照射加10μmol/L PCE组(Tca IR+PCE-10μmol/L)、Tca8113细胞照射加50μmol/L PCE组(Tca IR+PCE-50μmol/L)及HOK细胞对照组(HOK CON)、HOK细胞10μmol/L PCE组(HOK PCE-10μmol/L)、HOK细胞50μmol/L PCE组(HOK PCE-50μmol/L)、HOK细胞照射组(HOK IR)、HOK细胞照射加10μmol/L PCE组(HOK IR+PCE-10μmol/L)、HOK细胞照射加50μmol/L PCE组(HOK IR+PCE-50μmol/L)。MTT实验检测加入不同浓度PCE对舌癌细胞Tca8113及正常口腔黏膜HOK细胞增殖的影响;流式细胞术检测6 MeV X线直线加速器照射后(6 Gy)PCE对Tca8113及HOK细胞凋亡的影响;qRT-PCR实验检测在加入不同浓度PCE作用下对照射后Caspase 3及Caspase 9表达的影响。结果:MTT结果显示,PCE可明显抑制Tca8113细胞的增殖,其增殖抑制率随药物浓度的增加而升高,PCE对正常口腔黏膜HOK细胞增殖无明显的抑制率。流式细胞术结果显示,对于Tca8113细胞,PCE显著提高了照射后Tca8113细胞凋亡率(P<0.05);但对于HOK细胞,PCE显著降低了照射后HOK细胞凋亡率(P<0.05)。qRT-PCR结果显示,对于Tca8113细胞,照射给药组(加10μmol/L PCE组、加50μmol/L PCE组)Caspase 3及Caspase 9的表达较单纯照射组明显上调(P<0.05);对于HOK细胞,照射给药组(加10μmol/L PCE组、加50μmol/L PCE组)Caspase 3及Caspase 9的表达较单纯照射组明显下调(P<0.05)。结论:PCE通过调节凋亡通路对舌癌Tca8113细胞起到放射增敏的作用,对正常口腔黏膜细胞起到放射防护作用,即PCE对X线照射后舌癌细胞起到双向作用。

Objective:To investigate the bidirectional effect of purple cabbage extract(PCE)on tongue cancer cells after X-ray irradiation.Methods:The logarithmic growth of human tongue squamous cell cancer Tca8113 cells and human normal oral mucosal HOK cells were divided into Tca8113 cell control group(Tca CON),Tca8113 cell with 10μmol/L PCE group(Tca PCE-10μmol/L),Tca8113 cell with 50μmol/L PCE group(Tca PCE-50μmol/L),Tca8113 cell under irradiation(IR)group(Tca IR),Tca8113 cell under IR with 10μmol/L PCE group(Tca IR+PCE-10μmol/L),Tca8113 cell under IR with 50μmol/L PCE group(Tca IR+PCE-50μmol/L)and HOK cell control group(HOK CON),HOK cell with 10μmol/L PCE group(HOK PCE-10μmol/L),HOK cell with 50μmol/L PCE group(HOK PCE-50μmol/L),HOK cell under IR group(HOK IR),HOK cell under IR with 10μmol/L PCE group(HOK IR+PCE-10μmol/L),HOK cells under IR with 50μmol/L PCE group(HOK IR+PCE-50μmol/L).The activities of the tongue cancer cell line Tca8113 and normal oral mucosal cell line HOK with different concentrations of PCE were detected by MTT assay.The apoptosis of Tca8113 and HOK cells with different concentrations of PCE was detected by flow cytometry under IR(6 Gy).The expression of Caspase 3 and Caspase 9 with different concentrations of PCE was detected by qRT-PCR under IR.Results:MTT results showed the proliferation of Tca8113 cells was significantly inhibited by PCE in a concentration-dependent manner.However,there was no significant difference in inhibition rate on normal oral mucosal HOK cells.The results of flow cytometry showed the apoptosis rate of Tca8113 cells significantly increased under IR with PCE(P<0.05).However,for HOK cells,the result showed the apoptosis rate of HOK cells significantly decreased with PCE under IR(P<0.05).The results of qRT-PCR showed for Tca8113 cells,the expression of Caspase 3 and Caspase 9 in IR+administration group(with 10μmol/L PCE group and 50μmol/L PCE group)was significantly up-regulated than that in Tca IR group(P<0.05).For HOK cells,the expression of Caspase 3 and Caspase 9 in IR+administration group(with 10μmol/L PCE group and 50μmol/L PCE group)was significantly down-regulated than that in HOK IR group(P<0.05).Conclusion:PCE has the increased radiosensitizing effect on tongue cancer Tca8113 cells,and protecting effect for the normal oral mucosal HOK cells by regulating the apoptotic pathway,that is,PCE has the bidirectional effect in tongue cancer radiotherapy.