自然杀伤细胞对舌鳞状细胞癌裸鼠Fas蛋白表达的影响

Influence of natural killer cells on the expression of Fas protein in nude mice with tongue squamous cell carcinoma

目的探讨自然杀伤(natural killer,NK)细胞对舌鳞状细胞癌(鳞癌)裸鼠Fas蛋白表达的影响。方法 63只裸鼠随机分为对照组、非黏附NK(non-adherent NK,NA-NK)细胞组和黏附NK(adherent NK,A-NK)细胞组各21只,皮下接种人舌鳞癌Tca8113-Tb细胞株制备鳞癌裸鼠模型。接种后第3天,对照组皮下注射生理盐水0.1mL,NA-NK细胞组皮下注射NA-NK细胞0.1mL,A-NK细胞组皮下注射A-NK细胞0.1mL,均2d注射1次,共注射3次。3组分别于注射后第1、10、15、18、21、24、27天各处死3只裸鼠,取肿瘤组织,采用Western blot法检测凋亡相关蛋白Fas表达,Tunel法观察注射后第27天细胞凋亡情况。结果注射后第27天,A-NK细胞组凋亡指数[(1.76±0.83)%]高于NA-NK细胞组[(0.71±0.66)%]和对照组[(0.34±0.26)%](P<0.05),NA-NK细胞组高于对照组(P<0.05);Western blot检测结果显示,A-NK细胞组注射后第18、21、24、27天Fas蛋白(46.55±6.59、38.71±7.39、35.27±5.23、37.64±6.76)表达高于对照组(13.19±4.28、14.97±2.86、15.04±5.69、15.48±4.55)(P<0.05),注射后第18、21、27天Fas蛋白表达高于NA-NK细胞组(20.67±6.74、24.49±5.22、20.92±8.06)(P<0.05);NA-NK细胞组注射后第24天Fas蛋白(25.59±4.93)表达高于对照组(P<0.05)。结论 NK细胞可能通过提高人舌鳞癌细胞Fas蛋白表达来诱导细胞凋亡,发挥抗肿瘤作用,且A-NK细胞诱导细胞凋亡的能力强于NA-NK细胞。

Objective To observe the influence of natural killer（NK）cells on the expression of Fas protein of tongue squamous cancer cells in nude mice.Methods Sixty-three nude mice were randomly divided into control group,non-adherent NK（NA-NK）group and adherent NK（A-NK）group,with 21 mice in each group.The nude mice models of tongue squamous cell carcinoma were prepared by subcutaneous inoculation of human tongue squamous carcinoma Tca81113-Tb cell line in three groups.After inoculation for 3 days,control group,NA-NK group and A-NK group were subcutaneously injected with 0.1 mL of normal saline,0.1 mL of NA-NK cells and 0.1 mL of A-NK cells respectively once every two days,totally for 3 times.In these three groups,one nude mouse from each group was sacrificed on the1 st,10 th,15 th,18 th,21 st,24 th and 27 days respectively after injecting normal saline,NA-NK and A-NK cells to obtain tumor tissue.The apoptosis of squamous cell carcinoma on the 27 th day was observed by TUNEL staining method,and the expression of apoptosis related protein Fas was detected by Western blot method on the 1 st,10 th,15 th,18 th,21 st,24 th and 27 th days after injection.Results The apoptosis index of tumor cells was significantly higher in A-NK group（（1.76±0.83）%）than that in NA-NK group（（0.71±0.66）%）and control group（（0.34±0.26）%）（P<0.05）,and in NA-NK group than that in control group（P<0.05）on the 27 th day after injection.Western blot results showed the expressions of Fas protein were significantly higher in A-NK group（46.55±6.59,38.71±7.39,35.27±5.23,37.64±6.76）than those in control group（13.19±4.28,14.97±2.86,15.04±5.69,15.48±4.55）on the 18 th,21 st,24 th and 27 th days（P<0.05）,and in NA-NK group on the 18 th,21 st and 27 th days（20.67±6.74,24.49±5.22,20.92±8.06）（P<0.05）.The expression of Fas protein on the 24 th day was significantly higher in NA-NK group（25.59±4.93）than that in control group（P<0.05）.Conclusion The NK cell plays an anti-tumor role by increasing the expression of Fas protein of tongue squamous carcinoma Tca8113 cells to induce the cell apoptosis,and the ability of A-NK cells is stronger than that of NA-NK cells.