细胞周期检测点激酶1小干扰RNA干扰表达载体体内对顺铂诱导的食管癌移植瘤细胞生长、凋亡的影响

The effect of checkpoint kinase 1 small interfering RNA on cisplatin induced esophageal carcinomatransplanted tumor cells biological behavior in vivo

目的构建裸鼠食管癌移植瘤模型，观察细胞周期检测点激酶1（Chkl）小干扰RNA（siRNA）干扰表达载体对顺铂诱导的食管癌移植瘤细胞生物学行为的影响。方法分组饲养荷瘤裸鼠。A组：对照组，瘤体注射无菌生理盐水；B组：瘤体注射ChklsiRNA2加浓度为（5mg/kg）的顺铂；C组：瘤体注射顺铂（5mg/kg）。观察各组移植瘤的生长；采用免疫组织化学、原位杂交方法检测各组移植瘤组织中Chkl蛋白和mRNA的表达；运用原位缺口末端标记（TUNEL）检测各组细胞凋广。结果B组[（279．11±27．23）mm^3]移植瘤体积明显小于C组[（512．41±35．18）mm^3]及A组[（907．82±58．09）mm^3]，组问两两比较差异均有统计学意义（P〈0．01）；B组移植瘤组织中Chkl蛋白（26．87±5．31）及mRNA（45．29±6．47）表达明碌低于C组（蛋白：56．80±7．00；mRNA：87．51±8．38）及A组（蛋白：98．23±15．67；mRNA：187．23±18．55），组间两两比较差异均有统计学意义（P〈0．05）；B组细胞中凋亡指数[（21．03±1．35）％]明显高于c组[（12．41±1．23）％]及A组[（1．34±O．12）％]，组问两两比较差异均有统计学意义（P〈0．05）。结论沉默Chkl表达，可体内增强顺铂对裸鼠食管癌移植瘤生长抑制，诱导移植瘤细胞凋亡的效应。

Objective Constructing the nude mouse esophageal cancer transplanted tumor model, to observe the effect of checkpoint kinase 1 （ Chkl ） small interfering RNA （siRNA） interference expres sion vector on cisplatin induced esophageal carcinoma transplanted tumor cells biological behavior in vivo. Methods Group breeding tumor-burdened nude mouse, namely A groups： injection Sterile Saline; B group： injection Chkl siRNA add cisplatin （5 mg/kg）, C groups： injection cisplatin （5 mg/kg）. Observing the transplantation tumor growth, using immunohistochemical to test the protein expression of Chkl in the tumor tissues, using in situ hybridization to test the mRNA expression of Chkl in the tumor tissues, using TdT-mediated dUTP nick end labeling （TUNEL） method to test the apoptosis situation of tumor tissues. Re suits The volume of transplanted tumor in the B group [ （279. 11 ± 27.23） nun3 ] was noticeably less than that in the C group [ （512. 41 ±35. 18） mm^3 ] and A group [ （907. 82 ±58. 09） mm^3 I , there were a statisti cal difference among three groups （ P 〈 0. 01 ）, the expression of Chkl protein （ 26. 87 ± 5.31 ） or mRNA （ 45.29 ± 6. 47 ） in B group was significantly lower than thal in the C group （ protein ： 56. 80 ± 7.00 ; mRNA ： 87.51 ±8.38） and A group （protein： 98.23 ± 15.67; mRNA： 187.23 ± 18.55）, there were a statistical difference among three groups （ P 〈 0. 05 ）, the results of TUNEL method detection ： the cells apoptosis index （AI） in group B [ （21.03 ± 1.35）% ] is obviously higher than that in group C [ （ 12.41 ± 1.23）% ] and group A [ （1.34 ± 0. 12 ）% 1 , there were a statistical difference among three groups （P 〈 0. 05 ）. Conclu sion Silencing Chkl expression may strengthen the growth inhibition of nude mouse esophageal cancer trans plantation tumor which cisplatin bring on, induce cell apoptosis of transplantation tumor.