CAJ | 학술논문

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(Chk1)小于扰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Chk1 siRNA2加浓度为(5 mg/kg)的顺铂;C组:瘤体注射顺铂(5 mg/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Chk1蛋白和mRNA的表达;运用原位缺口末端标记(TUNEL)检测各组细胞凋亡.结果 B组[(279.11±27.23) mm3]移植瘤体积明显小于C组[(512.41 ±35.18) mm3]及A组[(907.82 ±58.09) mm3],组间两两比较差异均有统计学意义(P＜0.01);B组移植瘤组织中Chk1蛋白(26.87 ±5.31)及mRNA(45.29±6.47)表达明显低于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±0.12)％],组间两两比较差异均有统计学意义(P＜0.05).结论沉默Chk1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.
목적 구건라서식관암이식류모형,관찰세포주기검측점격매1(Chk1)소우우RNA(siRNA)간우표체재체대순박유도적식관암이식류세포생물학행위적영향.방법 분조사양하류라서.A조:대조조,류체주사무균생리염수;B조:류체주사Chk1 siRNA2가농도위(5 mg/kg)적순박;C조:류체주사순박(5 mg/kg).관찰각조이식류적생장;채용면역조직화학、원위잡교방법검측각조이식류조직중Chk1단백화mRNA적표체;운용원위결구말단표기(TUNEL)검측각조세포조망.결과 B조[(279.11±27.23) mm3]이식류체적명현소우C조[(512.41 ±35.18) mm3]급A조[(907.82 ±58.09) mm3],조간량량비교차이균유통계학의의(P＜0.01);B조이식류조직중Chk1단백(26.87 ±5.31)급mRNA(45.29±6.47)표체명현저우C조(단백:56.80±7.00;mRNA:87.51 ±8.38)급A조(단백:98.23±15.67;mRNA:187.23±18.55),조간량량비교차이균유통계학의의(P＜0.05);B조세포중조망지수[(21.03±1.35)％]명현고우C조[(12.41 ±1.23)％]급A조[(1.34±0.12)％],조간량량비교차이균유통계학의의(P＜0.05).결론 침묵Chk1표체,가체내증강순박대라서식관암이식류생장억제,유도이식류세포조망적효응.
Objective Constructing the nude mouse esophageal cancer transplanted tumor model,to observe the effect of checkpoint kinase 1 (Chk1) small interfering RNA (siRNA) interference expression 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 Chk1 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 Chk1 in the tumor tissues,using in situ hybridization to test the mRNA expression of Chk1 in the tumor tissues,using TdT-mediated dUTP nick end labeling (TUNEL) method to test the apoptosis situation of tumor tissues.Results The volume of transplanted tumor in the B group [(279.11 ± 27.23) mm3] was noticeably less than that in the C group [(512.41 ±35.18) mm3] and A group [(907.82 ±58.09) mm3],there were a statistical difference among three groups (P ＜ 0.01),the expression of Chk1 protein (26.87 ± 5.31) or mRNA (45.29 ± 6.47) in B group was significantly lower than that 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 resuhs 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) ％],there were a statistical difference among three groups (P ＜ 0.05).Conclusion Silencing Chk1 expression may strengthen the growth inhibition of nude mouse esophageal cancer transplantation tumor which cisplatin bring on,induce cell apoptosis of transplantation tumor.