CAJ | 학술논문

目的观察c-myc小干扰RNA(siRNA)联合氟尿嘧啶(5-Fu)抑制喉癌Hep-2细胞生长的体内外作用,探讨在喉癌治疗中将c-myc作为基因治疗靶点的价值.方法利用RNA干扰技术将c-myc siRNA转染到喉癌Hep-2细胞中,Western blot法检测c-myc蛋白的表达,流式细胞仪检测c-myc siRNA与5-Fu单独或联合应用,对喉癌细胞周期的影响.构建裸鼠移植瘤模型,观察c-myc siRNA及联合5-Fu对肿瘤生长的影响.免疫组织化学法检测c-myc在肿瘤组织中的表达;脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal-deoxynucleotidyl transferase mediated nick end labeling,TUNEL)检测肿瘤组织细胞的凋亡情况.结果在转染c-myc siRNA后的喉痛Hep-2细胞中,c-myc 蛋白的表达水平逐渐下调;G0-G1期的细胞开始增加,同时S期的细胞逐渐减少;当转染c-myc siRNA 细胞联合使用5-Fu时,G0-G1期的细胞明显增加,为(79.3.±2.1)%,同时S期的细胞减少为(17.5±1.7)%.c-myc siRNA联合5-Fu组移植瘤生长最慢,与c-myc siRNA组和5-Fu组相比差异有统计学意义(t值分别为44.170和78.988,P＜0.05);c-myc siRNA+5-Fu组治疗的移植瘤重量明显小于对照组,也小于c-myc siRNA组和5-Fu组(P值均＜0.05).免疫组织化学结果显示,c-myc siRNA及其联合5-Fu组肿瘤组织中c-myc蛋白的表达下调,肿瘤细胞的凋亡数明显增加高于单用5-Fu组(t=-17.871,P＜0.05).结论 c-myc siRNA可特异性地下调Hep-2细胞和裸鼠组织中c-myc的表达,联合5-Fu时,可显著抑制喉癌移植瘤的生长,促进肿瘤细胞的凋亡,表明c-myc可能是喉癌基因治疗中一个重要的分子靶点.
목적 관찰c-myc소간우RNA(siRNA)연합불뇨밀정(5-Fu)억제후암Hep-2세포생장적체내외작용,탐토재후암치료중장c-myc작위기인치료파점적개치.방법 이용RNA간우기술장c-myc siRNA전염도후암Hep-2세포중,Western blot법검측c-myc단백적표체,류식세포의검측c-myc siRNA여5-Fu단독혹연합응용,대후암세포주기적영향.구건라서이식류모형,관찰c-myc siRNA급연합5-Fu대종류생장적영향.면역조직화학법검측c-myc재종류조직중적표체;탈양핵당핵감산말단전이매개도적결구말단표기법(terminal-deoxynucleotidyl transferase mediated nick end labeling,TUNEL)검측종류조직세포적조망정황.결과 재전염c-myc siRNA후적후통Hep-2세포중,c-myc 단백적표체수평축점하조;G0-G1기적세포개시증가,동시S기적세포축점감소;당전염c-myc siRNA 세포연합사용5-Fu시,G0-G1기적세포명현증가,위(79.3.±2.1)%,동시S기적세포감소위(17.5±1.7)%.c-myc siRNA연합5-Fu조이식류생장최만,여c-myc siRNA조화5-Fu조상비차이유통계학의의(t치분별위44.170화78.988,P＜0.05);c-myc siRNA+5-Fu조치료적이식류중량명현소우대조조,야소우c-myc siRNA조화5-Fu조(P치균＜0.05).면역조직화학결과현시,c-myc siRNA급기연합5-Fu조종류조직중c-myc단백적표체하조,종류세포적조망수명현증가고우단용5-Fu조(t=-17.871,P＜0.05).결론 c-myc siRNA가특이성지하조Hep-2세포화라서조직중c-myc적표체,연합5-Fu시,가현저억제후암이식류적생장,촉진종류세포적조망,표명c-myc가능시후암기인치료중일개중요적분자파점.
Objective To observe the effects of small interfere RNA (siRNA) targeting the c-myc in combination with 5-fluorouracil (5-Fu) on the growth of Hep-2 cells in vitro and in vivo.Methods Hep-2 cells transfected with or without c-myc siRNA were treated with 5-Fu for 48 h.C-myc protein levels in Hep-2 cells were detected using the Western blot.The cell cycle was analyzed by flow cytometry.Hep-2 cells were subcutaneously inoculated into the back of BALB/c nude mice to establish the implanted laryngeal squamous carcinoma model.PBS,c-myc siRNA,and 5-Fu,alone or in combinations were administered i.p.The mice were sacrificed after the treatments and the tumor masses were removed to determine the tumor volume and weight.The inhibitory rate was calculated.Expression of c-myc in tumor tissue was detected by immunocytochemistry and cell apoptosis was analyzed by terminal transferase dUTP nick end labeling (TUNEL).Results The protein levels of c-myc decreased after transfected with c-myc siRNA.C-myc siRNA-transfected cells showed an increase in the percentage of cells in the G0-G1 phase and a decrease in the percentage of cells in the S phase.When combined with 5-Fu,the results were improved.The tumor growth was faster in the control group and was significantly slower in the c-myc siRNA plus 5-Fu group than that in the c-myc siRNA group or 5-Fu group ( P ＜0.05 ) .The tumor weight in the c-myc siRNA plus 5-Fu group was significantly smaller than that in the c-myc siRNA or 5-Fu group ( P ＜ 0.05 ).Immunohistochemistry showed that c-myc siRNA inhibited the expression of c-myc in tumor tissues in the c-myc siRNA group and c-myc siRNA plus 5-Fu group (P ＜0.05).The number of apoptotic cells in the c-myc siRNA plus 5-Fu group was higher than those in the c-myc siRNA groups (P ＜0.05).Conclusions C-myc siRNA inhibits the expression of c-myc in Hep-2 cells and in the tumor tissues of nude mice.C-myc siRNA combined with 5-Fu inhibits the growth of implanted laryngeal squamous carcinoma and promotes cell apoptosis.C-myc could become a novel target for the treatment of laryngeal squamous carcinoma.