CAJ | 학술논문

目的探讨慢病毒载体介导RNA干扰沉默Livin基因表达,对大肠癌HT-29细胞裸鼠移植瘤生长及放射敏感性的影响.方法选取36只BALB/c(nu/nu)裸鼠,按随机区组法分为空白对照组、阴性对照组及实验组,将HT-29细胞接种于裸鼠,建立皮下移植瘤裸鼠模型,观察裸鼠体重及瘤体积的变化.RT-PCR、免疫组织化学分析Livin mRNA及蛋白表达的变化,原位末端标记法(TUNEL)法检测细胞凋亡的变化;瘤内注射生理盐水、空载慢病毒和干扰慢病毒,同时均给予10 Gy 6MVX射线照射,绘制裸鼠体重和移植瘤生长曲线.结果实验组体积抑瘤率为(50.04±0.07)％,瘤体重量明显减轻(F=4.85,P＜0.05),瘤重抑瘤率为(50.27 ±0.17)％.实验组Livin mRNA表达水平为(17.75±0.08)％,明显低于空白对照组的(67.60±0.05)％和阴性对照组的(68.54±0.03)％(F=89.97,P＜0.01).实验组Livin蛋白表达水平为(36.00±3.40)％,明显低于空白对照组的(85.00±3.15)％和阴性对照组的(80.33±3.08)％(F=107.32,P＜0.01).实验组凋亡率为(23.67±2.25)％,明显高于空白对照组的(5.00±1.50)％和阴性对照组的(8.33±1.82)％(F=56.94,P＜0.01).与射线联合时,裸鼠移植瘤体积在分组之间总体存在差异(F=10.70,P＜0.01),实验组肿瘤体积变化小于阴性对照组和空白对照组(F=7.01 ～9.32,P＜0.01).结论慢病毒载体介导RNA干扰沉默Livin基因表达能抑制大肠癌HT-29细胞裸鼠移植瘤的生长,并增加移植瘤对放疗的敏感性.
목적 탐토만병독재체개도RNA간우침묵Livin기인표체,대대장암HT-29세포라서이식류생장급방사민감성적영향.방법 선취36지BALB/c(nu/nu)라서,안수궤구조법분위공백대조조、음성대조조급실험조,장HT-29세포접충우라서,건립피하이식류라서모형,관찰라서체중급류체적적변화.RT-PCR、면역조직화학분석Livin mRNA급단백표체적변화,원위말단표기법(TUNEL)법검측세포조망적변화;류내주사생리염수、공재만병독화간우만병독,동시균급여10 Gy 6MVX사선조사,회제라서체중화이식류생장곡선.결과 실험조체적억류솔위(50.04±0.07)％,류체중량명현감경(F=4.85,P＜0.05),류중억류솔위(50.27 ±0.17)％.실험조Livin mRNA표체수평위(17.75±0.08)％,명현저우공백대조조적(67.60±0.05)％화음성대조조적(68.54±0.03)％(F=89.97,P＜0.01).실험조Livin단백표체수평위(36.00±3.40)％,명현저우공백대조조적(85.00±3.15)％화음성대조조적(80.33±3.08)％(F=107.32,P＜0.01).실험조조망솔위(23.67±2.25)％,명현고우공백대조조적(5.00±1.50)％화음성대조조적(8.33±1.82)％(F=56.94,P＜0.01).여사선연합시,라서이식류체적재분조지간총체존재차이(F=10.70,P＜0.01),실험조종류체적변화소우음성대조조화공백대조조(F=7.01 ～9.32,P＜0.01).결론 만병독재체개도RNA간우침묵Livin기인표체능억제대장암HT-29세포라서이식류적생장,병증가이식류대방료적민감성.
Objective To explore the effects of silencing Livin gene by RNA interference mediated by lentiviral vector on colorectal cancer HT-29 cell xenograft growth and sensitivity to radiotherapy in nude mice.Methods BALB/c nude mice models were established by subcutaneously inoculating differently treated HT-29 cells into nude mice and the tumor growth situation of tumors was observed by measuring the volume of tumors and the weight of the nude mice at different time points after cell seeding.Livin expression was detected by RT-PCR and immunohistochemistry,respetively.Apoptosis rate was detected by TUNEL.Normal saline,lentivirus carring unrelated sequences,lentivirus caning Livin shRNA were injected intratumorally.All the nude mice were given 10 Gy of 6 MV X-ray irradiation.The changes of mice weight and the tumor volume were measured at different time points and the weight and tumor growth curves were drawn.Results The inhibition rate of tumor volume was(50.04 ± 0.07)％ and the tumor weight of the RNA interfering group was significantly less than that in experimental group compared to the blank and negative groups(F=4.85,P＜0.05),and the inhibition rate of tumor weight was(50.27 ±0.17)％.Relative Livin mRNA expression level in the RNA interfering experimental group was(17.75 ±0.08)％,and was significantly lower than that of the blank group(67.60 ± 0.05)％ and the negative group(68.54 ± 0.03)％(F=89.97,P＜0.01).Livin protein expression level in the RNA inferring group was also significantly lower[(36.00 ± 3.40)％ versus(85.00 ± 3.15)％,(80.33 ± 3.08)％,F=107.32,P＜0.01].The apoptosis rate in the RNA interfering experimental group was significantly higher than that in the blank and the negative groups[(23.67 ± 2.25)％ versus(5.00 ± 1.50)％,(8.33 ± 1.82)％,F=56.94,P＜0.01].Combined with radiotherapy,the tumor volume at different groups had significant difference(F=10.70,P＜0.01),and RNA interfering group was significantly less than negative group and blank group(F=7.01-9.32,P＜0.01).Conclusions Silencing of Livin gene expression by lentiviral vector-mediated RNA interference could inhibit the growth of colorectal HT-29 cell xenograft and increase the sensitivity of the transplanted tumors to radiotherapy.