CAJ | 학술논문

目的观察E2F-1基因沉默的慢病毒载体E2F-1/RNAi-LV对人胃癌裸鼠皮下瘤耐药性的影响.方法将人胃癌SGC7901-顺铂(DDP)耐药细胞接种于裸鼠皮下,建立人胃癌耐药裸鼠皮下瘤模型.将成瘤的36只裸鼠随机分为E2F-1/RNAi-LV组、LV-scrRNAi组和PBS组,每组12只.隔日向3组裸鼠肿瘤内分别注射E2F-1/RNAi-LV、LV-scrRNAi或PBS,并按体重腹腔注射DDP,共9次.注射期间和处死裸鼠后测量各组裸鼠的肿瘤体积.应用TUNEL法检测肿瘤细胞的凋亡指数,应用半定量RT-PCR和Western blot法检测肿瘤组织中E2F-1、c-Myc、survivin、多药耐药基因1(MDR1)和多药耐药相关蛋白(MRP) mRNA和蛋白的表达.结果注射慢病毒后,E2F-1/RNAi-LV组肿瘤的生长速度明显减慢.处死裸鼠后,E2F-1/RNAi-LV组裸鼠的肿瘤体积为(745.13±154.42)mm3,明显低于LV-scrRNAi组[(1988.80±171.35) mm3]和PBS组[(1633.48±215.25) mm3,P＜0.05].E2F-1/RNAi-LV组肿瘤组织的凋亡指数为(27.5±9.7)％,明显高于LV-scrRNAi组[(7.0±1.1)％]和PBS组[(7.3±1.2)％,P＜0.05].E2F-1/RNAi-LV组肿瘤组织中E2F-1 mRNA和蛋白的表达量分别为0.40±0.32和0.35±0.01,明显低于LV-scrRNAi组和PBS组(P＜0.05).E2F-1/RNAi-LV组c-Myc、survivin、MDR1和MRP mRNA和蛋白的表达量亦明显低于LV-scrRNAi组和PBS组(均P＜0.05).结论慢病毒载体E2F-1/RNAi-LV瘤内注射可降低人胃癌SGC7901-DDP耐药细胞对DDP的耐药性,抑制人胃癌耐药裸鼠皮下瘤的生长.
목적 관찰E2F-1기인침묵적만병독재체E2F-1/RNAi-LV대인위암라서피하류내약성적영향.방법 장인위암SGC7901-순박(DDP)내약세포접충우라서피하,건립인위암내약라서피하류모형.장성류적36지라서수궤분위E2F-1/RNAi-LV조、LV-scrRNAi조화PBS조,매조12지.격일향3조라서종류내분별주사E2F-1/RNAi-LV、LV-scrRNAi혹PBS,병안체중복강주사DDP,공9차.주사기간화처사라서후측량각조라서적종류체적.응용TUNEL법검측종류세포적조망지수,응용반정량RT-PCR화Western blot법검측종류조직중E2F-1、c-Myc、survivin、다약내약기인1(MDR1)화다약내약상관단백(MRP) mRNA화단백적표체.결과 주사만병독후,E2F-1/RNAi-LV조종류적생장속도명현감만.처사라서후,E2F-1/RNAi-LV조라서적종류체적위(745.13±154.42)mm3,명현저우LV-scrRNAi조[(1988.80±171.35) mm3]화PBS조[(1633.48±215.25) mm3,P＜0.05].E2F-1/RNAi-LV조종류조직적조망지수위(27.5±9.7)％,명현고우LV-scrRNAi조[(7.0±1.1)％]화PBS조[(7.3±1.2)％,P＜0.05].E2F-1/RNAi-LV조종류조직중E2F-1 mRNA화단백적표체량분별위0.40±0.32화0.35±0.01,명현저우LV-scrRNAi조화PBS조(P＜0.05).E2F-1/RNAi-LV조c-Myc、survivin、MDR1화MRP mRNA화단백적표체량역명현저우LV-scrRNAi조화PBS조(균P＜0.05).결론 만병독재체E2F-1/RNAi-LV류내주사가강저인위암SGC7901-DDP내약세포대DDP적내약성,억제인위암내약라서피하류적생장.
Objective To study the effects of E2F-1-silencing lentivirus vector on the growth and chemoresistance of subcutaneous human gastric cancer in nude mice.Methods Thirty-six nude mice were inoculated subcutaneously with chemoresistant SGC-7901/DDP cells to establish subcutaneous tumor models of gastric carcinoma.The mice were randomly divided into E2F-1/RNAi-LV group,LV-scrRNAi group and PBS group (n =12).E2F-1/RNAi-LV,LV-scrRNAi or PBS (0.1 ml per time) was injected into the mice,respectively,every two days.The nude mice received an intraperitoneal injection of cisplatin (25 mg/kg)every two days.The tumor volume was measured and histopathologieal changes of the tumors were observed by HE staining.The expressions of E2F-1,c-Myc,survivin,MDR1 and MRP were detected by semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot.Apoptosis in tumor xenografts was determined by in situ TUNEL labeling technique.Results The mean tumor growth rate of the E2F-1/RNAi-LV group was significantly slower than that of the LV-scrRNAi and control groups (P ＜0.05).The tumor volume of the E2F-1/RNAi-LV group was (745.13 ± 154.42) mm3,significantly lower than that of the LV-scrRNAi and PBS groups (P ＜ 0.05).Compared with that in the LV-scrRNAi and PBS groups,the expressions of mRNA and protein of E2F-1,c-Myc,survivin,MDR1 and MRP were significantly decreased in the E2F-1/RNAi-LV group (P ＜0.05).The apoptotic rate in the E2F-1/RNAi-LV treatment group was (27.5 ± 9.7) ％,significantly higher than (7.0 ± 1.1) ％ in the LV-scrRNAi group and (7.3 ± 1.2) ％ in the PBS group (P ＜ 0.05).Conclusion Intra-tumoral injection of E2F-1/RNAi-LV shows significantly inhibitory effect on the tumor growth and chemoresistance of subcutaneous human gastric cancer in nude mice.