CAJ | 학술논문

目的探讨静默人类真核延伸因子1A2(EEF1A2)对胰腺癌细胞体内生长的影响及其可能的机制.方法建立胰腺癌裸鼠皮下移植瘤模型,均分为空白组、阴性对照组、EEF1A2组,移植瘤内分别注射PBS、阴性对照siRNA和特异性EEF1A2 siRNA.测定各组移植瘤的体积和质量；应用免疫组织化学的方法检测各组移植瘤组织中EEF1A2和抗增殖细胞核抗原抗体(PCNA)的表达；应用TUNEL法检测各组移植瘤组织中细胞凋亡率.结果在裸鼠移植瘤模型中,从给药后第17天开始,EEF1A2组移植瘤体积增长明显滞后于阴性对照组和空白组(P值均＜0.05).治疗结束后切取肿瘤称重,EEF1A2组肿瘤质量为(0.27±0.06)g,显著低于阴性对照组和空白组[(0.39±0.08)g和(0.43±0.07)g,P＜0.05].EEF1A2主要表达于胰腺癌细胞胞质,其中在阴性对照组和空白组中,阳性细胞分布比较密集,阳性率分别为(72.58±25.47)％和(76.75±23.19)％,而EEF1A2组阳性细胞分布较稀疏,阳性率为(34.78±21.36)％,差异有统计学意义(P＜0.01).EEF1A2组的PCNA蛋白表达也显著低于空白组和阴性对照组(P＜0.01),而原位末端转移酶标记技术(TUNEL)的结果显示EEF1A2组中细胞凋亡率高于空白组和阴性对照组(P＜0.01).结论 EEF1A2在体内能被有效静默,EEF1A2静默后能明显抑制胰腺癌细胞的生长,可能与其抑制细胞增殖和促进细胞凋亡有关.
목적 탐토정묵인류진핵연신인자1A2(EEF1A2)대이선암세포체내생장적영향급기가능적궤제.방법 건립이선암라서피하이식류모형,균분위공백조、음성대조조、EEF1A2조,이식류내분별주사PBS、음성대조siRNA화특이성EEF1A2 siRNA.측정각조이식류적체적화질량；응용면역조직화학적방법검측각조이식류조직중EEF1A2화항증식세포핵항원항체(PCNA)적표체；응용TUNEL법검측각조이식류조직중세포조망솔.결과 재라서이식류모형중,종급약후제17천개시,EEF1A2조이식류체적증장명현체후우음성대조조화공백조(P치균＜0.05).치료결속후절취종류칭중,EEF1A2조종류질량위(0.27±0.06)g,현저저우음성대조조화공백조[(0.39±0.08)g화(0.43±0.07)g,P＜0.05].EEF1A2주요표체우이선암세포포질,기중재음성대조조화공백조중,양성세포분포비교밀집,양성솔분별위(72.58±25.47)％화(76.75±23.19)％,이EEF1A2조양성세포분포교희소,양성솔위(34.78±21.36)％,차이유통계학의의(P＜0.01).EEF1A2조적PCNA단백표체야현저저우공백조화음성대조조(P＜0.01),이원위말단전이매표기기술(TUNEL)적결과현시EEF1A2조중세포조망솔고우공백조화음성대조조(P＜0.01).결론 EEF1A2재체내능피유효정묵,EEF1A2정묵후능명현억제이선암세포적생장,가능여기억제세포증식화촉진세포조망유관.
Objective To explore the effect and the possible mechanisms of silent homo sapiens eukaryotic translation elongation factor 1 alpha 2(EEF1A2)gene on the growth of pancreatic cancer cell in vivo.Methods The pancreatic cancer xenograft models in mice were established.The mice were equally divided into control group,negative control group and EEF1A2 group,which were injected with PBS,negative control siRNA and EEF1A2 siRNA into xenograft tumors respectively.The size and weight of tumors in each group were measured.The expression of EEF1A2 and PCNA in tumor tissue of each group was detected by immunohistochemistry.The cell apoptosis rate in tumor tissue of each group was determined by TUNEL.Results In xenograft nude mice models,since the 17th day of injection,the growth of tumor size in EEF1A2 group was obviously slower than that of negative control group and control group(all P＜0.05).By the end of the treatment,the tumors were cut off and weighted.The weight of tumors in EEF1A2 group(0.27g± 0.06g)were significantly lower than those of control group and negative control group(0.39g± 0.08g and 0.43g± 0.07g,P＜0.05).EEF1A2 mostly expressed in cytoplasm of pancreatic cancer cell.In negative control group and control group,the positive cells distributed densely and the positive rate was(72.58 ± 25.47)％ and (76.75±23.19)％ respectively.The distribution of positive cells in EEF1A2 group was scattered and the positive rate was(34.78±21.36)％,the difference was statisically significant(P＜0.01).The expression of PCNA at protein level in EEF1A2 group was significantly lower those that of control group and negative control group(P＜ 0.01).The result of TUNEL test indicated that the cell apoptosis rate in EEF1A2 group was higher than those of control group and negative control group (P＜0.01).Conclusions The EEF1A2 gene can be effectively silented in vivo,which significantly inhibits the growth of pancreatic cancer cell.It may be related with inhibition of cell proliferation and promotion cell apoptosis.