CAJ | 학술논문

目的探讨短发夹RNA(shRNA)沉默CXCR4基因对小鼠黑素瘤的抑瘤效应及器官转移的影响.方法设计合成CXCR4特异性shRNA插入pSilencer载体,转染人高转移性黑素瘤细胞株MV3,通过小鼠尾静脉瘤细胞注射方法,建立黑素瘤转移模型,观察肿瘤生长情况,分析CXCR4基因沉默对黑素瘤细胞肝、脑、肺等实质性器官转移的影响.实验随机分为3组:A组(CXCR4-shRNA转染组)、B组(无关对照组)和C组(空白对照组).结果小鼠黑素瘤皮下种植瘤成功率为100％,肿瘤生长曲线表明,A组肿瘤生长明显抑制.A组与C组和B组比较差异具有统计学意义(P＜0.01).肝、脑组织内转移瘤数目A组较B组和C组明显少.黑素瘤定向肺转移能力下降:B、C组和A组肺转移发生率分别为90.0％、87.5％和20.0％；B、C组和A组的肺转移结节数目分别为206、165和23个.结论 shRNA介导的CXCR4基因沉默可在黑素瘤皮下种植瘤产生抑瘤效应,明显降低肝、脑器官转移能力和定向肺转移能力.
목적 탐토단발협RNA(shRNA)침묵CXCR4기인대소서흑소류적억류효응급기관전이적영향.방법 설계합성CXCR4특이성shRNA삽입pSilencer재체,전염인고전이성흑소류세포주MV3,통과소서미정맥류세포주사방법,건립흑소류전이모형,관찰종류생장정황,분석CXCR4기인침묵대흑소류세포간、뇌、폐등실질성기관전이적영향.실험수궤분위3조:A조(CXCR4-shRNA전염조)、B조(무관대조조)화C조(공백대조조).결과 소서흑소류피하충식류성공솔위100％,종류생장곡선표명,A조종류생장명현억제.A조여C조화B조비교차이구유통계학의의(P＜0.01).간、뇌조직내전이류수목A조교B조화C조명현소.흑소류정향폐전이능력하강:B、C조화A조폐전이발생솔분별위90.0％、87.5％화20.0％；B、C조화A조적폐전이결절수목분별위206、165화23개.결론 shRNA개도적CXCR4기인침묵가재흑소류피하충식류산생억류효응,명현강저간、뇌기관전이능력화정향폐전이능력.
Objective To study the inhibiting effects on the invasion and metastasis of melanoma by CXCR4 gene silence in nude mice.Methods The CXCR4 specific recombinant plasmid vector was constructed and transfected into the cultured MV3 cell line with lipofectamine.The models of subcutaneous melanoma in nude mice were established with MV3 cells.The nude mouse model of lung metastasis was established by injection of MV3 cells into the tail vein.The animals were sacrificed at 8weeks after the melanoma cells injection.CXCR4-shRNA plasmid vectors were discontinuously injected directly into the established tumor and vein.The changes of weight and size of the tumors and the mice body weight during the therapy were calculated respectively.Histological observation was performed to evaluate the presence and number of metastatic tumors.Results The subcutaneous melanoma tumors in nude mice were established successfully.The growth of tumors in the CXCR4-shRNA injected nude mice was inhibitted obviously through tumor growth curve. There were significant differences between negative shRNA control nude mice and blank control nude mice (P＜0.01).Melanoma cells with CXCR4 shRNA permanent transfection had a much lower lung and brain and liver metastatic potential in nude mice than control cells and mock control cells in vivo.Conclusions CXCR4 gene silencing mediated by shRNA significantly suppresses the growth of MV3 cell in vitro.Silencing of CXCR4 mediated by shRNA can also effectively decrease the metastatic potential of lung and liver and brain.