CAJ | 학술논문

目的探讨小分子干扰RNA(siRNA)对人乳腺癌移植瘤组织中乳腺癌特异性基因BCSG1表达的抑制作用.方法根据BCSG1的已知cDNA序列,设计并体外转录合成4条特异性siRNA,分别导入载体质粒中,用脂质体介导分别转染人乳腺癌细胞系MCF7细胞中,以无关序列转染和未转染的MCF7细胞为对照.将用上述4条特异性siRNA和无关序列转染后的细胞连同未转染的MCF7细胞(共6组细胞)注入裸鼠皮下,构建裸鼠乳腺癌移植瘤模型,分析肿瘤生长抑制情况.采用半定量逆转录聚合酶链反应和免疫组织化学SP法分别检测6组裸鼠移植瘤中BCSG1 mRNA和BCSG1蛋白的表达情况.3组人肿瘤组织(浸润性导管癌、癌旁乳腺导管增生组织和乳腺纤维腺瘤)同时用作对照.结果 BCSG1-siRNA转染的4组的抑瘤率均明显大于无关序列转染组及未转染对照组(P＜0.01);与无关序列转染组及未转染对照组相比,BCSG1-siRNA转染的4组肿瘤组织中BCSG1蛋白的表达明显减弱;未转染细胞对照组和无关序列转染组、人乳腺浸润性导管癌组均有大量BCSG1 mRNA阳性条带,而在BCSG1-siRNA转染的4组中,仅有少量BCSG1较弱的mRNA阳性条带,与未转染细胞对照组和无关序列转染组相比,差异均有统计学意义(P＜0.01).结论 BCSG1-siRNA能有效抑制肿瘤的生长,下调裸鼠乳腺癌组织中BCSG1蛋白及mRNA的表达.
목적 탐토소분자간우RNA(siRNA)대인유선암이식류조직중유선암특이성기인BCSG1표체적억제작용.방법 근거BCSG1적이지cDNA서렬,설계병체외전록합성4조특이성siRNA,분별도입재체질립중,용지질체개도분별전염인유선암세포계MCF7세포중,이무관서렬전염화미전염적MCF7세포위대조.장용상술4조특이성siRNA화무관서렬전염후적세포련동미전염적MCF7세포(공6조세포)주입라서피하,구건라서유선암이식류모형,분석종류생장억제정황.채용반정량역전록취합매련반응화면역조직화학SP법분별검측6조라서이식류중BCSG1 mRNA화BCSG1단백적표체정황.3조인종류조직(침윤성도관암、암방유선도관증생조직화유선섬유선류)동시용작대조.결과 BCSG1-siRNA전염적4조적억류솔균명현대우무관서렬전염조급미전염대조조(P＜0.01);여무관서렬전염조급미전염대조조상비,BCSG1-siRNA전염적4조종류조직중BCSG1단백적표체명현감약;미전염세포대조조화무관서렬전염조、인유선침윤성도관암조균유대량BCSG1 mRNA양성조대,이재BCSG1-siRNA전염적4조중,부유소량BCSG1교약적mRNA양성조대,여미전염세포대조조화무관서렬전염조상비,차이균유통계학의의(P＜0.01).결론 BCSG1-siRNA능유효억제종류적생장,하조라서유선암조직중BCSG1단백급mRNA적표체.
Objective To investigate the inhibitory effects of siRNA targeting BCSG1 gene expression in tumor transplants of human breast cancer cell line in nude mice. Methods Four-pairs of small interfering RNA sequences of BCSG1 were chemically synthesized and inserted into the plasmid expression vectors, and were then transfected into human breast carcinoma cell line MCF7 by liposome method. Plasmid vector with unrelated sequence was used as the vector control. Cells transfected with 4siRNA sequences, control vector and naive FCF7 cells were transplanted into the nude mice. The tumor inhibition was analysised. Immunohistochemical SP method and semi-quantitative RT-PCR were adopted to detect the BCSG1 mRNA and protein expression, respectively. Breast tissue samples of human infiltrating ductal carcinoma, ductal hyperplasia and fibroadenoma were also used as the controls. Results The inhibition rates of tumor growth in four BCSG1-siRNA transfected groups were remarkably higher than those of the vector control group and naive MCF7 cells (P ＜0. 01 ). Compared with that of the vector control and na(i)ve MCF7 cell group, there was a significant decrease of BCSG-1 protein expression in the four experimental groups by immuno-histochemistry staining (P ＜0. 01 ). In addition, BCSG1 mRNA expression in the four groups transfected with BCSG1-siRNA were significantly less than that of the control vector group,naive MCF7 cell control group and human breast IDC (P ＜ 0. 01 ). Conclusion BCSG1-siRNA downregulates the expression of BCSG1 and inhibits effectively growth of the transplaned human breast cancer cell line in nude mice.