CAJ | 학술논문

目的构建稳定过表达RSK4基因的MD-MB-231乳腺癌细胞株,观察过表达RSK4基因对乳腺癌体内成瘤的影响.方法将pcDNA3.1/Neo和pcDNA3.1/Neo-RSK4分别转染进人乳腺癌细胞MD-MB-231,筛选出稳定的细胞株,命名为空载体组(MN10,MN11)和转染组(MR11,MR12)并种植至免疫缺陷小鼠(SCID)皮下组织,建立SCID人乳腺癌MD-MB-231细胞移植瘤模型,种植后6～10周观察移植瘤的生长、侵袭转移的情况.结果筛选出稳定表达的乳腺癌细胞株MR11和MR12和空白对照组MN10和MN11;构建SCID的人乳腺癌移植瘤模型,解剖鼠发现:注射MN10和MN11对照组细胞后6周,SCID鼠均有转移瘤生成10/10,而注射MR11和MR12细胞组后在第6周和7周分别有6/10和7/10鼠成瘤,注射MR11或MR12细胞的鼠在所形成的肿瘤大小和重量上明显小于MN10和MN11对照组；注射10周后,RSK4高表达的MR11和MR12组有4只小鼠发生内脏转移瘤,而对照组有8只小鼠出现内脏转移瘤,HE染色发现发生在肺的转移灶MN10和MN11组明显多于MR11和MR12组.结论成功构建SCID人乳腺癌MD-MB-231细胞转移瘤模型,较真实模拟人乳腺癌生长过程,证实过表达RSK4基因在体内可抑制乳腺癌生长.
목적 구건은정과표체RSK4기인적MD-MB-231유선암세포주,관찰과표체RSK4기인대유선암체내성류적영향.방법 장pcDNA3.1/Neo화pcDNA3.1/Neo-RSK4분별전염진인유선암세포MD-MB-231,사선출은정적세포주,명명위공재체조(MN10,MN11)화전염조(MR11,MR12)병충식지면역결함소서(SCID)피하조직,건립SCID인유선암MD-MB-231세포이식류모형,충식후6～10주관찰이식류적생장、침습전이적정황.결과 사선출은정표체적유선암세포주MR11화MR12화공백대조조MN10화MN11;구건SCID적인유선암이식류모형,해부서발현:주사MN10화MN11대조조세포후6주,SCID서균유전이류생성10/10,이주사MR11화MR12세포조후재제6주화7주분별유6/10화7/10서성류,주사MR11혹MR12세포적서재소형성적종류대소화중량상명현소우MN10화MN11대조조；주사10주후,RSK4고표체적MR11화MR12조유4지소서발생내장전이류,이대조조유8지소서출현내장전이류,HE염색발현발생재폐적전이조MN10화MN11조명현다우MR11화MR12조.결론 성공구건SCID인유선암MD-MB-231세포전이류모형,교진실모의인유선암생장과정,증실과표체RSK4기인재체내가억제유선암생장.
Objective To construct a breast cancer cell line MD-MB-231 stably overexpressing RSK4 gene and study its in vivo effects on tumor tumorigenesis.Methods The MD-MB-231 cells were transfected with pcDNA 3.1/Neo and pcDNA3.1/Neo-RSK4 by lipofectamin transfection respectively.The stable expression of RSK4 (MR11 and MR12) and control vector (MN10 and MN11) were inoculated into severe combined immunodeficiency (SCID) mice subcutis to establish a model of human breast cancer in SCID mice.The xenograft tumor growth,invasion and metastasis were observed after 6-10 weeks.Results The stable cell lines MR11,MR12 and MN10,MN11 were screened successfully.We constructed the human breast cancer transplanted model and dissected SCID mice.After 6 weeks,SCID mice subcutis of the MN10/MN11 group yielded 10/10 metastatic tumors versus 6/10 and 7/10 in the MR11/MR12 group respectively.MR11 and MR12 showed much smaller tumor sizes and significantly reduced tumor volume and weight versus MN10 and MN11 (P ＜0.001 ).In the control group,visceral metastasis developed in 80％(8/10) of mice while in metastasis developed in 40％ (4/10) of mice injected with RSK4-overexpressing MDA-MB-231 cells.Histological examination of hematoxylin and eosin-stained paraffin sections of lungs revealed numerous metastases in mice injected with vector control cells whereas RSK4-overexpressing cells showed markedly decreased metastatic lesions.Conclusion Transplanted human breast cancer in SCID mice closely correlates with the disease course of clinical tumor patients.Overexpression of RSK4 can inhibit tumor growth of transplanted human breast cancer in SCID mice.