Yang Hua-wei, Liu Jian-lun, Liao De-zhong, Sun Yuan, Chen Zhu-shun, Zhang Xiao-li
Department of Breast Surgery, Guangxi Medical University, Nanning, China.
Zhonghua Yi Xue Za Zhi. 2012 Jul 10;92(26):1845-8.
To construct a breast cancer cell line MD-MB-231 stably overexpressing RSK4 gene and study its in vivo effects on tumor tumorigenesis.
The MD-MB-231 cells were transfected with pcDNA3.1/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.
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.
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.
构建稳定过表达RSK4基因的乳腺癌细胞系MD-MB-231，并研究其对肿瘤发生的体内影响。
分别用脂质体转染法将pcDNA3.1/Neo和pcDNA3.1/Neo-RSK4转染MD-MB-231细胞。将稳定表达RSK4（MR11和MR12）和对照载体（MN10和MN11）接种到严重联合免疫缺陷（SCID）小鼠皮下，建立SCID小鼠人乳腺癌模型。6至10周后观察异种移植瘤的生长、侵袭和转移情况。
成功筛选出稳定细胞系MR11、MR12和MN10、MN11。构建了人乳腺癌移植模型并解剖SCID小鼠。6周后，MN10/MN11组SCID小鼠皮下出现10/10个转移性肿瘤，而MR11/MR12组分别为6/10和7/10。与MN10和MN11相比，MR11和MR12的肿瘤尺寸更小，肿瘤体积和重量显著降低（P<0.001）。对照组中，80%（8/10）的小鼠发生内脏转移，而注射过表达RSK4的MDA-MB-231细胞的小鼠中40%（4/10）发生转移。苏木精和伊红染色的肺石蜡切片组织学检查显示，注射载体对照细胞的小鼠中有大量转移灶，而过表达RSK4的细胞转移灶明显减少。
SCID小鼠人乳腺癌移植与临床肿瘤患者的病程密切相关。RSK4过表达可抑制SCID小鼠人乳腺癌移植瘤的生长。
