骨桥蛋白反义基因对乳腺癌荷瘤裸鼠肿瘤生长及肺转移的影响

Effect of Anti-sense Osteopontin Gene on Growth and Pulmonary Metastasis of Tumor in Nude Mice with Breast Cancer

目的探讨骨桥蛋白反义基因(ANOPN)对乳腺癌荷瘤裸鼠肿瘤生长及肺转移的影响。方法构建人骨桥蛋白反义基因真核表达质粒pcDNA3.1-ANOPN,将质粒pcDNA3.1-ANOPN、空载体pcDNA3.1(+)和等量的脂质体分别转染人乳腺癌细胞系MDA-MA-231,将转染细胞分别命名为MDA-ANOPN、MDA-vect和MDA,接种至裸鼠,观察肿瘤的生长、转移情况,并用免疫组化法检测肿瘤组织切片中OPN的表达。结果重组质粒pcDNA3.1-ANOPN经酶切、PCR及测序,证明构建正确。MDA-ANOPN组裸鼠出瘤时间、肿瘤体积及重量较MDA-vect和MDA组均显著降低,肿瘤肺转移灶数量减少,OPN表达较低。结论ANOPN可抑制裸鼠移植瘤的生长及肺转移。

Objective To study the effect of anti-sense osteopontin （ANOPN） gene on growth and pulmonary metastasis of tumor in nude mice with breast cancer. Methods Eukaryotic expression vector pcDNA3. 1-ANOPN was constructed and used to transfect human breast cancer cell line MDA-MB-231, using empty vector pcDNA3. 1 （＋） and liposome as parallel and blank controls respectively. The MDA-MB-231 cells transfected with pcDNA3. 1-ANOPN, pcDNA3. 1 （＋） and liposome, named as MDA-ANOPN, MDA-vect and MDA respectively, were inoculated to nude mice, and the growth and metastasis of tumor were observed. The expression of OPN in section of tumor tissue was determined by immunohistochemical method. Results Restriction analysis, PCR and sequencing proved that recombinant plasmid pcDNA3. 1-ANOPN was constructed correctly. Compared with those in MDA-vect and MDA groups, the time when tumor appeared in MDA-ANOPN group was shortened significantly, while size and weight of tumors decreased significantly, and the number of pulmonary metastatic foci as well as expression level of OPN decreased. Conclusion ANOPN inhibited the growth and pulmonary metastasis of transplanted tumor in nude mice.