Fan Yuanming, Wu Chengyi
Department of Endocrine Breast Surgery, The First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
Contemp Oncol (Pozn). 2013;17(5):435-9. doi: 10.5114/wo.2013.35290. Epub 2013 Oct 11.
ATP-binding cassette super family G2 (ABCG2) is recognized as the key point of the cancer cells' resistance to chemotherapy drugs. But there have been rare reports about the relationship between ABCG2 and the invasion, migration and animal tumor formation abilities of cancer cells. We want to establish a recombinant lentiviral vector with ABCG2 overexpression, and study the vector's effect on human breast cancer MCF-7 cells' biological abilities above to improve our understanding about ABCG2.
The recombinant lentiviral vector with ABCG2 overexpression was transfected into human breast cancer MCF-7 cells. The cells' abilities of migration and invasion were tested by wound healing assay, and transwell invasive assay. The MCF-7 cells infected were injected in the left back of the nude mice. In the meantime the MCF-7 cells without anything were injected in the right back of same nude mice as the control group. Western blotting was used to detect the expression of ABCG2 in MCF-7 cells infected, the transplantation tumor tissue and the control group MCF-7 cells.
The recombinant lentiviral vector with ABCG2 overexpression infected human breast cancer MCF-7 cells successfully, and the transfection efficiency was 95.4 ±2.8%. The wound line of MCF-7 cells infected healed after 48 hours, but the line of control group MCF-7 cells still existed. The number of the cells going through the membrane in infected MCF-7 cells was 78.34 ±0.25, and the number in control group MCF-7 cells was 15.28 ±0.12, p < 0.05. A much bigger transplantation tumor appeared in the MCF-7 cell infected nude mice. The expression of ABCG2 in infected MCF-7 cells and the transplantation tumor tissue was significantly higher than that in control group MCF-7 cells, p < 0.05.
We obtained ABCG2 overexpression in human breast cancer MCF-7 cells which showed the increasing migration, invasion and animal tumor formation abilities. Therefore, the results revealed that there might be a relationship between overexpression of ABCG2 and MCF-7 cells with increasing invasion, migration and animal tumor abilities.
三磷酸腺苷结合盒超家族G2(ABCG2)被认为是癌细胞对化疗药物耐药的关键因素。但关于ABCG2与癌细胞侵袭、迁移及动物肿瘤形成能力之间的关系鲜有报道。我们欲构建ABCG2过表达的重组慢病毒载体,并研究该载体对人乳腺癌MCF-7细胞上述生物学能力的影响,以增进对ABCG2的了解。
将ABCG2过表达的重组慢病毒载体转染至人乳腺癌MCF-7细胞。通过伤口愈合实验和Transwell侵袭实验检测细胞的迁移和侵袭能力。将感染的MCF-7细胞注射到裸鼠左背部。同时,将未做任何处理的MCF-7细胞注射到同一只裸鼠的右背部作为对照组。采用蛋白质免疫印迹法检测感染的MCF-7细胞、移植瘤组织及对照组MCF-7细胞中ABCG2的表达。
ABCG2过表达的重组慢病毒载体成功感染人乳腺癌MCF-7细胞,转染效率为95.4±2.8%。感染的MCF-7细胞的伤口线在48小时后愈合,但对照组MCF-7细胞的伤口线仍然存在。感染的MCF-7细胞穿过膜的细胞数为78.34±0.25,对照组MCF-7细胞为15.28±0.12,p<0.05。感染MCF-7细胞的裸鼠出现了更大的移植瘤。感染的MCF-7细胞和移植瘤组织中ABCG2的表达明显高于对照组MCF-7细胞中的表达,p<0.05。
我们在人乳腺癌MCF-7细胞中实现了ABCG2过表达,其迁移、侵袭及动物肿瘤形成能力增强。因此,结果显示ABCG2过表达与MCF-7细胞侵袭、迁移及动物肿瘤能力增强之间可能存在关联。