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背景与目的近年来,肺癌分子靶向治疗取得了明显进展,其中抗血管生成分子靶向治疗已引起人们广泛关注。本研究的目的是探讨经人血管能抑素(canstatin)基因重组表达载体转染的人淋巴细胞培养上清对动物肿瘤生长和转移的抑制作用。方法将canstatin重组表达载体及空载体通过电穿孔的方法转染人淋巴细胞,行G418筛选获得转基因细胞克隆;用SDS-PAGE电泳检测canstatin蛋白在转基因细胞培养上清中的表达。将Lewis肺癌细胞接种于C57BL小鼠皮下,成瘤后将30只小鼠随机分成3组,分别给予重组载体转染的淋巴细胞培养上清、空载体转染的淋巴细胞培养上清和生理盐水(NS)各0.2mL在腹股沟皮下注射,每天1次,连续14日,观察3组小鼠原位肿瘤生长情况及肺部转移情况。结果canstatin在转染重组载体的人淋巴细胞中表达并分泌至培养上清中。重组载体组小鼠肿瘤体积1.49cm3±0.18cm3明显小于空载体组(2.44cm3±0.19cm3)和NS组(2.53cm3±0.18cm3)(P=0.000)。重组载体组、空载体组和NS组的肺转移结节数分别为3.40±1.14、7.60±2.61和7.60±2.41,重组载体组明显少于后两组(P=0.013)。结论canstatin基因重组载体能在人淋巴细胞中表达并分泌至细胞外。canstatin可明显抑制小鼠Lewis肺癌移植瘤的生长与转移。
BACKGROUND AND OBJECTIVES In recent years, molecular targeted therapy for lung cancer has made significant progress. Targeted anti-angiogenic molecules have attracted widespread attention. The purpose of this study was to investigate the inhibitory effect of human lymphocyte culture supernatant transfected with human canstatin gene recombinant expression vector on the growth and metastasis of animal tumors. METHODS: Canstatin recombinant expression vector and empty vector were transfected into human lymphocytes by electroporation. G418 selection was used to obtain transgenic cell clones. The expression of canstatin protein in culture supernatant was detected by SDS-PAGE electrophoresis. Lewis lung cancer cells were inoculated subcutaneously in C57BL mice. After tumorigenesis, 30 mice were randomly divided into 3 groups. The supernatants of the lymphocyte culture supernatant transfected with the recombinant vector, the culture supernatant of the lymphocyte transfected with the empty vector, and the physiological saline were administered respectively. NS) 0.2 mL each was injected subcutaneously in the groin, once a day for 14 consecutive days, and the orthotopic tumor growth and lung metastasis of the three groups were observed. As a result, canstatin was expressed in human lymphocytes transfected with the recombinant vector and secreted into the culture supernatant. The tumor volume of the mice in the recombinant vehicle group was 1.49 cm3 ± 0.18 cm3, which was significantly smaller than that of the empty vehicle group (2.44 cm3 ± 0.19 cm3) and the NS group (2.53 cm3 ± 0.18 cm3) (P = 0.000). The numbers of pulmonary metastatic nodules in the recombinant vector group, empty vector group, and NS group were 3.40±1.14, 7.60±2.61, and 7.60±2.41, respectively. The recombinant carrier group was significantly less than the latter two groups (P=0.013). Conclusion The canstatin gene recombinant vector can be expressed in human lymphocytes and secreted extracellularly. Canstatin significantly inhibited the growth and metastasis of Lewis lung cancer in mice.