摘要
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 objective In recent years, many progresses have been made in molecular target therapy for lung cancer, in which anti-angiogenic target therapy is a hot spot drawing researchers＇ attention. The aim of this study is to explore the expression of canstatin gene transfected into human lymphocytes and its inhibitory effect on growth and metastasis of Lewis lung carcinoma. Methods The eukaryotic expression vector of pCMV-Script and the recombinant pCMV-Script/Canstatin vector were separately transfected into lymphocytes by electroporation. The expression of canstatin protein in supernatant of lymphocytes was examined by SDS-PAGE assay. Furthermore, Lewis lung carcinoma cells were subcutaneously inoculated to C57BL mice to make animal model of tumor. When the transplanted tumors on the mice developed to 1 cm3 , the 30 mice were randomized into 3 groups, which were injected with 0. 2 mL supernatant of lymphocytes transfected with recombinant vector or naked vector, or 0.2 mL NS respectively. After the treatment for 14 days, the size and pathological section of subcutaneous tumors were observed, and the number of pulmonary metastatic node was calculated. Results Canstatin protein was found in supernatant of the lymphocytes in the recombinant vector group by SDS-PAGE assay. After the treatment, the tumor size in the recombinant vector group （1.49 cm^3 ±0. 18 cm^3） was significantly smaller than that in the naked vector group （2. 44 cm^3±0.19 cm^3 ） and NS group （2.53 cm^3±0. 18 cm^3 ） （P=0. 000）. The numbers of pulmonary metastatic node were 3.40±1.14, 7.60±2.61 and 7.60±2.41 in the recombinant vector group, naked vector group and NS group respectively （recombinant vector group vs the other two groups, P=0. 013）. Conclusion The pCMV-Script/ Canstatin vector can express canstatin protein in human lymphocytes. Canstatin has strongly inhibitory effect on growth and metastasis of mouse Lewis lung carcinoma.
出处
《中国肺癌杂志》
CAS
2006年第3期245-249,共5页
Chinese Journal of Lung Cancer
基金
第三军医大学校管课题基金(2005D125)资助~~
