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目的探讨脑组织特异性血管生成抑制因子1(BAI1)对胶质母细胞瘤的治疗作用及其作用机制。方法采用COS-TPC法进行BAI1-腺病毒载体的构建,病毒重组子的成功构建和对肿瘤细胞的转染通过RT-PCR来验证。采用立体定向的方法将胶质母细胞瘤细胞U87MG接种于裸鼠脑内,待肿瘤形成后向瘤内注射病毒重组子AdeBAI1(AdeBAI1组)或AdeLacZ(Mock)(AdeMock组),两组均为6只裸鼠。观察裸鼠的存活时间。用AdeBAI1或AdeMock转染3个胶质母细胞瘤细胞系SW1783、U87MG和U373MG,48h后收集细胞,用MTT方法进行活细胞计数。用Trizol试剂提取总RNA。用RT-PCR方法研究BAI1及其他血管生成相关因子的mRNA的表达。结果BAI1mRNA的表达仅见于AdeBAI1转染的细胞中。颅内胶质母细胞瘤治疗结果显示,AdeBAI1组平均生成期为(26·0±4·6)d,AdeMock治疗组为(17·3±2·3)d,两组比较P<0·05。AdeBAI1组转染后细胞计数为(2·12±0·18)×105、AdeMock组为(4·23±0·18)×105,两组比较P<0·05。提示胶质母细胞瘤细胞的增殖可被BAI1抑制。AdeBAI1转染后血管生成相关因子Angiostatin和VEGF的表达降低,而VEGF-B和TSP1的表达升高。结论肿瘤内注射AdeBAI1可以抑制胶质母细胞瘤的生长。BAI1的表达升高可以影响其他血管生成相关因子的表达。BAI1的表达升高可以抑制肿瘤细胞的增殖作用。BAI1的抗肿瘤作用可能来自于抑制血管生成以及抑制肿瘤细胞的增殖两个方面。
Objective To investigate the therapeutic effect and mechanism of brain tissue-specific angiogenesis inhibitor 1 (BAI1) on glioblastoma. Methods COS-TPC method was used to construct BAI1-adenovirus vector. The successful construction of recombinant virus and the transfection of tumor cells were confirmed by RT-PCR. The glioblastoma U87MG cells were inoculated into the brains of nude mice by stereotactic method. After the tumors were formed, the virus-injected recombinant AdeBAI1 (AdeBAI1) or AdeLacZ (Mock) (AdeMock) 6 nude mice. Observe the survival time of nude mice. Three glioblastoma cell lines, SW1783, U87MG and U373MG, were transfected with AdeBAI1 or AdeMock. Cells were harvested 48 hours later and viable cells were counted by the MTT method. Total RNA was extracted with Trizol reagent. The mRNA expression of BAI1 and other angiogenesis-related factors was studied by RT-PCR. Results BAI1 mRNA expression was found only in AdeBAI1 transfected cells. The results of intracranial glioblastoma treatment showed that the average age of AdeBAI1 group was (26.0 ± 4.6) days and that of AdeMock group was (17.3 ± 2.3) days, P <0 · 05. The number of cells in the AdeBAI1 group was (2.12 ± 0.18) × 105 after transfection, (4.23 ± 0.18) × 105 in the AdeMock group, P <0.05 compared with the two groups. It suggests that the proliferation of glioblastoma cells can be inhibited by BAI1. The expression of Angiostatin and VEGF decreased after AdeBAI1 transfection, while the expression of VEGF-B and TSP1 increased. Conclusion Intradermal injection of AdeBAI1 can inhibit the growth of glioblastoma. Increased BAI1 expression can affect the expression of other angiogenic factors. BAI1 expression can inhibit the proliferation of tumor cells. The antitumor effect of BAI1 may come from the two aspects of inhibiting angiogenesis and inhibiting the proliferation of tumor cells.