摘要
目的探讨直肠癌组织中脑特异性血管生长抑制因子1(BAI1)基因的表达情况及BAI1对肿瘤血管生成和肿瘤生长的作用。方法提取20例直肠癌患者(II期6例,III期14例)手术切除的直肠癌、无转移淋巴结、有转移淋巴结组织和直肠腺癌细胞株CC95总RNA,应用逆转录聚合酶链反应方法检测BAI1的表达。将背部皮下接种CC95细胞(1×105个)的BALB/C裸鼠随机分成3组,每组10只,分别在接种后第1、3、5、7、9天于荷瘤部位注射RPMI1640、腺病毒载体(Ad-LacZ)或基因重组BAI1表达腺病毒载体(Ad-hBAI1),每次0.2ml,载体含量2μg。每组中5只裸鼠于接种后第15天断颈处死,取肿瘤组织行抗Ⅷ因子抗体染色,观察微血管密度(MVD);另5只接种后每天观察肿瘤体积的变化,共观察21d。结果20例患者中,7例(临床II期6例、III期1例)肿瘤组织BAI1表达呈阳性,13例无表达;无转移淋巴结均呈阳性表达,有转移淋巴结均无表达。CC95细胞均无BAI1表达。在BALB/C裸鼠肿瘤移植实验中,Ad-hBAI1组肿瘤组织内血管增生受到明显抑制,MVD(6.2±2.3)明显低于RPMI1640组(16.7±1.9)和Ad-LacZ组(13.4±4.3)(均为P<0.05);肿瘤生长速度明显低于RPMI1640组和Ad-LacZ组(均为P<0.01)。结论BAI1在人直肠癌及转移的淋巴结组织中表达降低;局部注射Ad-hBAI1表达载体可抑制肿瘤血管生成并抑制肿瘤的生长。
Objective To explore the expression of brain-specific angiogenesis inhibitor 1 (BAI1) gene in rectal cancer tissues and its effect on growth and angiogenesis of the tumor. Methods Tissue samples of rectal cancer, non-metastatic lymph nodes, and metastatic lymph nodes were collected from 20 patients with rectal cancer (6 with stage II and 14 with stage III). The total RNA was extracted from rectal adenocarcinoma cell line (CC95) and the tissues from the patients, and reversely transcribed to cDNA using RT-PCR to detect the expression of BAI1 gene. Thirty BALB/C nude mice inoculated with CC95 cells ( 1×105) on the back were randomly divided into 3 groups (10 in each). RPMI1640, adenoviral vector (Ad-LacZ), and recombinant adenoviral vector expressing human BAI1 gene (Ad-hBAI1) were injected locally to the tumors in the mice of the three groups respectively on day 1, 3, 5, 7, and 9 (2 ml/time, containing 2 μg vectors). On the fifth day after the inoculation, 5 mice of each group were killed; the tumor tissues were excised from the mice and stained with anti-factor VIII antibodies to determine the microvessel density (MVD). The other 5 mice of each group were observed for 21 days to record the change of the tumor volume. Results The BAI1 gene was expressed in 7 (6 with stage II and 1 with stage III) of the 20 rectal cancer tissue samples and all the non-metastatic lymph nodes, but in none of the metastatic lymph nodes and CC95 cells. The MVD in the Ad-hBAI1 group (6.2±2.3) was significantly lower than that in the RPMI1640 (16.7±1.9) and Ad-LacZ (13.4±4.3) groups, showing that Ad-hBAI1 markedly inhibited the angiogenesis of the tumor (both P<0.05). The growth rate of the tumor in the Ad-hBAI1 group was also significantly lower than that in the RPMI1640 and Ad-LacZ groups (both P<0.01). Conclusions BAI1 is lowly expressed in human rectal cancer tissues and the metastatic lymph nodes. Local injection of Ad-BAI1 recombinant vector can inhibit the angiogenesis and growth of the tumor.
出处
《中国医药生物技术》
CSCD
2006年第1期28-31,共4页
Chinese Medicinal Biotechnology
基金
黑龙江省重点攻关课题(G00C190401)
