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目的:探讨携带多基因(抑癌蛋白:p53,免疫共刺激分子:B7- 1,粒细胞巨噬细胞集落刺激因子:Granulocyte-Macrophage Colony- Stimulating Factor,GM-CSF)的复制缺陷型腺病毒(BB-102)对喉鳞癌原代细胞中的表达效率和对其生长的抑制作用。方法:构建多基因联合(p53,GM-CSF,B7-1)重组的复制缺陷腺病毒BB-102, 在体外感染两株喉鳞癌细胞系(LSC-1及L-17),采用免疫组化, 刘氏细胞术,ELISA等检测目的基因编码蛋白效率,用MTT法观察其对喉癌细胞生长抑制作用。结果:三种目的基因在喉癌细胞中均有不同程度的表达:B7-1从平均1．34％提高到57％;GM-CSF在转染后24小时即开始升高,持续两天后低水平维持在一周时间;p53蛋白表达明显,但当50 pfu／细胞的相同浓度转染时,其表达效率没有Ad-p53高。BB-102和Ad-p53分别对肿瘤细胞生长产生抑制作用, 两者没有显著差异(P>0．05)。结论:BB-102感染喉癌细胞,目的基因都能有效表达,喉癌原代细胞生长受到明显抑制;感染滴度相同时,单基因腺病毒Ad-p53感染的肿瘤细胞p53蛋白表达丰度比BB- 102高。图5表1参9
OBJECTIVE: To investigate the effect of replication-defective adenovirus carrying multiple genes (tumor suppressor protein: p53, immune costimulatory molecule: B7-1, Granulocyte-Macrophage Colony-Stimulating Factor (GM-CSF) BB-102) on the expression of primary laryngeal squamous cell carcinoma cells and their growth inhibition. Methods: The replication-deficient adenovirus BB-102 with multiple genes (p53, GM-CSF, B7-1) was constructed and in vitro infected with two LSCC-1 and L-17 cells. Liu, cytometry, ELISA and other detection of the target gene encoding protein efficiency, MTT assay was used to inhibit the growth of laryngeal carcinoma cells. Results: The expression of all three target genes in laryngeal carcinoma cells was different: B7-1 increased from an average of 1.34% to 57%; GM-CSF began to rise 24 hours after transfection, and remained low for two days The levels were maintained for a week; the expression of p53 protein was significant, but when the same concentration of 50 pfu / cell was transfected, its expression efficiency was not high in Ad-p53. BB-102 and Ad-p53 inhibited the growth of tumor cells, respectively, with no significant difference (P> 0.05). CONCLUSIONS: The target gene of BB-102 is highly expressed in laryngeal carcinoma cells and the growth of laryngeal carcinoma primary cells is significantly inhibited. When the titer of infection is the same, the expression abundance ratio of p53 protein in single-gene adenovirus Ad-p53-infected tumor cells BB-102 high. Figure 5 Table 1 参 9