Gene therapy of gastrointestinal cancer using adenoviral vector

探讨腺病毒用于胃肠道肿瘤基因治疗的可能性。首先，将具有插入的腺病毒（AxlacZ）的lacZ基因用作报告基因用于体外和体内感染实验。将来自食管癌、胃癌和大肠癌的培养细胞株置于体外与AxlacZ接触，并使用X-gal染色检查腺病毒感染性。在细胞株之间观察到感染性的相当大的差异，从约20%变化到接近100%。接下来，将细胞系皮下植入裸鼠，将AxlacZ直接注射到肿瘤中，几天后切除肿瘤。然后用X-gal染色，确认腺病毒感染。此外，使用BALB/c小鼠和来自它们的大肠癌细胞系Colon 26研究了腺病毒的细胞免疫。首先将AxlacZ注入小鼠腹腔，随后将结肠26感染 ...更多信息 植入小鼠体内。与未给予AxlacZ预处理时相比，肿瘤生长受到抑制，表明细胞免疫的建立。接下来，制备表达野生型p53的重组腺病毒（Axp 53），并用于在体外和体内研究食管癌细胞系中的抗肿瘤效果。在体外用Axp 53感染细胞系，并在MTT测定中检查细胞存活。根据细胞系，观察到约20%至90%的变化。在体内研究中，将细胞系皮下植入裸鼠中，并直接注射AxlacZ或Axp 53。用AxlacZ观察到细胞增殖的抑制，但Axp 53被证明甚至更有效。在Axp 53注射区域，通过免疫染色证实p53蛋白表达。Axp 53与抗癌剂的组合也在体外和体内进行了研究，但仅获得了累加效应。少

The possibility of using adenoviruses for gene therapy of gastrointestinal cancer was investigated. First, a lacZ gene with an inserted adenovirus (AxlacZ) was used as a reporter gene for in vitro and in vivo infection experiments. Cultured cell strains from esophageal, stomach, and large intestinal cancers were placed in contact with AxlacZ in vitro, and the adenovirus infectivity was examined using X-gal stain. A considerable difference in infectivity was seen between the cell strains, varying from about 20% to nearly 100%. Next, the cell line was implanted subcutaneously in nude mice, AxlacZ was injected directly into the tumor, and several days later the tumor was excised. It was then stained with X-gal, and the adenovirus infection was confirmed. In addition, cellular immunity from the adenovirus was investigated using BALB/c mice and large intestinal cancer cell line colon 26 from them. AxlacZ was first administered into the mouse abdominal cavity, and afterward colon 26 infected … More with AxlacZ was implanted into the mice. Tumor growth was suppressed compared to when no AxlacZ pre-treatment was given, suggesting establishment of cellular immunity.Next, recombinant adenovirus (Axp53) expressing wild-type p53 was prepared, and used to investigated the antitumoral effect in the esophageal cancer cell line in vitro and in vivo. The cell line was infected with Axp53 in vitro and cell survival was examined in an MTT assay. A variation of about 20% to 90% was seen, depending on the cell line. In the in vivo investigation, the cell line was implanted subcutancously in nude mice, and AxlacZ or Axp53 was injected directly. Suppression of cell proliferation was seen with AxlacZ,but Axp53 proved to be even more effective. In the region of the Axp53 injection p53 protein expression was confirmed by immunostaining. The combination of Axp53 with an anticancer agent was also investigated both in vitro and in vivo, but only an additive effect was obtained.In light of the foregoing, p53 recombinant adenovirus is seen to be an effective gene therapy for gastrointestinal cancer. Less