Gene therapy of gastrointestinal cancer using adenoviral vector

探讨了利用腺病毒进行胃肠道肿瘤基因治疗的可能性。首先，利用插入腺病毒（AxlacZ）的lacZ基因作为报告基因进行体外和体内感染实验。将体外培养的食管癌、胃癌和大肠癌细胞株与AxlacZ接触，用X-gal染色法检测腺病毒的感染性。不同细胞株之间的传染性有相当大的差异，从约20%到近100%不等。接下来，裸鼠皮下植入细胞系，AxlacZ直接注射到肿瘤中，几天后切除肿瘤。用X-gal染色，证实腺病毒感染。此外，用BALB/c小鼠和大肠结肠癌细胞系结肠癌26研究了腺病毒对细胞的免疫作用。首先将AxlacZ注入小鼠腹腔，然后将AxlacZ植入小鼠结肠。与未给予AxlacZ预处理相比，肿瘤生长受到抑制，提示细胞免疫的建立。下一步，制备表达野生型p53的重组腺病毒（Axp53），在体外和体内研究其对食管癌细胞系的抗肿瘤作用。用Axp53体外感染细胞系，用MTT法检测细胞存活。根据细胞系的不同，变异率约为20%至90%。在体内研究中，裸鼠皮下植入细胞系，并直接注射AxlacZ或Axp53。AxlacZ可以抑制细胞增殖，但Axp53被证明更有效。免疫染色证实在Axp53注射区p53蛋白的表达。体外和体内也研究了Axp53与抗癌药物的联合作用，但只获得了加性效应。鉴于上述，p53重组腺病毒被认为是一种有效的基因治疗胃肠癌。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