The Experimental Study Of Gene Therapy In Hepatcellular Carcinoma By Human Transfection Tumor Infiltrating Lymphocytes Transfected With TNF-a Gene

Object: In order to study the value of gene therapy in hepatocelluar carcinoma, transgene tumor infiltrating lymphocytes (transgene TIL) was constructed by transfected with tumor necrosis factor-α (TNF-α) gene into human TIL. The ant itumor activity of transgene TI[L was studied in vitro, and was applied in the treatment of the model of carcinoma-bearing nude mice. Methods : Gene transfer technique was used to transfer TNF-α gene into TIL which isolated from human hepatocellular carcinoma tissue, to construct a new type of antitumor effect cell-transgene TIL.The biological characters of transgene TIL was studed. The expression of aim gene was detected by RT-PCR, and the secretion of TNF-α was detected with TNF-α detected kit. The cytotoxic activity of transgene TIL was detected by MTT method. Human hepatoma cell line BEL7404 was injected into nude mice to establish the planted model of hepatocellular carcinoma, which was treated by transgene TIL. TIL was used in control group. 0 Results: The proliferation activity of transgene TIL was similar to TIL. Both of them had alike curve of cell proliferation. The cell surface phenotype of transgene TIL did not have obvious changed, the level of CD8+ was a little higher(60%), and transgene IlL still maintained higher stability. The- higher secretion of TNF-α was most significant change in transgene TIL, its expression dose was 370pg/105×105 cells/24h , but TNF-α secretion of normal TIL was negative. The cytotoxic activity of transgene TIL against human hepatoma cell line BEL7404 was depended on effect-target cell ration and action time. Using transgene TIL to treat the model of carcinoma-bearing nude mice, the significant therapeutic effect was achieved. Compared with normal TIL, transgene TI[L had stronger inhibit ion effect to generation rate, growth speed and transmigration of tumor. The antitumor activity of transgene IlL was showed up gradually with the passage of time and the change of acting environment. When transgene TIL was injected in the situs of tumor, a better antitumor effect was achieved . In addition different situs was injection had some effect, but its effects was lower than those injection in the situs of tumor. Conclusion: Transgene TIL is a kind of effective gene therapy in hepatocellular carcinoma. It is suggested that transgeng TIL may play a role in the treatment of patients with hepatoeelluar carcinoma.