Enhancing Effect Of Tanshin On Suicide Gene Bystander Effect And The Mechanism Study

Gene therapy plays an important role in clinic coming from molecular biology technique. It could make effect with the translation outcome by taking exterior gene into normal or sick cells. During these years, it has made great development in gene therapeutic technique, which could be efficient therapy for tumor. It is great obstruction to because of lack of available measures for that lower level expression in the situation, introduced suicide gene and designed the suicide mechanism of the diseased cell, it is an important practise value for the anticancer.At present, HSV-TK is the most used, which is transfixed in tumour cells, cooperating GCV therapy , the expression of gene will turn GCV into GCV-TP , then kill tumour cells , but normal cells are not hurt .This use of therapy not only educe the death of cells transfecteded suicide gene , but also kill adjacent divided cells not transfecteded suicide gene by stander effect, and extend the effect of killing .It is important for the therapy of cancer .GJ is almost existed all of animals , dispose of no differentiate cells , eg. Skeletal muscle cells , red blood cells and circulate lympH cells . GJ has the passages , which are placed between cells , may pass small molecule matter by no choice , but not allowed pass large molecule and nucleic acid molecule .The expression product of GCV by direct touch of cells ,get in tk" cells .however , the pHospHoric acid product of GCV don' t get by a cell membrane .Thus , it is inferred that GJ of cells are thoroughfare of the pHospHoric acid product of GCV. The GJ of tumor pratial stander effect is tested by more experiments.Objective: To construct a new cell stain by transfect tk gene to H22 cell stain and name it H22/tk. Then use it for treating tumor in vivo and observethe curative effect of treatment by combining tan shin and GCV. And study the mechanism by immunohistochemical and TUNEL method.Through examination of the rat' s having tan shin serum and the CX-43 H22 cells, the mechanism is furthermore studied. Provide requirement for gene therapy of tumor by combining traditional Chinese and western medicine.Methods: Based on PT67/tk cell stain has been construction by DNA recombinant technique, then determining vival titer by infecting NIH 3T3 cells. H22 was infected with the recombinant retrovirus. The positive clones were obtained after G418 selection and named H22/tk. The morpHological changes after GCV treatment were observed with microscope. The liver tumor model on the KM mice were made in vivo by inoculate the cell mix which comprise 80% H22 and 20% H22/tk. Give a treatment by injecting pro-drug GCV, and combined with tan shin. Observe the weight of tumor after 16 days and histopathological expression of the tumors were observed in every group. Observe the expression of CX-43 of cells by immunohistochemical technic. Using cell Chemistry method and streamline cell skill to examine the CX-43 of cells, which are acted by tanshin.Conclusion: In vivo, transfer of HSV-tk gene plus GCV combined with tan shin have synergistic antitumor effects, and the mechanism of which relate with the promotion of stander effect and the expression of CX-43. In vitro.the expression of CX-43 is a higher level using the cell Chemistry method and streamline cell skill. The combined therapy might have therapeutic potentials for human cancer.